Guanidinoacetic Acid (GAA) 'Superior' to Creatine in Terms of Bioenergetic & Health Effects on Brain, Muscle & More?

Not so fast! Just because it may have shuttled more creatine into the muscle GAA does not have to be able to build more strength or size.

It has been a while since the last creatine article was published. Luckily, the publication of the latest paper from Sergej M. Ostojic lab at the University of Novi Sad (Ostojic. 2016b) will change that... as soon as I've analyzed the study and written about it below that is ;-)

First things first, we are dealing with a randomized, double-blind, cross-over trial that was financed by a government grant - not non-existing (as of now) supplement companies who are pimping guanidinoacetic acid (GAA) as a "superior new form of creatine".

You can learn more about creatine at the SuppVersity

Creatine Doubles 'Ur GainZ!

Creatine Loading = Unnecessary

Creatine Pre or After Workouts?

1st Benefits of Creatine-HCL

The Real Bioavai-lability of Crea

Build 'Ur Own Buffered Creatine

Now, technically speaking, this claim would be bogus, anyways, GAA is, after all, an intermediate product formed enzymatically in the liver, pancreas, and kidney in the synthesis of creatine and not "a creatine" - "superior", or not.

The formation of increasing amounts of S-adenosylhomocysteine (SAH) ad subsequently homocysteine (HCY, not shown in the figure, which focuses on the synthesis of creatine) which is potentially bad for your heart- and brain will have to be controlled and means to blunt it will have to be found in future studies (da Silva. 2009).

In the study, of which I want to highlight that the number of subjects limits the significance of its results significantly (consider everything as preliminary evidence), five healthy young men (age 24.3 ± 3.1 years; body mass index 24.9 ± 1.9 kg/m²) volunteered to help the scientists find out, whether 4-week supplementation with 3g / day guanidinoacetic acid (GAA) would be superior to 3.4 g of creatine in facilitating creatine levels and thus have potential performance and health enhancing effects that go beyond those of the 'real thing' (creatine).

Figure 2: Percentage change of tissue creatine levels 0 vs. 4 weeks. GAA denotes guanidinoacetic acid. Values are presented as means, with error bars represent the standard error of the means (SEM). P between trials (Ostojic. 2016b).

As you can see in Figure 2, "GAA (3.0 g/day) resulted in the more powerful rise (up to 16.2%) in tissue creatine levels in vastus medialis muscle, middle cerebellar peduncle, and paracentral gray matter, as compared to creatine" (P < 0.05 | Ostojic 2016).

Another (add. cf. Fig 1) explanation for GAA's ill effect on homocysteine is that its conversion to creatine requires methyl donors that can then no longer be used to "recycle" cysteine. We'll have to see if supplementing met-don alone will solve the problem, though.

How does GAA work? GAA yields creatine mainly in the liver (also pancreas and kidney), w/ prev. studies (by the same group) confirming the creatine-boosting potential of dietary GAA in serum and skeletal muscle of healthy men and women (Ostojic et al. 2014; Ostojic et al. 2016a). As Ostojic et al. explain, "GAA is absorbed from the intestinal tract, enters the circulation and is methylated to creatine by guanidinoacetate N-methyltransferase (GAMT); substrates of this enzyme are GAA and S-adenosyl methionine, whereas its two products are creatine and S-adenosyl homocysteine. Usually considered as a less adequate source of cellular energy" (Ostojic. 2016b).

Next to the formation of homocysteine during the conversion process of GAA to creatine, the fact that this process requires additional methyl donors could be another reason why Ostojic et al. (2014) found that Hcy started to pile up in the blood of their GAA study (see Figure 3). Similar neg. effects on Hcy have been reported before; among others by Stead, et al.  (2001). And it seems only logical: If the methyl-donors are used for the GAA - creatine conversion they are no longer available to recycle homocysteine so that the latter is increasing progressively. This is bad news, since a high serum HCy concentration is an independent risk for cardiovascular disease and Alzheimer’s disease, dementia and cognitive impairment (Morris. 2003; Ganguly. 2015) - the causal nature of the epidemiologically observed link has yet been put into question, repeatedly, but until GAA receives the label "clinically proven safety" I cannot recommend using it as a replacement for creatine.

Ostojic et al. interpret (they didn't test and thus prove that their albeit logical interpretation is adequate) that these results signify that GAA would be "a preferred alternative to creatine for improved bioenergetics in energy-demanding tissue" (my emphasis in Ostojic. 2016). Needless to say that this will require

• 

  Figure 2: Fasting total plasma homocysteine concentration at baseline and at 1, 2, 4, and 6 weeks of supplementation (Week 1, Week 2, Week 4 and Week 6, respectively) in subjects receiving placebo or guanidinoacetic acid (GAA) for 6 weeks at 1.2, 2.4 or 4.8 g/day, respectively (Ostojic. 2014).

  experimental confirmation in studies testing the actual effects on cellular bioenergetics and health effects in energy-demanding tissues such as muscle and brain, 
• further investigations of the potential (health-)relevance of the subsequent increase in methylation reactions that occur during the formation of creatine from GAA in the human body (more specifically, the question "Is the sign. increase in homocysteine levels a health problem for GAA users?" will have to be answered - esp. in the long-term; after all, Ostojic's 2014 study indicated that these start to raise with time / Figure 3), and
• long(er) term studies and studies comparing different dosages of GAA and creatine; after all, the former is supposed to work, because it offers a greater variety of uptake channels - an advantage that may be easy to compensate by (a) administering more creatine (3.4g is not much) and/or for a longer time (4 weeks is short)
• independent confirmation of the experimental results from another lab to exclude a potential bias of the researchers who have been working on the issue for years, now.

Not a single one of these studies has yet been conducted. So that we are left, for the time being, with this short-term study with a very 'limited' (you could also say 'hardly sufficient') number of subjects.

If you haven't read it, yet I suggest you read up on my previous article about a study in Elite Footballers, where high doses of creatine actually resulted in inferior effects on body composition than lower doses. Quite an interesting result in view of the "more helps more"-mentality that's prevalent in the fitness community | Read the SV Classic from October 2015!

Bottom line: Why should GAA even be better than creatine? While Ostojic et al. still have a lot of work to do to actually prove that GAA is (a) safe and (b) "better" than creatine (namely in studies with relevant study outcomes, like performance or lean mass increases, for example) they do already have a hypothesis why it could be:

"This perhaps happens due to preferable uptake of GAA by target tissues via various mechanisms theoretically available for GAA transport, as compared to somewhat limited transport capacity of creatine, and/or complete (or near to complete) tissue methylation of GAA to creatine. While Cr is mainly transported via specific transporter (SLC6A8; also used for GAA transport), dietary GAA could be imported through additional delivery channels (SLC6A6, GAT2, passive diffusion) at least in the brain, and become readily available for tissue methylation to creatine by GAMT" (Ostojiv. 2016b).

In other words: GAA is incorporated into muscle and especially brain by more pathways than creatine and may thus be able to increase the creatine stores in said organs faster and to a greater extent than creatine and that's why it could be the 'better creatine' - one that lacks long-term safety data and convincing evidence that the advantage is practically relevant, though. I guess I don't have to tell you, then, that it is not (yet?) "SuppVersity suggested" | Comment!

References:

• da Silva, Robin P., et al. "Creatine synthesis: hepatic metabolism of guanidinoacetate and creatine in the rat in vitro and in vivo." American Journal of Physiology-Endocrinology and Metabolism 296.2 (2009): E256-E261.
• Ganguly, Paul, and Sreyoshi Fatima Alam. "Role of homocysteine in the development of cardiovascular disease." Nutrition journal 14.1 (2015): 1.
• Morris, Martha Savaria. "Homocysteine and Alzheimer's disease." The Lancet Neurology 2.7 (2003): 425-428.
• Ostojic, Sergej M., et al. "Dose–response effects of oral guanidinoacetic acid on serum creatine, homocysteine and B vitamins levels." European journal of nutrition 53.8 (2014): 1637-1643.
• Ostojic, Sergej M., Patrik Drid, and Jelena Ostojic. "Guanidinoacetic acid increases skeletal muscle creatine stores in healthy men." Nutrition 32.6 (2016a): 723-724.
• Ostojic, Sergej M., et al. "Guanidinoacetic acid vs. creatine for improved brain and muscle creatine levels: a superiority pilot trial in healthy men." Applied Physiology, Nutrition, and Metabolism (2016).
• Stead, Lori M., et al. "Methylation demand and homocysteine metabolism: effects of dietary provision of creatine and guanidinoacetate." American Journal of Physiology-Endocrinology And Metabolism 281.5 (2001): E1095-E1100.

