Science Round-Up Seconds: 8 Nootropics to Combat Stroke, Alheimer's & Co and Boost Cognitive Performance. Plus: 7 Rarely Thought of Side Effects of High Dose Glutamine.

Effects of infusion times on phenol content of black tea (Ramalho. 2012)

If you have already listened to the podcast of yesterday's Science Round-Up on the Super Human Radio Website (click here if you haven't and wan't to know what the following is all about), I suppose you will not mind that I compiled some of the complex information about "optimal" tea brewing in the illustration to the right (based on Ramalho. 2012). The colored arrows indicate the time-points at which the given compounds in the tea achieved peak values. The exact time point is also given in minutes, so that a 9' in front of the green caffeine and on the left to the green arrow pointing at the 9 min point tells you "it took 9 minutes for the caffeine content to reach it's maximum in the British tea". The graph in the background shows the catechin concentration depending on the infusion time.

Cholinergenic nootropics - What a recent review says

I guess some of you will probably have heard about piracetam or lecithine as purported enhancers of cognitive function. According to a recent review in the Journal of Experimental Pharmacology those two are yet not the most prosing agents:

Eggs are rich in choline which is an essential nutrient and was abundant in the classic BB diets (rear more)

• Piracetam: no cerebroprotective effects in patients who have open heart surgery, but does help on non-open cardiopulmonary bypass surgery (Holinski. 2008), beneficial effects in response to cerbrovascular and cognitive disorders traumatic origin (Malykh. 2010), intravenous piracetam can prevent cognitive deficits in response to anesthesia (Fesenko. 2009)

• Lecitin: does not improve cognitive deficits in patients (Amenta. 2011; Parnetti. 2007)

More promising "nootropics" - specifically in view of what most people do actually expect, when they buy such products.

• Oxiracetam: improves cognitive performance except for patients with dementia (Malykh. 2010)

• Citocoline: general neuroprotective effects (Alvares-Sabin. 2011), improvements in cognitive performance in healthy and patients and patients with dementia (Secades. 2010), helps with cognitive dysfunction in Parkinson's (Vale. 2008), helps with cognitive function in dementia of neurodegenerative and vascular origin (Parnetti. 2007), prevents cognitive decline after a stroke (Alvarez. 2011), improves recovery after stroke (Garcia-Cobos. 2010) 

• Cerebrolysine: produces signifant cognitive improvements in vascular dementia (Guekht. 2011), effective for both cognitive function and behavioral symptoms in Alzheimer's (Alvarez. 2011), promising results in patients with Alzheimer's (Plosker. 2009)

And a couple of things you would not usually associate with nootropics:

• 

  Suggested read: Amino Acids for Super Humans on the effects and differences between the various forms of carnitine (read more).

  Acetyl-L-carnitine: improves cognitive performance in patients with encephalopathy, decreases anxiety and increases general energy and wellness, as well as fatigue and age-related cognitive deficits (Malaguernera. 2008, Liu. 2008),can reduce or block neuronal death in neurodegenerative diseases (Manusco. 2007), helps ammeliorate hyperammonemia (Cagnon. 2007)

• Saffron extract: beneficial effects in mild to modest Alzheimer's  (Akhondzadeh. 2010)

• DHA (fish oil): positive effects on verbal recognition memory in old subjects (Yurko-Mauro. 2010)

Interestingly, the most profound effects appear to be brought about by acetyl-l-carnitine. In that it's worth mentioning that the benefits could still be related to cholinergic mechanisms, since it has long been known that ALCAR can increase the expression of choline acetyltransferase activity in the central nervous system (Taglialatela. 1994). And the latter is, as the name implies, necessary to form the neurotransmitter acetylcholine .

Glutamine probably not suitable for chronic high dose supplementation

Czech scientists warn about the risks of chronic high dose glutamine supplementation. I know that many of you are still too bamboozeled by the "protein for everything and let the liver take care of any glucose demands I may have" theory, of which you could probably argue that it is the bastard child of the standard BB diet with low carb. Maybe the following recently published paper by a scientist from the Charles University in Prague can help cure this "disease" (and your cognitive problems, fatigue and brainfog).