Choline Deficiency, Its Consequences and How You Fix It | Part 2 of the "Common Nutrient Deficiencies, Their Health Consequences and How You Can Fix Them" Series

Don't you tell me there was no fancy cooking with the choline source #1, i.e. egg yolk! What about egg yolk on ricotta cauliflower ravioli filling, for example, | get the recipe @ tastespotting.com

If you've read the last installment of this series, you can hardly be surprised that today, we are about to talk or rather I am about to write about choline. Choline is, as Wikipedia informs us, a water-soluble essential nutrient. Why choline is not officially called a "vitamin" is beyond me. "Experts" will still group it within the B-complex vitamins, anyway.

In my analyses in the series on nutrients other than carbohydrates that influence glucose control, you've read that the former, the "other", "real" B-vitamins are - in my humble opinion - totally overrated. Choline, which can come in various forms of quaternary ammonium salts all of which contain the characteristic N,N,N-trimethylethanolammonium cation, on the other hand, it probably the most underrated micronutrient - not only for glucose control, by the way.

You can learn more about choline at the SuppVersity

Huge GH Spikes from GPC!?

Choline & Classic Bodybuilding

Improved Lipid Profile W/ Eggs

Caffeine, Choline, L-Car = Fat Loss

Choline Maximizes Carnitine Storage

Choline as a Nootropic?

One of the reasons choline has been depreciated is probably that it occurs in all the good foods of which scientists have been (falsely) telling you that you must not eat them for decades.

Figure 1: Eggs, or rather their fatty yolks, are a very good source of choline. One too few people appreciate for its nutritional density and too many people fear for its allegedly bad cholesterol content | learn why this is bullocks

Voilà, exhibit A (see Figure 1), an egg. The "bad" yolk of a single egg contains enough choline to "get you through the day" - whether that's also enough choline for optimal health is a question we are about to tackle later in today's installment of the Common Nutrient Deficiencies, Their Health Consequences and How You Can Fix Them" Series (you can browse through past and previous issues, here, if you have an RSS compatible web-browser).

The fact that generations of bodybuilders have been throwing away the yolks only to supplement with even more choline than they'd have gotten from their eggs does yet certainly tell you something about the importance of this nutrient.

But I am digressing, here. What we actually wanted to talk about is after all "normal" people, not bodybuilders. Normal people like the participants of the 2005 National Health and Nutrition Examination Survey (NHANES) among which only 2% of postmenopausal women consumed the recommended amount of for choline, and in general "mean choline intakes for older children, men, women and pregnant women are far below the Adequate Intake established by the IOM" (Zeisel. 1991).

When people don't get enough choline in their diets, and the amount of choline in their system declines, will eventually lose their ability to methylate homocysteine to methionine. The result, their plasma levels of homocysteine increase and heart disease, dementia, and even cancer are lurking right around the corner.

Why is high plasma homocysteine bad? In spite of the fact that it is still questionable, whether homocysteine is causally involved in the etiology of heart disease (Brattström. 2000), the present evidence leaves little doubt that high homocysteine levels are a marker of increased risks for several chronic diseases and conditions including cardiovascular disease, cancer, cognitive decline and bone fractures (HSC. 2002; Wu. 2002; Seshadri. 2002; van Meurs. 2004).

Even in the absence of vitamin B6, and B12, of which everybody is talking these days, choline and its cousin betaine have the ability to lower homocysteine levels (Chiuve. 2007). In this context you should also keep in mind that (a) homocysteine levels and the development of the triage we call "metabolic syndrome" (obesity, diabetes, high cholesterol) and that (b) choline is "lost" (oxidized) during the methylation process -- the choline requirements of the average, meanwhile at least chubby US citizen are thus naturally higher than the RDA scientists have determined in a day and age, when a BMI of >30 was still the exception.

Figure 2: The maintenance of healthy homocysteine levels is only one of choline's many important effects.

Aside from being involved in the clearance of homocysteine, choline is also, and maybe even more importantly, used in the synthesis of the constructional components of your body's cell membranes and much more (see Figure 2, as well):

• Choline figures in cell signaling, where the choline-containing phospholipids, phosphatidylcholine, and sphingomyelin, are precursors for the intracellular messenger molecules, diacylglycerol and ceramide. And with the platelet activating factor (PAF, an activator, and mediator of many leukocyte functions, including platelet aggregation and degranulation, inflammation, and anaphylaxis) and sphingophosphorylcholine (regulate traffic around / across the cell membrane; Ramstedt. 2002), there are two additional choline metabolites with important systemic functions.
• The transmission of never impulses is another, certainly one of the most important functions of choline or rather acetylcholine, an important neurotransmitter involved in muscle control, memory, and many other functions. No wonder that low choline levels have been associated with the progression of Alzheimer's and non-Alzheimer's dementia (Babic. 1999; Seshadri. 2002)

Apropos beyond methylation - choline can make up for a lack of folate! The often decried lack of folate intake (just as a reminder the folate fortification program in the US made no one healthier - with one exception, maybe: The recommendations to increase folate intake for pregnant women/women of childbearing age has been relatively successful in preventing neural tube defects in infants) would be much less of an issue in the diabesity problem, if the average American consumed 2,200mg of choline per day. The same amount of choline that is which preserved the markers of cellular methylation and attenuated folate deficiency related DNA damage in a genetic subgroup of folate-compromised men in a 2010 study by Shin et al.

• The effects on lipid (fat) transport and metabolism are probably still totally underrated by both the public and certain parts of the medical establishment. The fat and cholesterol that's consumed in the diet are after all transported to the liver by lipoproteins (chylomicrons) which are built from phosphatidylcholine. If there is not enough choline to build these 'cholesterol shuttles' the fat and cholesterol will begin to accumulate in your liver. No wonder scientists have long discovered an intricate relationship between choline intakes, on the one hand, and the development of non-alcoholic fatty liver disease, on the other hand - in fact, more recent evidence suggests that millions of US citizens have been able to escape a non-alcoholic fatty liver only due to "good genes" that allow them to budget their insufficient choline intake better than others (Spencer. 2011).

In spite of the fact that a full-blown choline deficiency will impair your exercise performance, the provision of supplemental choline has not been shown to consistently produce meaningful increases in exercise performance (Penry. 2008). As a physical culturist, you should still keep in mind that intense exercise will increase your basal choline requirements of which the wise USDA says based on data of which the researchers say that its quality "varies widely across studies", with the most significant sources of potential sources being over- or underreporting of portion sizes and frequency of intake, omission of foods, and inaccuracies related to the use of food composition tables. Accordingly Yates et al. add for consideration:

Table 1: High choline foods for omnivores and vegetarians; choline content in milligrams / 100g | data adapted from nutritiondata.com

"Therefore, the values reported by nationwide surveys or studies that rely on self-report may be somewhat inaccurate and possibly biased. Food composition databases that are used to calculate nutrient intake from self-reported and observed intake data introduce errors due to random variability, genetic variation in the nutrient content, analytical errors, and missing or imputed data.

In general, when nutrient intakes for groups are estimated, the effect of errors in the composition data is probably considerably smaller than the effect of errors in the self-reported intake data (NRC, 1986). However, it is not known to what extent this is true for folate, biotin, pantothenic acid, or choline." (Yates. 1998)

That's bad because the first signs of liver damage in humans (beginning NAFLD if you will) occur after only 3 weeks on a choline deficient diet (Zeisel. 1991). In view of the fact that doses of safe therapeutic doses go into the 8-10 gram range and considering the fact that the total amount of choline in foods is not exactly exorbitantly high, it would appear prudent to follow the "more is better principle", when it comes to choline-rich foods (see Table 1 and bottom line for suggestions | Didn't find the food you were looking for? Try the official USDA Overview).

Don't go overboard on supplements! When it comes to supplementation, on the other hand, you better be careful not do overdo it. While choline is generally relatively benign, it can produce side effects that range from increased potassium and magnesium requirements over depressive-like symptoms (a general sluggishness) to vomiting and, in some people, acne-like skin rashes. Dosages in the below 3g per day range do yet generally appear to be tolerated very well. In the long term, the phospholipid-bound forms of choline can yet produce quite nasty depressive-like side-effects - another reason to prefer the cheap and effective citrates and tartrates if you are just "supplementing" your diet or looking for the metabolic effects, primarily.

Unless you are following a no fat diet, the use of dietary supplements to cover your baseline requirements of 500-600mg (I am deliberately not using the ostensibly "accurate" RDA, here) shouldn't  be necessary - at least if you eat your healthy eggs every day ;-)

Choline as a "metabolic activator" and diet aid!?