According to Holecek, the chronic ingestion of glutamine / glutamine enriched diets in can lead to...

Figure 1: In the presence of high amounts of glutamine outside of the cell, the glutamine synthesis (GLN) and with it the ammonia detoxification from muscle tissue sucks (Holecek. 2012).

"(1) Alterations in amino acid transport-as GLN shares the transporters with other amino acids, enhanced GLN intake may impair amino acid distribution among tissues and their absorption in the gut and kidneys.

(2) Alterations in GLN metabolism-GLN supplementation may impair synthesis of endogenous GLN and enhance glutamate and ammonia production.

(3) Alterations in ammonia transport-GLN supplementation may impair ammonia detoxification and negatively affect the role of GLN as the carrier of ammonia among tissues.

(4) Abnormalities in aminoacidemia-increased plasma levels of GLN, glutamate, citrulline, ornithine, arginine, and histidine and decreased levels of valine, leucine, isoleucine, glycine, threonine, serine, and proline are reported.

(5) Alterations in immune system-as GLN has immunomodulating properties, the effect of chronic GLN consumption on the immune system needs to be assessed.

(6) Effect on tumor growth-it should be elucidated whether chronic intake of GLN increases the risk of cancer.

(7) Effect of the withdrawal of GLN supplementation-due to the adaptive response of the organism to enhanced GLN consumption, the withdrawal of GLN may enhance the risk of health problems resulting from GLN deficiency." (Holecek. 2012)

Remember the post on the ammonia induced peripheral and central fatigue with high dose chronic BCAAs supplementation?

In view of the fact that some people consumer up to 40g of glutamine regularly, Holecek demands that "long-term studies should be performed" to test the side effects and evaluate whether there is any benefit at all to justify chronic consumption of a GLN-enriched diet.

So, relying on glutamine instead of carbs, as smart as this idea appears to be in the current carbophobia, could actually make you stupid due to the disruption of the intracellular ammonia detoxification, which is not a problem in muscle only, but also in the brain.

In the end, what we are seeing here is just another instance of a disruption in the natural balance of things. Ornithine, citrulline and arginine, for example are involved in the detoxification of ammonia via the urea cycle. They are however not the only bottleneck to the system.

Obviously your liver and kidneys will have to handle the clearance. People with liver problems (or persons taking "supplements" or NSAIDs that may impair the liver function) are therefore particularly prone to hyperammonemic encephalopathy (Kanamori. 1996; Lemberg. 2009)

Bottom line: Glutamine, just like everything else, in moderation and by no means so much that your body runs on glutamine as fuel. Aside from the mentioned amino acids that help the clearance of ammonia from the blood stream, taurine appears to exert a direct protective affect in the brain (Chepkova. 2006), and lactulose (a fermentable carbohydrate) can reduce the ammonia influx from ammonia producing bacteria in the gut (Vince. 1980). So if you want to wear a helmet when you bang your head against the wall, these would be suggested "take supplement B in order to counter the side effects of supplement A" - side effects of a supplement you would not even have to take, by the way (100% bro-logic ;-)

References: 