If you mimic the old-school body- builders and have your choline supps with meat, this will increase the retention & effects of carnitine.

If you remember my previous article on the relatively unknown "CCC-Stack" which consists of caffeine, carnitine, and... you guessed it, choline you will yet remember that doubling your intake with 500mg of choline tartrate or citrate had quite remarkable effects on the metabolism (learn more | for metabolic purposes I would prefer the citrate and tartrate forms over the expensive phospholipid bound version of choline, which may have its advantages, though, when it comes to brain-related effects).

Moreover, a recent study from the Zagazig and the Mansura University in Egypt suggests that choline alone, will produce quite impressive weight loss effects - even in the absence of the "fat liberator" caffeine and the "fat transporter" carnitine, i.e. the other "C"s in the CCC stack.

The study, Elsawy, Abdelrahman and Hamza conducted investigated the effects of choline supplementation on body mass reduction and leptin levels among female taekwondo and judo athletes in the pre-competition phase (Elsawy. 2014).

Figure 3: Relative changes in lipid oxidation, body fat (%), body mass and strength parameters in female athletes during contest prep with (choline) and without (control) 2x2g of supplemental choline in their meals (Elsawy. 2014)

Twenty-two female athletes (15 taekwondo and 7 judo athletes) were selected from different weight
categories and divided into two groups, according to weight. The players in the experimental group consumed their choline supplements (2x2.0g of choline bitartrate) daily with meals for one week.

I guess the results actually speak for themselves. As expected, the experiment revealed significant differences between pre- and post-competition measurements of free plasma choline, and urine choline levels. The significantly higher fat loss, on the other hand, is something even the researchers did not necessary expect to see. In view of the reduction in malondialdehyde (MDA = lipid oxidation) and the increase in carnitine retention and effects you've read about in a previous SuppVersity article ("Choline Maximizes Carnitine Retention + Effects" | read more) it is yet eventually not surprising that the choline group lost more body fat without suffering significant reductions in strength and lean mass.

Eat three of these or combine two of them: One whole eggs, 4oz of shrimp, or 5oz scallop, 5oz chicken, 5oz turkey

Eat four of these or combine four of them: 4oz cod, 1.5 cups of collard greens, 1.75 cups of Brussel sprouts, 1.75cups of broccoli, 5oz tuna, 5oz salmon, 6oz beef, 6 oz sardines

Eat five of these or combine one of each 2cups of swiss chard, 2cups of cauliflower, 2cups of asparagus, 2.25cups of spinach, 2cups green peas, 2.5 cups cabbage, 2 cups of Shiitake mushrooms

Other foods with choline, of which you'd yet probably have to eat too much to get to your SRI (SuppVersity recommended intake for adults) of 500-600mg are green beans and bok choy (5% of SRI per cup), summer squash, miso and tomatoes (1-2% of SRI per cup)

Bottom line: I am well aware that the fat loss effects, I deliberately mentioned last are probably the "sexiest" benefits choline has to offer. Compared to the conservation of liver, brain and heart hearth for which you "just" have to make sure that you consume enough of the foods in the list to the right, the ability of supplemental choline to promote fat loss and conserve lean mass and strength in "average Janes" (read more) and competitive athletes (Figure 3) is actually quite pathetic.

Speaking of "pathetic" the notion that eggs would be bad for you because they contain cholesterol is probably even more pathetic than the notion that being ripped was more attractive than being healthy. In view of the fact that the amount of highly bioavailable choline in a single egg can cover 20%-35% (depending on its size) of your daily choline demands, having an egg for breakfast is nothing you should consider, but rather something you should simply do! The increase in choline intake, the beneficial effects on your cholesterol particle and phospholipid profile and the increase in HDL-driven lipid reverse-transport are after all something you don't want to sacrifice on the altar or unwarranted prejudices and in blind obedience to dietary guidelines of which more and more researchers say that they "are not benefiting the public as a whole and may actually have negative nutritional implications." (Herron. 2004)

References:

• Babic, T. "The cholinergic hypothesis of Alzheimer’s disease: a review of progress." Journal of Neurology, Neurosurgery & Psychiatry 67.4 (1999): 558-558.
• Brattström, Lars, and David EL Wilcken. "Homocysteine and cardiovascular disease: cause or effect?." The American journal of clinical nutrition 72.2 (2000): 315-323.
• Elsawy, Gehan, Osama Abdelrahman, and Amr Hamza. "Effect of Choline Supplementation on Rapid Weight Loss and Biochemical Variables Among Female Taekwondo and Judo Athletes." Journal of Human Kinetics 40.1 (2014): 77-82.
• Herron, Kristin L., and Maria Luz Fernandez. "Are the current dietary guidelines regarding egg consumption appropriate?." The Journal of nutrition 134.1 (2004): 187-190.
• HSC: Homocysteine Studies Collaboration. "Homocysteine and risk of ischemic heart disease and stroke: a meta-analysis." Jama 288.16 (2002): 2015-2022. 
• Penry, Jason T., and Melinda M. Manore. "Choline: an important micronutrient for maximal endurance-exercise performance?." International journal of sport nutrition and exercise metabolism 18.2 (2008): 191.
• Ramstedt, Bodil, and J. Peter Slotte. "Membrane properties of sphingomyelins." FEBS letters 531.1 (2002): 33-37.
• Seshadri, Sudha, et al. "Plasma homocysteine as a risk factor for dementia and Alzheimer's disease." New England Journal of Medicine 346.7 (2002): 476-483. 
• Shin, William, et al. "Choline intake exceeding current dietary recommendations preserves markers of cellular methylation in a genetic subgroup of folate-compromised men." The Journal of nutrition 140.5 (2010): 975-980.
• Spencer, Melanie D., et al. "Association between composition of the human gastrointestinal microbiome and development of fatty liver with choline deficiency." Gastroenterology 140.3 (2011): 976-986.
• van Meurs, Joyce BJ, et al. "Homocysteine levels and the risk of osteoporotic fracture." New England Journal of Medicine 350.20 (2004): 2033-2041.
• Wu, Lily L., and James T. Wu. "Hyperhomocysteinemia is a risk factor for cancer and a new potential tumor marker." Clinica Chimica Acta 322.1 (2002): 21-28.
• Yates, Allison A., Sandra A. Schlicker, and Carol W. Suitor. "Dietary reference intakes: the new basis for recommendations for calcium and related nutrients, B vitamins, and choline." Journal of the American Dietetic Association 98.6 (1998): 699-706. 
• Zeisel, STEVEN H., et al. "Choline, an essential nutrient for humans." The FASEB journal 5.7 (1991): 2093-2098.

Betaine For Strength, Size & Endurance Gains: 2.5g/Day For 6 Weeks Does the Trick - Even in Advanced Trainees

If you feel like a shadow of yourself, betaine may help - specifically with  metabolically demanding workouts.

"Six-weeks of betaine supplementation improved body composition, arm size, bench press work capacity, attenuated the rise in urinary HCTL, and tended to improve power (p = .07) but not strength." (Cholewa. 2013) That's not just the conclusion to the latest paper Jason Cholewa et al. have just published in every supplement junkie's favorite scientific journal (do I have to mention it's the one of the International Society of Sports Supplementation, ISSN?), it's also exciting news on trimethylglycine and further evidence that it has the potential to queue up in the short line of effective dietary supplements you may spend money on without having a guilty conscience.

6 Weeks + 2.5g/day = Increased mass, volume and strength gains

The routine the researchers had their 23 experienced recreationally strength trained males (weight: 86.8 ± 9.1 kg; training experience: 4.8 ± 2.3 months; BF%: 16.9 ± 8%) between the ages of 18 and 35 follow was clustered into three microcycles (figure 1)

Figure 1: Overview of the training schedule sets, reps and rest times (Cholewa. 2013)

Obviously a pretty solid training program that was (no surprise) able to increase muscle size, strength and strength endurance irrespective of whether the subjects received

• When do you take betaine? After checking back with the author, I can now tell you that the betaine can be taken at any time point and that you may want to have it with a meal to avoid stressing your gastrointestinal system.
  2 x 1.25g placebo capsules, or
• 2 x 1.25g betaine capsules

The dosage was chosen because it's safe (9-12g/day = safe; ), effectively elevated plasma betaine levels (2.5-5g; ) and did already produce strength and performance gains in previous studies (Hoffman. 2009; Lee. 2010; Trepanowski. 2011).