• Amenta F, Carotenuto A, Fasanaro G, Lanari A, Rea R, Traini E. Preliminary results of Ascomalva trial on the association of donepezil and choline alphoscerate in Alzheimer’s disease with associated cere-brovascular injury. G Gerontol. 2011;59:89–9.
• Akhondzadeh S, Shaf iee Sabet M, Harirchian MH, Togha M. A 22-week, multicenter, randomized, double-blind controlled trial of Crocus sativusin the treatment of mild-to-moderate Alzheimer’s disease. Psychopharmacology (Berl). 2010;207:637–643.
• Alvarez XA, Cacabelos R, Sampedro C, et al. Efficacy and safety of cerebrolysin in moderate to moderately severe Alzheimer’s disease: results of a randomized, double-blind, controlled trial investigating three dosages of cerebrolysin. Eur J Neurol. 2011;18: 59–68.
• Alvarez-Sabín J, Román GC. Citicoline in vascular cognitive impair-ment and vascular dementia after stroke. Stroke. 2011;42(Suppl 1): S40–S43.
• Cagnon L, Braissant O. Hyperammonemia-induced toxicity for the devel-oping central nervous system. Brain Res Rev. 2007;56:183–197.
• Chepkova AN, Sergeeva OA, Haas HL. Taurine rescues hippocampal long-term potentiation from ammonia-induced impairment. Neurobiol Dis. 2006 Sep;23(3):512-21.
• Fesenko UA. Piracetam improves children’s memory after general anaesthesia. Anestezjol Intens Ter. 2009;41:16–21. Polish
• García-Cobos R, Frank-García A, Gutiérrez-Fernández M, Díez-Tejedor E. Citicoline, use in cognitive decline: vascular and degenerative. J Neurol Sci. 2010;299:188–192.
• Guekht AB, Moessler H, Novak PH, Gusev EI; Cerebrolysin Investigators. Cerebrolysin in vascular dementia: improvement of clinical outcome in a randomized, double-blind, placebo-controlled multicenter trial. J Stroke Cerebrovasc Dis. 2011;20:310–318. 
• Holecek M. Side Effects of Long-term Glutamine Supplementation. JPEN J Parenter Enteral Nutr. 2012 Sep 18.
• Holinski S, Claus B, Alaaraj N, et al. Cerebroprotective effect of piracetam in patients undergoing coronary bypass surgery. Med Sci Monit. 2008;14:153–15.
• Kanamori K, Ross BD, Chung JC, Kuo EL. Severity of hyperammonemic encephalopathy correlates with brain ammonia level and saturation of glutamine synthetase in vivo. J Neurochem. 1996 Oct;67(4):1584-94.
• Lemberg A, Fernández MA. Hepatic encephalopathy, ammonia, glutamate, glutamine and oxidative stress. Ann Hepatol. 2009 Apr-Jun;8(2):95-102.
• Liu J. The effects and mechanisms of mitochondrial nutrient alpha-lipoic acid on improving age-associated mitochondrial and cognitive dysfunction: an overview. Neurochem Res. 2008;33:194–203.
• Mancuso C, Bates TE, Butterfield DA, et al. Natural antioxidants in Alzheimer’s disease. Expert Opin Investig Drugs. 2007;16:1921–1931.
• Malaguarnera M, Gargante MP, Cristaldi E, et al. Acetyl L-carnitine (ALC) treatment in elderly patients with fatigue. Arch Gerontol Geriatr. 2008;46:181–19
• Malaguarnera M, Gargante MP, Cristaldi E, et al. Acetyl-L-carnitine treatment in minimal hepatic encephalopathy. Dig Dis Sci. 2008;53: 3018–3025
• Malykh AG, Sadaie MR. Piracetam and piracetam-like drugs: from basic science to novel clinical applications to CNS disorders. Drugs. 2010;70:287–31
• Pantoni L. Treatment of vascular dementia: evidence from trials with non-cholinergic drugs. J Neurol Sci. 2004;226:67–70
• Parnetti L, Mignini F, Tomassoni D, Traini E, Amenta F.  Cholinergic precursors in the treatment of cognitive impairment of vascular origin: ineffective approaches or need for re-evaluation? J Neurol Sci. 2007;257:264–269.
• Ramalho SA, Nigam N, Oliveira GB, Alves de Oliveira P, Matos Silva TO, Passos dos Santos AG, Narain N. Effect of infusion time on phenolic compounds and caffeine content in black tea  Food Research International; 13 December 2012 [ahead of print]
• Secades JJ. Citicoline: pharmacological and clinical review. Rev Neurol. 2010;52 Suppl 2:S1–S62.
• Vale S. Current management of the cognitive dysfunction in Parkinson’s disease: how far have we come? Exp Biol Med (Maywood). 2008;233:941–951.
• Vince AJ, Burridge SM. Ammonia production by intestinal bacteria: the effects of lactose, lactulose and glucose. J Med Microbiol. 1980 May;13(2):177-91.
• Yurko-Mauro K. Cognitive and cardiovascular benefits of docosahexaenoic acid in aging and cognitive decline. Curr Alzheimer Res. 2010;7:190–196.