Figure 2: Pre / Post bench press volume (left) and muscle cross section (right; Cholewa. 2013)

What is somewhat surprising, at least in my humble opinion, though, is the greater increase in bench press volume (figure 2, left) in the first microcycle and the "comeback" of the placebo group afterwards. Considering the fact that this parameter did not improve in previous studies at all, and comparing the workload on the chest day during the different microcycles, the researchers managed a reasonable explanation for this observation:

"Advanced Trainees Benefit from Increased Training Volume! Greater & Steadier Strength Gains with 8 Sets of Squats. Plus: Over 6 Weeks, 1 Set and 4 Sets Equally (In-)Effective." | read more

"Given improved work capacity with higher volume resistance training prescriptions, and the lack of improvement during micro-cycle 2 which imposed less of a metabolic demand (4 sets of 4–6 repetitions with 3 min rest), it is likely that betaine poses the most ergogenic potential in resistance training exercise protocolsthat impose higher metabolic demands.

Betaine is actively taken up by skeletal muscle during periods of stress, and may be ergogenic as an osmolyte by protecting sensitive metabolic pathways against cellular hypertonicity such as protein turnover, amino acid and ammonia metabolism, pH regulation, and gene expression. Specifically, betaine maintains cellular hydration to protect myosin ATPase and myosin heavy chain proteins against denaturation by urea. Moreover, the affinity of troponin for Ca 2+ , and thus force production, is negatively affected by reductions in protein hydration." (Cholewa. 2013)

Or put simply: The more metabolically challenging (~high volume) the workout the greater the chances you will benefit from taking betaine aka trimethylglycine (not be confused with betaine HCL, the digestive aid).

"Body recomposition" in trained athletes is what could betaine make a huge success

Against that background it seems only logical that the back squat work capacity improved nearly twofold compared to placebo during microcycle 3 (4 sets of 4–6 repetitions with 3 min rest; data not shown). You could even argue that the increases in muscle cross-sectional area (CSA) with betaine may occur only in the arms, because biceps and triceps are hammered directly and indirectly and thus at a relatively higher volume than the legs.

Figure 3: Changes in body composition in betaine & placebo group (Cholewa. 2013)

Eventually, the volume increase, or rather the improved handling of the metabolically demanding workouts, could also explain the highly desirable body recomposition effects we see in the betaine group. Less metabolic waste cluttering around = improved effects on body composition...? Well, the HCTL levels and changes in homocysteine the scientists measured would not support this notion. However,

Betaine content (in mg/100g) in some common food items (read more)

"[...] betaine supplementation may have impacted body composition via other mechanisms. Betaine has been shown to elevate plasma GH and IGF-1, and increase Akt phosphorylation in human skeletal muscle (Apicella. 2012). In mice betaine improves insulin sensitivity by restoring activation of IRS1 and the subsequent phosphorylation of PI3K/Akt by 50-100% in a concentration-dependent manner (Jakubowski. 2009). Thus, it is possible that by elevating anabolic hormones and enhancing downstream cellular signaling, betaine may have improved muscle protein synthesis, thus leading to an increase in lean mass." (Cholewa. 2013)

An alternative explanation Cholewa et al. present pertains to the osmo-regulatory effects of betaine, which may have lead to a "cellular swelling without an appreciable increase in myofibril protein accretion" (Cholewa. 2013). Obviously, these are not the gains you are looking for, but in view of the fact that Keller et al, were able to show that this correlates with decreases in proteolysis it would at least minimize the amount of protein that's getting lost from the musculature right after you "pumped" it into it (Keller. 2003).

---

What do eggs and homocysteine have in common? None of them causes heart disease. Eating eggs nay even prevent it by modulating the lipoprotein profile and cholesterol efflux (read more)

Bottom line: The study at hand adds more and, above all, highly relevant (human study, advanced trainees) evidence that betaine (2.5g/day in two servings) could make a valuable addition to the supplementation regimen of fitness models, physique competitors and bodybuilders, alike.

With its long-proven ability to reduce homocysteine levels () it has also been implicated as an agent that may prevent heart disease... unfortunately,  this assumption is based on the hypothesis that homocysteine plays a causative role in the development of heart disease and neither this nor the beneficial effects of choline & betaine on CVD are proven facts (Folsom. 1998; HCS. 2002; Olthof. 2005).

References:

• Apicella JM, Lee EC, Bailey BL, Saenz C, Anderson JM, Craig SA, Kraemer WJ, Volek JS, Maresh CM. Betaine supplementation enhances anabolic endocrine and Akt signaling in response to acute bouts of exercise. Eur J Appl Physiol. 2013 Mar;113(3):793-802.  
• Cholewa JM, Wyszczelska-Rokiel M, Glowacki R, Jakubowski H, Matthews T, Wood R, Craig SA, Paolone V. Effects of betaine on body composition, performance, and homocysteine thiolactone. J Int Soc Sports Nutr. 2013 Aug 22;10(1):39. [Epub ahead of print].
• Folsom AR, Nieto FJ, McGovern PG, Tsai MY, Malinow MR, Eckfeldt JH, Hess DL, Davis CE. Prospective study of coronary heart disease incidence in relation to fasting total homocysteine, related genetic polymorphisms, and B vitamins: the Atherosclerosis Risk in Communities (ARIC) study. Circulation. 1998 Jul 21;98(3):204-10.
• Hoffman JR, Ratamess NA, Kang J, Rashti SL, Faigenbaum AD: Effect of betaine supplementation on power performance and fatigue. J Int Soc Sports Nutr. 2009, 6:7.
• Homocysteine Studies Collaboration (HCS). Homocysteine Studies Collaboration. Homocysteine and risk of ischemic heart disease and stroke: a meta-analysis. JAMA. 2002 Oct 23-30;288(16):2015-22.
• Jakubowski H. The pathophysiological hypothesis of homocysteine thiolactone-mediated vascular disease. J Physiol Pharmacol. 2008 Dec;59 Suppl 9:155-67. Review.
• Lee EC, Maresh CM, Kraemer WJ, Yamamoto LM, Hatfield DL, Bailey BL, Armstrong LE, Volek JS, McDermott BP, Craig SA: Ergogenic effects of betaine supplementation on strength and power performance. J Int Soc Sports Nutr. 2010, 7:27.
• Keller U, Szinnai G, Bilz S, Berneis K. Effects of changes in hydration on protein, glucose and lipid metabolism in man: impact on health. Eur J Clin Nutr. 2003 Dec;57 Suppl 2:S69-74.
• Olthof MR, Verhoef P. Effects of betaine intake on plasma homocysteine concentrations and consequences for health. Curr Drug Metab. 2005 Feb;6(1):15-22.
• Trepanowski JF, Farney TM, McCarthy CG, Schilling BK, Craig SA, Bloomer RJ: The effects of chronic betaine supplementation on exercise performance, skeletal muscle oxygen saturation and associated biochemical parameters in resistance trained men. J Strength Cond Res. 2011, 25:3461–3471.

SuppVersity Science Round Up Seconds: Wheat Gluten Hydrolysates Fail, Exposure to Air Pollutants During Workout Reduces Brain Benefits, Homocysteine, B-Vitamins, Cognitive Impairment and Mortality

Before the profound weight loss (A) you don't see any of the glucose sucking and fad burning brown fat depots (black spots in B) on the neck of the in (B) 'foermerly obese', now only 'overweight' subject (also take a look at how the visceral fat in the abdominal region in (A) is actually pushing the organs upwards; img Vijgen. 2012)

Those of you who have listened to yesterday's show will have noticed that despite its flow the number of things you can discuss in a 1h podcast is simply very limited, to say the least. This is also why these Friday posts are probably never going to be simple summaries of the SuppVersity Science Round Up of the day before. The same is true for today and still I decided not to use the allegedly lame logo I did for the first two installments, but provide you with some 'real science' evidence of the absence of brown adipose tissue on the obese and it's magical reappearance after shedding 100lbs+ subsequent to a gastric bypass operation, instead (see image on the right).

Assuming that you have no idea what this "evidence" is for, I would suspect that you missed the live show yesterday and also did not find the time to download and listen to the podcast, yet -- right? Well, you should either download and listen to the show now and digest the Seconds later, or you read the following paragraphs first and download the podcast later.

What is not an option, however, is to miss one or another - I mean you can hardly want to eat the seconds if you have not had the main dish yet... and after listening to the podcast, I cannot imagine you don't want at least some seconds. Apropos seconds, here are today's seconds...

• Wheat gluten hydrolysate is not the new goto protein supplement - certainly not for female distance runners and probably not for anyone else, either! These are the kinds of studies that really annoy me. Studies that start out with blatant statements like "WGH [Wheat gluten hydrolysate] has been reported to suppress post-exercise rises in serum creatine kinase in male distance runners" (Hirao. 2012).
  