Choline Maximizes Carnitine Retention & Effects. Together, These Supplements May Promote Fat Loss + Lean Mass Gains and Boost Both Metabolic & Brain Health

The more choline the better the carnitine retention; and the better the retention the less high carnitine red mead you'll have to eat ;-)

If you are one of the many people who frequent boydbuilding-oriented bulletin boards, it is likely that you will have heard about the beneficial effects of exogenous insulin on the tissue storage of supplemental l-carnitine. When you hear something like this, or, as in this case, read about it in a forum, you can easily get the impression that it's only a matter of how much insulin you inject to turn an ergogenic aid, of which the majority of the scientific papers say that it is of questionable value, into a fat burning high performance powerhouse.

If said bulletin boards are not the only thing you frequent, i.e. if you are a regular at the SuppVersity, as well, you will probably also be aware of the fact that I strongly discourage the use of insulin in non-diabetic individuals - especially in cases as the one at hand, where it appears as if there were non-pharmacological alternatives to achieve increases in carnitine retention.

Using choline instead of insulin to maximize carnitine retention

I guess I could start this paragraph by stating that "it is a wonder that nobody appears to know about the interactions between choline and carnitine", but honestly, I have long given up to wonder when the respective information has to be looked up in studies that are older than 2-3 years. Studies as the one by Dodson and Sachan that was conducted at the University of Tennessee in the mid 1990 and the results of which werr published in the American Journal of Clinical Nutrition in 1996.

In the corresponding paper, Dodson and Sachan report the results of two experiments that were conducted to determine the effects of supplementary choline and/or pantothenate on the carnitine and lipid status of free-living humans:

Carnitine as Repartitioning Agent? + 7% Improvement in Lean- to Total Mass Ratio W/ HED of 1-1.5 of Carnitine/Day | more

"In experiment 1, adults receiving 13.5 mmol choline plus 1.4 mmol pantothenate/d had a significant decline in urinary carnitine excretion and renal clearance with nonesterfied carnitine (NEC) declining the most dramatically, 84%.

Additionally, serum NEC and total carnitine concentrations decreased significantly. No changes were observed in any of the serum lipids examined." (Dodson. 1996)

In the follow up experiment, Dodson and Sachan were able to show that the beneficial effects choline had on the retention of supplemental carnitine were not influenced by the provision of panthothenate, which had previously been used as a complement to carnitine and choline in clinical trials for a variety of neurologic and hyperlipidemic conditions.

Choline doesn't work for supplemental carnitine, only

Most of you will know that carnitine is not just a supplement, but also a non-essential nutrient. As long as we consume large enough quantities of lysine and methionine, the building blocks our bodies use to produce carnitine "on demand" we don't really need supplemental and probably not even dietary carnitine sources to survive (Mitchell. Feb. 1978).

Table 1: Total carnitine content of various foods (Mitchell. 1978)

Did you know that l-carnitine has for quite some time been touted to be a vitamin? The name vitamin BT is actually still sometimes used to refer to beta-hydroxy-y-trimethylamino butyrate aka carnitine; and that despite the fact that it can synthesized endogenously in mammals. It is thus not surprising that most carnitine-rich foods are animal products (see Table 1).

In view of our ability to produce "all the carnitine we need", it's actually not surprising that a sedentary slough is not going to benefit from respective supplements. And still, I had my reasonst to put the "all the canritine we need" in quotation marks. There are in fact situations, where research suggests that a lack of carnitine can become the bottle neck to fatty acid oxidization. The interesting thing about choline is now that it does not make a difference between endogenous and exogenous carnitine: Both are retained more effectively with an adequate / increased choline supply (Tsai. 1974, 1975; Carter. 1978; Hoppel). Low choline intakes, on the other hand, have been found to increase the urinary carnitine concentration and deplete the tissue stores independent of the overall nutritional status of the lab animals (Sheard. 1994).

In other words, the equation "higher choline = better carnitine levels" is valid irrespective of whether you are supplementing with beta-hydroxy-y-trimethylamino butyrate or simply producing it yourself.

SuppVersity Suggested Read: "Carnitine Loading Revisited: 3g Carnitine per Day Ward Off Vitargo Induced Fat Gain by Increasing Fatty Acid Oxidation and Total Energy Expenditure in 12 Week Human Study" | read more

Apropos nutritional status: I guess it's worth mentioning that fasting has been shown to lead to an increased mobilization of carnitine in order to facilitate the necessary increase of the carnitine-driven influx of fatty acids into the mitochondria in the absence of which the lean and obese subjects in a 1980 study by Hoppel et al. would have been starving.

Against that background it does not come as a surprise that the carnitine demands increase, when you go on a low carb or even ketogenic diet. In fact, Balaban-Gil et al. mention diet-induced carnitine deficiency as a potentially hepatoxic (=liver-damaging) side effect of long(er)-term ketogenic dieting (Balaban-Gil. 1998). They do yet also point out that the beginning liver damage can be sent into remission by the timely provision of supplemental l-carnitine.

On a side note: You do remember having read about the beneficial effects choline has on liver, did you? If not, I'd suggest you review my previous article "Choline: Stronger, Faster, Leaner & More Muscular, or Just Another Dumb-and-Barbell Story?" | read more.

More than just a life-insurance for the livers of keto dieters

As a diligent SuppVersity reader you will probably already know that choline, which used to be a staple supplement in the earlier days of bodybuilding, is a potent synergist to caffeine and carnitine - so potent in fact that I already devoted a whole post to the fat burning magic of the "CCC Stack" (read more).

Suggested Read:"Forgotten Dieting Aids: Choline, Carnitine, Caffeine and the Anti-Weight-Loss Plateau Effects of Sugar and Phosphates" | learn more

It would be pointless to repeat the discussion of the results of the 2003 study by Hongu and Sachan, so I'd suggest you (re-)read the corresponding article from February 2012, if you are interested how this combination lead to effortless weight loss in 19 healthy non-obese women.

I'd rather take a parting look at what actually happens to the carnitine in the Dodson and Sachan study. It was neither excreted nor pooling up in the blood of the 29 healthy volunteers when they consumed 13.5mmol of choline (ca. 1.4g of choline) as choline bitartrate (~3g) for 7 days and choline + of carnitine for additional three days.

Due to the fact that Dodson and Sachan who have been working at the University of Tennessee, back in the day, did not conduct muscle, let alone heart, brain, kidney and liver biopsies. We will (unfortunately) have to content ourselves with the results of a 1998 guinea pig  study from the same work group (Daily. 1998), if we want to understand the fate of the "missing" carnitine.

Total ingested (full bars) and absorbed (blue part of the bar) amount of dietary carnitine in mg/kg body weight (from Amino Acids for Super Humans)

When and how do you take carnitine and choline? If we put some faith into the non-published data from previous experiments by Dodson & Sachan, the responses should identical irrespective of whether you take the supps at one time or in several doses throughout the day. If we do now take the study by Hongu and Sachan as a guide the "recommended dosage" would amount to ~3g of choline bitartrate + 1.4g of carnitine - in view of the fact that Hongu's & Sachan's subject were women maybe up to 4g choline + 2g of carnitine per day for the heavier guys out there.

I know an animal model is not ideal, but it's better than trying to guess what caused the slight decline in serum levels and significantly lower urinary losses in the human trial from 1996 (Dodson. 1996); and the interaction of choline and carnitine in guinea pigs is more akin to humans than that of rodents, which is, according to Daily et al. (1998), probably a direct consequence of a lower choline oxidase activity in Guinea pigs vs. rats / mice.

What exactly happens to the carnitine, when you consume extra choline?

The study protocol Daily et al. used was slightly different from the one Dodson and Sachan had used in their 1996 human trial: Instead of carnitine and choline, the guinea pigs received only choline (+200% more than the regular diet would offer).