  

  

  Figure 1: CK, AST, ALT response in the "success trial" with men. In women even the miniscule beneficial effect on CK was not there. No reason to even think about buying a gluten hydrolysate as you new go-to protein supplement with only 5.6g of leucine/100g (whey has 50% more) and almost no GSH replenishing cysteine in it (0.9g vs. 3g+ in whey, which is more than +200% more).

  Sentences like that make the null-results of the study they precede look like the exception to the rule and are still nothing but a concession to a bias (let's hope not due to the grant from Nisshin Pharma Inc. which was the manufacturer of the wheat gluten hydrolysate used in this study). A bias, due to which an isolated observation as the slightly blunted increase in CK is blown up as if a slightly lower CK level was what could turn a sedentary pencil pusher into the next Hussein Bolt (Aoki. 2012).
  
  So, even if you are not afraid of the evil in gluten (which I believe not everyone has to), I strongly caution against making the switch from a high EAA protein with ton's of GSH boosting cysteine in it like whey to a mediocre grain protein, which is a potential allergen and contains tons of glutamine your body will readily turn into glucose, once it passes through the portal veign into the liver (I bet a large part won't even make it into systemic circulation).
  
  And as far as the purported "gender difference" goes the study at hand tries to blame the null result on (Hiriao. 2012), I suspect that it is rather the indisputable difference between the long-distance running at a continuous pace the women in the study at hand did, versus the totally different strains the guys in the previous study were exposed to during a soccer training + mini-match, which made the difference.
• Working out next to a street takes away some of the beneficial cognitive effects due to ultrafine particulate matter (UFPM) exposure. "Working out in the fresh air will promote weight loss more than working out inside." You heard me state that in one of the previous installments of the SuppVersity Science Round Up on Super Human Radio. Now this is still correct and based on sientific evidence, but at least as far as the cognitive benefits are concerned, working out outside does also have its downsides - at least for those of you who live in the inner city area.
  
  

  

  You better watch what you breath while you run.

  During a 12-week program the researchers from the Universiteit Brussel, the Hasselt University and the Royal Military Academy measured the improvements in physical performance, changes in serum markers and corresponding ultrafine particulate matter (UFPM) concentrations in the enviromnent in which their 15 previously untrained subjects conducted their aerobic training program thrice a week (Bos. 2012). What Bos et al. found was that the UFPM levels were signfificantly higher in the urban compared to the rural environment and that the higher UFPM exposures correlated with increases in leukocyte counts (p = 0.02), neutrophil counts (p = 0.04), and eNO levels (p = 0.002) that were exclusively observed in the group that trained in the urban environment.
  
  With the latter being markers of inflammation which exert their effects systemically, i.e. not just in the lung or musculature of which you may be thinking now, but also in the brain, it is no wonder that
  

  "reaction times on the Stroop task improved in the rural group (p = 0.001), but not in the urban group" (Bos. 2012). 

  What's comforting, though, is that the physical fitness did increase to a similar extend in both arms of the study.
• Homocysteine levels, mortality, cognitive impairment and which nutrients can offer some protection. I am not sure about what your impression is, but for whatever reason homocystein seems to be 'out of vogue' -- probably no room for it on the research agenda with all the hype surrounding vitamin D. It used to be all the rage in CVD risk research and today's news item is actually ain't about cardiovascular health, either.
  
  What the researchers from China and Taiwan actually were interested in was the correlation of high and low homocysteine levels with cognitive impairment and the corresponding nutrient intakes. In that Xiu et al. paid particular attention to the "B-vitamins" and found the following correlations between the mortality, cognitive status, homocystein levels and nutrient intake of their 1412 study participants (Xiu. 2012):
  

  • 

    Figure 2: Unadjusted mortality in the four quartiles of homocysteine levels (top); mortality according to homocysteine levels in subjects with different degrees of cognitive impairment (based on Xiu. 2012)

    if you go by the unadjusted data in figure 2, it's plain obvious that the all-cause mortality increases linearly from one quartile to the other 
  • this relation between plasma homocysteine levels and all mortality remained statistically significant after adjustments for age, sex, smoking status, BMI, physical function and general health were made
  • of the general foods, the scientists assessed, only regular fish intake had a statistically significant effect on homocysteine levels, with higher intakes being associated with lower homocysteine levels
  • of the b-vitamins choline was the only one with a significant association with plasma homocysteine levels (suggested read "Old School Supplement Choline Could Save Your Live and Liver!") 
  • neither betaine, nor vitamin B1, B2, B3 or B6 intakes did show statistically significant correlations with plasma homocysteine (not even "borderline significant; p > 0.15 for all, most way hither)
  • of the plasma markers, folate showed a highly significant correlation with homocysteine (14.4 nmol/L in the lowest HCY and 8.70 nnmol/L in the highest HCY group)
  • PLP, the active form of vitamin B6, came in close second with 70.3 nmol/l in the lowest HCY quantile and only 44.4 nmol / l in the highest quantile.
  Now, if you consider the fact that higher intakes of B-vitamins are probably not doing much to lower homocysteine levels int he elderly (at least not dose-dependently, when they are already getting enough) oddity #1, another look at the data in figure 2 will reveal oddity #2: The surprisingly high mortality in the lowest homocysteine quartiles in the patients with severe cognitive decline - how come? I mean, with low homocysteine they should not be at risk of having severe cognitive decline, anyway - right?
  
  Actually if you follow this rationale you can almost answer the question yourself. If you have low homocysteine and severe cognitive decline, the severe cognitive decline can hardly be from high homocysteine levels, so it must have another obviously pathological reason, or as the scientists have it
  

  "The joint effects of the 2 variables [homocysteine and cognitive decline] were most pronounced with severe cognitive impairment where mortality HRs ranged from 5- to 18-fold across a wide range of homocysteine concentrations. The findings with hypohomocysteinemia provide some insight into what might be an optimal range for this analyte in peripheral blood and tissues. The low concentrations may be seen with severe illness and malnutrition, and our study population comprises the health-vulnerable aged. For these reasons, we adjusted these associations for BMI (using the World Health Organization chronic energy deficiency category of, 18.5 kg/m 2 ), and we excluded those who died in the first year of follow-up. The findings were unchanged. Because mortality among the very old may have skewed the joint effects, these are presented for those ≤75 years and over, but again with similar findings." (

  A sarcastic person would now probably say: "We all have to go some time!" and just wave his hands at these results. True! And I am the last to advice you to become over-anxious. Yet in the mean time it would appear prudent to make sure to get your homocysteine levels checked from time to time, not to forget that choline is a b-vitamin as well and not to fall for the idea that you cannot overdose on B-vitamins - I don't have to remind you of the negative effects, specifically folic acid supplementation can have on all sorts of cancer (e.g. breast cancer, where a high folic acid intake from foods and supplements is associated with a +30% risk of cancerous growth; cf. Kim. 2006).

In case you are looking for the post on "ammonia accumulation brain-fog, toxicity, liver 'pathologies' and workout performance", yeah it was on the list, but I decided it would be a shame to tackle that within a short two paragraph seconds items. Don't worry I am not going to forget about it, after all its in my humble opinion one of the main reasons the diets and workout regimen of the many ambitious physical culturists fail. If you are still looking for more and have not listened to the podcast, yet, this would be the right moment to download the file from the Super Human Radio Network server (click here to download), otherwise the latest short news on the SuppVersity Facebook Wall may offer some diversion ;-)

References:

• Aoki K, Kohmura Y, Suzuki Y, Koikawa N, Yoshimura M, Aoba Y, Fukushi N, Sakuraba K, Nagaoka I, Sawaki K. Post-training consumption of wheat gluten hydrolysate suppresses the delayed onset of muscle injury in soccer players. Exp Ther Med. 2012 Jun;3(6):969-972. Epub 2012 Apr 3.
• Bos I, De Boever P, Vanparijs J, Pattyn N, Panis LI, Meeusen R. Subclinical Effects of Aerobic Training in Urban Environment. Med Sci Sports Exerc. 2012 Oct 15.
• Cankurtaran M, Yesil Y, Kuyumcu ME, Oztürk ZA, Yavuz BB, Halil M, Ulger Z, Cankurtaran ES, Arıoğul S. Altered Levels of Homocysteine and Serum Natural Antioxidants Links Oxidative Damage to Alzheimer's Disease. J Alzheimers Dis. 2012 Oct 29.
• Guest PC, Urday S, Ma D, Stelzhammer V, Harris LW, Amess B, Pietsch S, Oheim C, Ozanne SE, Bahn S. Proteomic analysis of the maternal protein restriction rat model for schizophrenia: Identification of translational changes in hormonal signalling pathways and glutamate neurotransmission. Proteomics. 2012 Oct 16.
• Hirao T, Koikawa N, Aoki K, Sakuraba K, Shimmura Y, Suzuki Y, Sawaki K. Female distance runners show a different response to post-workout consumption of wheat gluten hydrolysate compared to their male counterparts. Exp Ther Med. 2012 Apr;3(4):641-644.
• Kim YI. Does a high folate intake increase the risk of breast cancer? Nutr Rev. 2006 Oct;64(10 Pt 1):468-75.
• Vijgen GH, Bouvy ND, Teule GJ, Brans B, Hoeks J, Schrauwen P, van Marken Lichtenbelt WD. Increase in brown adipose tissue activity after weight loss in morbidly obese subjects. J Clin Endocrinol Metab. 2012 Jul;97(7):E1229-33. Epub 2012 Apr 24.
• Xiu LL, Lee MS, Wahlqvist ML, Chia-Yu Chen R, Huang YC, Chen KJ, Li D. Low and high homocysteine are associated with mortality independent of B group vitamins but interactive with cognitive status in a free-living elderly cohort. Nutr Res. 2012. Ahead of print.

Old School Supplements - Choline: Stronger, Faster, Leaner & More Muscular, or Just Another Dumb-and-Barbell Story?

Image 1: You really get a hefty dose of the history of physical culture with volume 1 + 2 of Randy Roach's Muscle, Smoke & Mirrors; I must forewarn you, though: These books have addictive potential, so don't buy them, when you have other important things (beside training ;-) on your schedule.

If you are among my facebook friends, you will probably remember that I have received an amazing gift from my friend Carl Lanore, a few weeks ago: The first two books of Randy Roach's Muscle, Smoke & Mirrors (soon-to-be) Trilogy (Roach. 2008-2011). I am not a fan of muscle gossip, but these books are a really amazing resource for everyone with a vested interest in physical culture - just as Super Human Radio the "first radio station dedicated to promote physical culture is, by the way ;-)

My personal favorite in the first two volumes of Muscle Smoke & Mirrors is yet clearly the last third of the second volume about the history of dietary supplements. Aside from the notorious liver tabs and all sorts of "pro-diarrheal" protein supplements, stories about Joe Weider being willing to sell elephant shit, if only someone convinced him that it would further protein anabolism, and a lot of other informative and entertaining stuff, there was one supplement that has been largely forgotten by now that caught my interest - choline!

Choline is present in every human cell!

Although it does not carry the term "vitamin" in its name, choline is an essential water-soluble nutrient that is abundant in some of the typical old-school bodybuilding foods, such as calf liver, beef, eggs, chicken, turkey, sardines, cod, milk and a lot of those green vegetables, many people today believe they must have been invented by the vegans and vegetarians.

Are vitamin supplements bad for me? I have addressed the question in some detail in two previous blogposts in response to the ever resurfacing horror stories about vitamin E, selenium and prostate cancer (Part I), and the media hype around the results of the Iowa Women's Health Study (Part II). So before you bombard me with further questions, go and check out these and a couple of posts on hormesis, here at the SuppVersity ;-)

At first sight it may thus seem that the supplemental choline some of the pros took with each and every meal was at best a waste of time and money, if not a potential health hazard; after all, we are seeing all those horror news about potential negative side-effects from the overconsumption of all sorts of allegedly harmless vitamins pop up in the media in ever shorter time intervals, as of late.

Choline - essential, but already (over-)abundant?

To answer this question we will initially have to identify what choline does in our bodies. As the name that has been derived from the greek word for bile ("chole") implies, choline has been initially identified as a major component of the juices and cells of the pancreas and liver. Scientists realized only later that it is an absolutely essential constituent of each and every cell of our body, where choline with its fat-modifying properties increases the flexibility of the cell membranes and handles the in- and outflux of fat-based nutrients and waste products, respectively.

If we take into consideration that choline is also one of the rare trimethylated molecules in our diets and acts as an important methyl donor that is required for both, the proper activation and deactivation of genes, and as precursor to the neurotransmitter acetylcholine, it is hardly surprising that the list of mostly neurological and cardiological pathologies stemming from marginal or full-blown choline deficiency is endless:

Image 2: The purportedly dangerous cholesterol bombs, aka eggs, are among the #1 sources of dietary choline

• high homocysteine levels, cardiovascular disease, 
• high blood pressure, high triglyceride levels
• fatigue, insomnia, 
• memory and nerve problems,
• liver dysfunction, kidney failure
  (probably subsequent to a lack of phosphatidylcholine),
• impaired growth and failure to thrive,
• abnormalities in bone and red blood cell formation,
• infertility

If you take a look at your "scientifically formulated high potency B-vitamin supplement" *rofl* you may notice that despite getting 1000%-3000% of all sorts of B-Vitamins, chances are that it contains "only" 100% of the recommended daily allowance of choline (USDA recommendations):

Figure 1: The daily choline intake of most Americans is below the respective RDA, of which some scientists already speculate that it may already be too low. (USDA. 2011)

• 0-6 months: 125 mg
• 6-12 months: 150 mg
• 1-3 years: 200 mg
• 4-8 years: 250 mg
• males 9-13 years: 375 mg
• males 14 years and older: 550 mg
• females 9-13 years: 375 mg
• females 14-18 years: 400 mg
• females 19 years and older: 425 mg
• Pregnant females of any age: 450 mg
• Lactating females of any age: 550 mg

And though the "average" American today is working his way up towards a similar "weight class" as the heavy weight bodybuilders in Arnold's days, the disproportionally higher muscle mass of a bodybuilder as well as the 8,000-10,000kcal diets those big guys were consuming in the off season would suggest that they needed at least twice, maybe even thrice the amount of choline an average individual would consume. The "OSBDA", as in old-school bodybuilding daily allowance would thus have been roughly 1.5g of dietary choline, as it is contained in

• 15 eggs
• 10-11 eggs and 1lbs of chicken
• 1 cup of almonds, three cups of rice, 5 eggs, 1lbs of chicken and five ounces of liver

And while many of the pros probably got way more than those 1.5g of choline from their glutenous and ridiculously frequent meals, the increased oxidative damage due to the arduous workouts and use of certain "supplements" may well have exacerbated their need for a nutrient of which various epidemiological studies report a significant correlation with reduced levels of C-reactive protein (CRP), interleukin-6 (IL-6) and the tumor necrosis factor-alpha (TNF-alpha); the usual suspects, all of which  have been implicated as confounding, if not causative factors in almost every modern (e.g. Detoupolo. 2008, Rajaie. 2011).

Bigger, stronger, faster, leaner, ... what the pros said

For most, yet by probably not all competitive bodybuilders back in the day, improvements in heart and overall health were yet probably not the major incentive to ramp up their choline intake to levels, where the fishy smell your sweat, urine and other bodily fluids will develop when you ingest way too much choline eventually drowned out the foul protein farts, they got from the hardly digestible protein powders of the late 1970s. What they were looking for was the competitive edge:

1. Performing more reps and training at an even higher volume - With dietary / supplemental choline being a necessary precursor to acetylcholine, which in turn facilitates skeletal muscle contraction, it was only reasonable to assume that the 2-3x per day 2-3h marathon workouts some of the guys were performing, would increase the risk to deplete your choline levels so that the acetylcholine production would come to a standstill and your muscles would simply refuse to contract.
    
2. Greater protection and faster repair of muscle damage - Due to its established anti-oxidant effects and its status as an essential and functional constituent of the cell wall, it appears logical that adequate levels of choline would be necessary to both protect and repair skeletal muscle.
    
3. Increased leanness and vascularity - In view of the fact that it has been known since the early 1950s that choline's role in the oxidation of fatty acid goes well beyond serving as a source for the phospholipids that would carry them out of the liver and to the mitochondria of skeletal muscle and other metabolically active tissue (Artom. 1953). Even the assumption that choline supplementation will propel the oxidation of fatty acids, promote lower body fat levels and lead either directly (fat loss) or indirectly (reduced inflammation = reduced subcutaneous water) to a more vascular look, was far from being a bro-scientific 'dumb-and-barbell story'.

As you as an educated SuppVersity student know all too well, though, not everything that appears to make sense works and not everything that that works must necessarily have scientific research to back its efficiacy up - after all, the brocebo effect alone (cf. "Add 10kg to Your Bench With Brocebos") could have been responsible for a subjective decrease in fatigue (1), a decrease in muscle soreness (2) and a perceived increase in leanness and vascularity (3).