Figure 1: Changes in carnitine content of brain, liver, heart, kindney and muscle (left) and differences in body composition (right) extra choline vs. normal chow (Daily. 1998)

As you can see in Figure 1, there was a significant reduction in kidney carnitine (remember that this is where the carnitine is extreted) and concomittant increases in brain (specifically acetyl-carnitines) and muscle carnitine (total carnitine) content. The levels in the livers and hearts of the guinea pigs, on the other hand, remained stable (p > 0.05).

The question whether these changes were the cause or just correlates to the beneficial effects the provision of additional choline had on the body composition (see Figure 1, right) of the hairy mini remains to be answered, though. On the other hand, it is quite certain that the observations Daily et al. made in their lab animals stand in line with the results of Hongu et al.'s human trial from 2003, which clearly suggests that the "selective accretion of nitrogen and depletion of fat", of which Daily et al. state that it is an "important and unique consequence of choline–carnitine interactions" is not species dependent and occurs even in the absence of caffeine, the CNS stimulant Hongu et al. added to the equation.

Is choline the missing link, or rather the missing synergist due to which most of the studies that investigated the ergogenic effects of l-carnitine yielded very disappointing results?

In view of the fact that reviews as the one by Bass regularly refer to the fact that "muscle carnitine content is not easily increased with carnitine supplementation" (Brass. 2004) as one of the underlying reasons for the disappointing results that have been reported in a whole host of studies, it appears not too far-fetched to hypothesize that some of the trials may have yielded very different results if the researchers had used a combination of carnitine and choline.

Speaking of the synergy of choline + carnitine: You probably remember the study that claimed to show that the "high" carnitine content of red meat was carcinogenic, right? I have discussed the fallacy of this assumption in a separate article (read more) and I am not really interested to revive this discussion, that the moment.

What I am interested in, though, is red meat or rather the fact that the major carnitine sources in our diet, i.e. animal products, always come with choline. For beef, for example, the ratio is almost 1:1 (depending on the reference you use). At this point I could probably say something about "nature knows" best. That would be an understatement, though. If we are honest with ourselves, it's after all more like "nature knows" and "we are groping in the dark" - wouldn't you agree?

Bottom line summary: Before I summarize what you could have learned today if you actually read the article, I briefly want to remind you of the general health & performance benefits of choline I wrote about before (read more). After reading today's article you can thus expand the said list or your own mental notes about the useful effects of choline by the following items::

• The provision of choline reduces the high urinary excretion of carnitine which has always been the bottleneck of carnitine supplementation.
• In human studies, the combination of choline + carnitine + caffeine has been shown to promote fat loss. If we take the corresponding study as a reference, an effective dosing regimen would contain ~3g of choline bitartrate + 1.4g of carnitine.
• From animal trials we know that the changes in body composition are accompanied by significant increases skeletal muscle and brain total and acyl-carnitine levels. In view of the physiological role carnitine plays in the oxidation of fatty acids in the mitochondria, it is likely that the increase in muscular carnitine levels is mechanistically involved in the repartitioning effects (increased muscle decreased fat mass).
• The increased mobilization and clearance of carnitine in the fasted state suggest that choline and carnitine will be most useful, when you are dieting - in  this case even in the absence of additional caffeine, which is otherwise needed to increase the serum levels of readily oxidizable free fatty acids.
• Case reports from children on ketogenic diets suggest that l-carnitine supplementation can become mandatory to avoid liver damage, when patients are in full ketosis for a long period of time. Even in the absence of specific trials, it is thus reasonable to assume that a combination of l-carnitine and choline should have beneficial effects for anyone following a ketogenic or very low carb diet.

One last note: If you take a look at the interaction between choline and carnitine and the role carnitine plays in liver (=help oxidize fatty acids before they start clogging the liver → NAFLD), brain (=improve + maintain cellular energy status & neuronal function) and muscle (=improve fatty acid oxidation), it cannot be excluded that many of the previously listed health & perfomance effects of choline are actually mediated by its interaction with endogenous carnitine.