... what exercise and nutrition science says

Despite the last-named restraints and the unlikeliness that we will find a study on choline supplementation on 200lbs+ bodybuilders on a 8,000kcal/day+ diet, I guess, you will not mind, if we take a brief look at whether or not those "logical" benefits (#1-3, above) have actually been observed in peer-reviewed studies.

1. Potential effects of supplemental choline on training load / volume Van Allwörden et al. weer among the first scientists who tried to establish a connection between the long-established exercise induced decreases choline, exercise performance and fatigue (Van Allwörden. 1993). Just like Buchman et al. who report similar results in marathon runners (Buchman. 2000), Allwörden et al. had yet to acknowledge that the 0.2g/kg lecithin (a choline source) induced compensation of the 17% decrease in serum choline levels compared to the control group did not lead to increase the performance of the adolescent triathletes in their study. And though these results stand in contrast to a 1992 study by Sandage et al. which found a minimal increase in 10k running times in response to the ingestion of 2.8g of choline 1h prior to the run, most authors of pertinent reviews do acknowledge the theoretical merit of the hypothesis, but speculate that the decline in choline is rarely ever pronounced enough to for choline supplements to illicit immediate ergogenic benefits (e.g. Jäger. 2007; Penry. 2008). Long-term studies on potential downstream effects of decreased oxidative stress, improved cellular regeneration and utilization of fat soluble nutrients, on the other hand, are missing so that the preliminary answer to the question whether choline supplementation would allow you to train longer, or at an overall higher volume must be: Very unlikely!
    
2. Potential effects of supplemental choline on muscle damage, repair and growth While Michel et al. observed in isolated muscle cells that choline deficiency induced a cascade of changes in both the oxidative metabolism (downregulated), as well as the fatty acid composition of the cell membranes (Michel. 2011), which was characterized by a shift towards mono- and away from saturated fatty acids, it is somewhat far-fetched to use this as "evidence" for a potential beneficial effect of choline supplementation; after all, we are dealing with isolated, choline depleted muscle cells in a petri dish and cannot even say for sure whether a potentially hightened susceptibility of the monounsaturated fats in the cell wall will lead to an increase in skeletal muscle damage in response to strenuous exercise. Against the background that there is no other convincing scientific evidence that would suggest that supplemental choline - in the absence of dietary choline deficiency (or pathologies related to the latter, such as liver cirrhosis) - would make your muscle bullet proof, this is the 2nd purported benefit the real-world significance of which turns out to be more than questionable.
    
3. Potential effects of supplemental choline on fat loss and vascularity Interestingly enough, the "fat loss" hypothesis, appears to be the one with the most convincing scientific evidence to support it. In 2002, for example, Hongu and Sachan observed a shift towards increased fatty acid oxidation in 19 healthy women who participated in a combined carnitine + choline + exercise trial. Moreover, these substrate repartitioning effects were "sustained until wk 2 after cessation of choline plus carnitine supplementation and exercise" (Hongu. 2002). In a previous rodent trial a similar stack that included caffeine, carnitine and choline had lead to body fat reductions "similar to those due to mild exercise" (Hongu. 2000), an observation that would support a previous hypothesis of the authors stating that the combination of carnitine and choline favors an "incomplete oxidation of fatty acids and disposal of their carbons in urine as acylcarnitines in humans" and could thusly help to increase the absolute and relative amount fat loss on a diet. In view of the well-established lipolytic effects of caffeine a "CCC stack", i.e. caffeine + carnitine + choline, could thus in fact make a valuable addition to a sound exercise and diet regimen. Whether it would really make a visible difference, remains yet questionable.

What is the take away message from this article then? I guess, the best answer to this question would be eat your eggs, (organ-)meats, fish, nuts and veggies (and to avoid soy lecithin in supplemental or any other form) to satisfy your choline requirements naturally, in order to remain the "metabolically healthy, fat burning, muscle building machine" you should by now be with all those daily tips on health, exercise, nutrition and supplementation, here at the SuppVersity ;-)

References:

• von Allwörden HN, Horn S, Kahl J, Feldheim W. The influence of lecithin on plasma choline concentrations in triathletes and adolescent runners during exercise. Eur J Appl Physiol Occup Physiol. 1993;67(1):87-91.
• Artom, Camillo. Role of Choline in the Oxidation of Fatty Acids by the Liver. J. Biol. Chem. 1953 205: 101-111. 
• Buchman AL, Awal M, Jenden D, Roch M, Kang SH. The effect of lecithin supplementation on plasma choline concentrations during a marathon. J Am Coll Nutr. 2000 Nov-Dec;19(6):768-70.
• Detopoulou P, Panagiotakos DB, Antonopoulou S, Pitsavos C, Stefanadis C. Dietary choline and betaine intakes in relation to concentrations of inflammatory markers in healthy adults: the ATTICA study. Am J Clin Nutr. 2008 Feb;87(2):424-30. 
• Hongu N, Sachan DS. Caffeine, carnitine and choline supplementation of rats decreases body fat and serum leptin concentration as does exercise. J Nutr. 2000 Feb;130(2):152-7.
• Hongu N, Sachan DS. Carnitine and choline supplementation with exercise alter carnitine profiles, biochemical markers of fat metabolism and serum leptin concentration in healthy women. J Nutr. 2003 Jan;133(1):84-9.
• Jäger R, Purpura M, Kingsley M. Phospholipids and sports performance. J Int Soc Sports Nutr. 2007 Jul 25;4:5. 
• Penry JT, Manore MM. International Journal of Sport Nutrition and Exercise Metabolism 2008, 18(2):191-203
• Rajaie S, Esmaillzadeh A. Dietary choline and betaine intakes and risk of cardiovascular diseases: review of epidemiological evidence. ARYA Atheroscler. 2011 Summer;7(2):78-86.
• Roach, Randy. Muscle Smoke & Mirrors. Volume 1-2. AuthorHouse. 2008-2011.
• Sachan DS, Hongu N. Increases in VO2max and metabolic markers of fat oxidation by caffeine, carnitine, and choline supplementation in rats. J Nutr Biochem. 2000 Oct;11(10):521-6. 
• Sandage BW, Sabounjian RN, White R, Wurtman RJ: Choline citrate may enhance athletic performance. Physiologist 1992, 35(4):236. 
• USDA. FSRG Dietary Data Brief -- No. 9 –Dietary Intakes of Choline. 2011

Intermittent Thoughts on Intermittent Fasting - The Fast #2: Health & Longevity Effects of Intermittent Fasting

Image 1: Any roundworms reading this? I hope you know that by not eating enough and feeling miserable you can extend your lifespan ;-)

Thanks Caenorhabditis elegans, or "C. elegans", almost everybody who is able to read a newspaper or online magazine will have heard of the miraculous effects of calorie restriction and fasting on longevity (of this worm!). Since I assume that you possess more gray matter than this transparent nematote, you will probably have asked yourself how, or rather if these results from a worm with an average lifespan of 2-3 weeks translate to human beings,... well, all I can tell you is that leading a miserable life of lifelong dieting appears to work in non-human primates, as well (Kemnitz. 2011). Now, the obvious question is:

How does all that relate to Intermitent Thoughts on Intermittent Fasting? Obviously, none of the roundworms or rhesus monkey's followed Martin Berkhan's intermittent fasting approach, did they?

No, I have not seen pictures of abes on leangains.com, either, so I suppose they did not follow Berkhan's approach, but - and this is the likewise fascinating, as well as surprising, connection - there is scientific evidence that intermittent fasting could reproduce some of the beneficial effects of caloric reduction while avoiding a whole host of its undesirable side-effects. Reason enough for me, to devote this episode of the increasingly popular Intermittent Thoughts on Intermittent Fasting series to the non-cosmetic health effects of intermittent fasting (I know that's not as getting ripped and jacked, guys ;-)

Figure 1: Ramadan fasting leads to profound beneficial changes in inflammatory markers (left) and blood lipids (right) in 20 healthy male non-obese volunteers aged 23-39 (data calculated based on Aksungar. 2006)

We touched on one of this unsexy, yet vitally important health-benefits at the end of the last installment on the metabolic and endocrine effects of fasting, already: the beneficial effect on insulin resistance (as measured by a +44% increase in the inverse of the long-term marker of insulin resistance 1/HOMA-IR and the slight increase in QUICKI) Shariatpanahi et al. had observed in 55 ramadan fasting subjects (Shariatpanahi. 2008) for example constitutes one of these "boring side-effects", of which many dieters fail to realize that improvements in insulin sensitivity or reductions in inflammatory markers and C-reactive protein (cf. figure 1, left), as they were reported by Aksungar et al. in a 2006 study on the effects of religious "intermittent" fasting (Aksungar. 2006) facilitate weight - and more specifically - fat loss.

Image 2: Would the issue of dehydration distort the results of Ramadan studies and thus render them irrelevant for our argumentation? (photo Offline Clinic)

As in almost every episode I want to make a few brief statements regarding the "Ramadan model of intermittent fasting". I have already discussed its advantages over "over-other day" or "alternative day" fasting in the second installment of this series and I have hinted at possible problems related to the restriction of water intake within the fasting period in the third installment of the series. Now, in view of the general recommendation to drink more and more frequently to avoid dehydration (USDA), it seems prudent to verify that potential beneficial health effects of fasting are not masked by detrimental side effects of dehydration, if we - once again - want to rely on the available data on religious fasting as part of our argumentation. In a paper published in the European Journal of Clinical Nutrition in 2003, Leiper, Molla and Molla report that despite the fact that "[d]uring the daylight hours of Ramadan fasting, practising Muslims are undoubtedly dehydrating", it is neither "clear whether they are chronically hypohydrated" nor have there been any "detrimental effects on health [...] directly attribut[able] to negative water balance at the levels that may be produced during Ramadan" observed in any scientifically relevant studies (Leiper. 2003). It is thus relatively safe for us to assume that we can neglect he influence of potential dehydration in our initial analysis of respective studies.

Now, what about the longevity effects?

Image 3: The effects the calorie restriction had on the ape on the right is certainly impressive, when you compare it to his 27.6 year old age-mate on the left - would be interesting to see how an "intermittently fasted version would end up ;-) (img. from the Irish Medical Times)

With or without the beneficial effects on cardiovascular risk markers, the question still remains: "Can intermittent fasting mimic the longevity effect of calorie restriction without the constant cravings, continuous hunger and all sorts of "human" problems that won't show in studies on yeast, worms, rodents or even apes (see image 3)?"

In order to answer this question, we will first have to give a clear cut definition of what our understanding of intermittent fasting is, in this context, because, obviously, if we coupled our (intermittent) fast with a a deliberate and severe calorie restriction we would probably end up like Canto, the ape from the the longevity study in image 3 - old, but miserable. What we want, on the other hand, is an intermittent fast, where - within the feeding window - we are allowed to eat to satiety... similar to what we are seeing in religious fasting: no calorie counting and perceived (when compared to normal meal sizes) overeating.

Now, we do already know from the studies cited above (and in previous installments of this series) that one month on a dietary regimen like that provides considerable benefits as far as weight loss, insulin sensitivity, blood lipid ratios (! this is important, because we are knowledgeable enough to give a f*** about total cholesterol & co) and inflammatory markers, all of which are associated not only with an increased life-expectancy (Danaei. 2010), but, more importantly, with an increased life-quality and neurological health (Bronwen. 2006) up into the old(er) ages. From the same studies, we do yet also know that breaking the fast, and returning to our old dietary habits returns these markers to baseline (cf. post values in figure 1). What we do not know, however, is whether ...

1. intermediate improvements in correlates of health and longevity as they can be observed during intermittent periods of Ramadan (intermittent) fasting provide any, or even significant longterm health benefits, and
2. longterm (as in for years and decades) intermittent fasting would not over months or years lose its effect, or even worse, be detrimental to our overall health and thus reduce not extend our life-expectancy and/or life-quality

From rodent studies we already know that a life-long alternate-day feeding protocol increases lifespan in the absence of any caloric reduction (Goodrick. 1990; Mattson. 2000). Evidence that similar beneficial effects, at least as far as mortality from chronic disease is concerned, would occur in human beings comes from a 2007 study by Varaday and Hellerstein (Varaday. 2007), who do yet remark that (as I already pointed out) "more research is required to establish definitively the consequences of ADF [alternate day fasting]".

The circadian clock hypothesis

A finding from alternative day fasting studies that may be of paramount importance in view of the "quality of life"-aspect, is the increase in brain-derived neurotropic factor (BDNF) that has been observed in animal studies. BDNF is involved in brain development and plasticity and its (intermittent) fasting-induced elevation could explain the neuroprotective effect of respective feeding patterns (Duran. 2001). In a review of the literature, Fory and Miskin do yet remark that (Froy. 2010)

[...] BDNF could not be [directly] linked to the neuro-protective effects in the brain of calorically restricted rats, but increased levels of another neurotrophic factor, glial cell line-derived factor (GDNF), were correlated with neuro-protection of a calorically restricted primate model of Parkinson's disease. Interestingly, BDNF is also a component of the hypothalamic melanocortin pathway that controls food intake and body weight in adult mice, and it has been implicated in the regulation of energy metabolism.

After all, the scientists believe that IF exhibits part or even all of its effects by (re-)setting clock genes, a hypothesis which despite having its merits would yet lead us into theoretic considerations with little merit to our "intermittent thoughts" on the real-world outcomes of intermittent fasting, which is why I will, at this point, skip forward to the results of a 1999 study into the relation of adipose tissue size and reductions thereof to longevity.

Is it not about eating less, but just about getting leaner?

Image 4: When we are talking about the benefits of losing body fat, this obviously does not imply you have to get in Phil Heath Mr. Olympia '11 shape to live longer (photo bodybuilding.com)

In the introductory paragraphs of this installment of the Intermittent Thoughts on Intermittent Fasting series I somewhat ridiculed the idea of changing one's whole life to get "ripped and buffed", now, after revisiting some of the studies and observing the close correlation of improvements in metabolic health markers, reductions in body fat and consequent life-extending effects of (intermittent) fasting, the central question of Nir Barzilai's and Gaurav Gupta's 1999 paper Revisiting the Role Fat Mass in the Life Extension Induced by Calorie Restriction seems by no means far fetched: What if ain't inflammation and insulin resistance & co that make us fat and reduce our lives, but being fat that leaves us insulin resistant, inflammed & co and thusly reduces our life-expectancy? The answer, according to Barziilai and Gaurav, is simple (Barzilai. 1999):

In fact, all of the benefits of CR on the neuroendocrine system and those related to the improvement in glucose homeostasis can be attributed to decrease in adipose cells and their products.

Probably too simple and above all difficult to treat with a drug and thus not profitable enough to be accepted by the medical establishment. Now, if this would be the case, intermittent fasting would in fact provide the sought-after silver bullet to leading a leaner, healthier life, as both the anecdotal reports on the Net, as well as the the majority of the studies that have been cited in this, as well as the previous installments show quite conclusively that going without food for periods <24h is capable of reducing body fat stores, while preserving lean muscle (and bone) mass. But why, or better how does that work?

Intermittent fasting, fat reduction, health improvements and the cyclicality of life

Illustration 1: It probably is the magic interplay of AMPK and mTOR that produces such remarkable transformations as Duong Nguyen's. We will delve deeper into the their reciprocal interaction in the next installment of the Intermittent Thoughts on Intermittent Fasting Series

Due to its cyclic nature intermittent fasting seems to temporarily produce similar "reductions in protein synthesis" (Barrows. 1978) as the ones which have been established as the fundamental mechanisms of the life-extending effects of calorie restriction as early as in the late 1970s and of which we know today that they partly mediated by an AMPK-induced downregulation of the mTOR pathway. In that, this transient AMPK response to intermittent fasting, which most intermittent fasters further stimulate by exercising in a (semi-)fasted state, appears to be profound enough to stimulate or, according to some recent research (cf. Canto. 2010), we should probably say "sustain" the SIRT1-pathway to an extent that allows dieters to benefit from its immediate fat-mobilizing effect on white adipocytes (Picard. 2004) and its ability to maintain telomer length (and subsequent longevity, cf. Palacios. 2010) without the unwanted muscle-wasting side-effects of prolonged fasting periods.

The fasting induced AMPK expression is yet only one part of the cycle, of which we have learned in this installment that it is responsible for both, the "cosmetic" effects on body fat, which could, after all, be causative and not just corollary or even subsequent to improvements in insulin sensitivity (cf. What Comes First: Inflammation or Obesity?), lipid profiles, and the whole string of beneficial health effects which irrefutably contribute to the longevity effect of inhumane low-calorie diets. In view of the fact that another valuable Sunday afternoon is drawing to a close, the discussion of the second player in this cycle, the mammalian Target of Rapamycin, or in short mTOR, will yet have to be postponed to the next episode of the Intermittent Thoughts on Intermittent Fasting Series - so stay tuned, keep the questions and comments coming and don't forget to check back on Thursday to see what AMPK and mTOR are doing to our man at the 2012 wheelchair nationals, Adelfo Cerame!