References:

• Ballaban-Gil K, Callahan C, O'Dell C, Pappo M, Moshé S, Shinnar S. Complications of the ketogenic diet. Epilepsia. 1998 Jul;39(7):744-8.
• Brass EP. Carnitine and sports medicine: use or abuse? Ann N Y Acad Sci. 2004 Nov;1033:67-78. Review.
• Carter AL, Frenkel R. The relationship of choline and carnitine in the choline deficient rat. J Nutr l978;108:l748-54.
• Daily JW III, Hongu N, Mynatt RL, Sachan DS.  Choline supplementation increases tissue concentrations of carnitine and lowers body fat in guinea pigs. The Journal of Nutritional Biochemistry. 1998; 9(8): 464–470.
• Dodson WL, Sachan DS. Choline supplementation reduces urinary carnitine excretion in humans. Am J Clin Nutr. 1996 Jun;63(6):904-10. 
• 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.
• Hoppel CL, Genuth SM. Carnitine metabolism in normal-weight and obese human subjects during fasting. Am J Physiol. 1980 May;238(5):E409-15.
• Mitchell ME. Carnitine metabolism in human subjects. I. Normal metabolism. Am J Clin Nutr. 1978 Feb;31(2):293-306. Review.
• Mitchell ME. Carnitine metabolism in human subjects. II. Values of carnitine in biological fluids and tissues of "normal" subjects. Am J Clin Nutr. 1978 Mar;31(3):481-91. Review.
• Sheard NF, Krasin B. Restricting food intake does not exacerbate the effects of a choline-deficient diet on tissue carnitine concentrations in rats. J Nutr. 1994 May;124(5):738-43.
• Tsai AC, Romsos DR. Leveille GA. Significance of dietary carnitine for growth and carnitine turnover in rats. J Nutr l974;104:782-92.
• Tsai AC, Romsos DR. Leveille GA. Determination of carnitine turn over in choline-deficient and cold-exposed rats. J Nutr 1975;105: 301-7. 

Particle Size & LCAT Analysis Shows: Three Whole Eggs per Day Improve Lipid Profile in Men & Women W/ MetSyn. Plus: Up to 700% Increased Lipid Oxidation in Hardboiled vs. Fresh Omega-3 Eggs From Hens on Fish Oil Diet

"Not the yellow part of the egg!", is the literal translation of "Nicht das Gelbe vom Ei!", which is German and means "not exactly brilliant" - telling isn't it?

As a regular here at the SuppVersity you will probably have come across a line like "make sure to get at least 20g of quality protein (meats, poultry, eggs, fish, dairy) with every meal!" at least once. If we discard the fish, which is, outside of the occasional discussions about mercury and other heavy metals in our food chain, about the only of these highly nourishing foods, the various experts appear to agree is healthy for you, all of them are to be consumed only "on occasion", "in moderation" and preferably in their "lean" or "low-fat" varieties. And while you (should ;-) have read about the beneficial effects of full-fat dairy, yesterday, and about the actually not so bad "bad meats" in the "Meat-O-Logy" post from August 17, 2012, I thought I'd take the forthcoming publication of a paper by Christopher N. Blesso, Jeff Volek et al. as incentive to remind you not to make the mistake and flush the yolk, the best part of the egg, down the toilette.

Recipe for disaster? Take 20 men and women with metabolic syndrome, 5040 whole eggs...

With 40 men and women aged 30–70 years who had been classified with metabolic syndrome the researchers from the University of Connecticut and University of Antioquia in Medellin, Columbia, intentionally picked a representative sample from the ever-increasing number of people with metabolic syndrome for their 12-week dietary intervention trial. I mean, who if not the men and women in this high risk group should suffer from the negative side effects of the cholesterol-laden yellowy, orange-yellow heart killers in disguise?

AHA Definition of Metabolic syndrome

according to Grundy. 2004

Abdominal obesity (waist)
Men	>102 cm
Women	>88 cm
Triglycerides	≥150 mg/dL
HDL cholesterol
Men	40 mg/dL
Women	50 mg/dL
Blood pressure	≥130/≥85
Fasting glucose	≥110 mg/dL

The assumption that subjects with established metabolic syndrome (for the "official" criteria check out the table on the right) is however one of the very few things the study at hand and studies like the one that triggered such an upheaval in August (see Sciencedaily. 2012) have in common. Instead of relying on total, low density and high density lipoprotein levels in the sera of their study participants, Blesso et al. conducted a differential analysis that included paricle number, size (measured by nuclear magnetic resonance spectroscopy), apolipoproteins (apos), oxidized LDL (oxLDL), cholesteryl ester transfer protein (CETP) and lecithin-cholesterol acyltransferase (LCAT) activities at baseline and after 12-weeks on a carbohydrate-restricted diet (25%–30% energy) that contained either  3 regular whole eggs/day (EGG, n = 20) or the volume equivalent in form of a yolk-free egg substitute (manufactured by Sysco Corporation, Houston, TX).

Accordingly, the subjects in the "real egg group" (EGG) consumed approximately 534 mg cholesterol, 0 g carbohydrate, 16 g protein, 12 g fat per day (186 kcal) from hole eggs, while a single serving of the cholesterol and fat-free substitute (SUB) contained approximately 2 g carbohydrate, 14 g protein, no fat, no cholesterol and only 60 kcal.

To minimize possible confounding factors and allow for "blinding" (you can't tell me you don't taste the difference between egg whites and whole eggs, but alas...), the eggs / egg supplements were prepared in advance.

"Compliance was monitored by use of weekly questionnaires and collection of empty product containers. Participants were asked to maintain their normal physical activity, medications, and dietary supplement usage upon starting the 12-week study." (Blesso. 2012)

To be able to track the dietary intake of the subjects, all participants had to fill out 5-day dietary intake records (3 weekdays + 2 weekend days) at baseline, week 6 and week 12 of the study period. Based on the analysis of the respective data, the scientists determined that

• increased relative protein intake from 17.3%± 3.0% to 23.9%± 4.1%
• increased relative fat intake from 38.6%± 6.4% to 45.7%± 7.4%

• reduced total energy intake of -24% for all participants
• reduced relative CHO intake from 40.9 ± 7.4 to 28.3 ± 9.5% of total energy 
• reduced absolute CHO intake from 211.9 ± 51.8 to 114.5 ± 55.0 g/d 

Moreover, there were "significant differences between groups for dietary cholesterol intake (EGG vs. SUB, P < 0.0001) and dietary choline intake (EGG vs. SUB, P < 0.0001)"

• Δ cholesterol intake +106% in EGG vs. -38% in SUB
• Δ choline intake +52% in EGG vs. -12% in SUB

In that, it is important to point out that these changes and their metabolic effect should not be seen in isolation, because choline plays a major role in the metabolism, transport and incorporation of cholesterol into the cell membrane (read more about choline in "Old School Supplements - Choline: Stronger, Faster, Leaner & More Muscular, or Just Another Dumb-and-Barbell Story?").

"So, the men and women in the egg-group did not die?"

Not exactly, no. Rather than being as bad as cigarettes (as the aforementioned "study" that was published roughly 2 months ago would make us believe; Spence. 2012), the consumption of three eggs per day lead to rather favorable changes in the lipoprofile of the EGG consumers:

Figure 1: Lipoprotein serum levels (left), particle size and ratio of LDL to HDL count (right) of the 37 participants who completed the 12-week trial (data expressed relative to baseline; calculated based on Blesso. 2012)

Contrary to the lipoprotein metabolism, which showed differential (and general positive) effects in response to whole egg consumption, the reductions in circulating insulin, HOMA-IR and triglycerides did not show statistical significant inter-group differences:

triglycerides: -29% vs. -18%
• insulin: -21% vs. -14%
• HOMA-IR: -22% vs. -18%

The existing changes were yet on average still somewhat more pronounced in the EGG group, which is an observation you could interpret as joint effect of low carb dieting + egg eating.

The same goes for the increase in LCAT activity the researchers observed only in the EGG group. Since LCAT is critically important in facilitating HDL particle stability and HDL maturation, its elevation in the EGG group

"[...] could be indicative of an enhanced capacity for HDL maturation and may help explain the shift towards larger HDL particles seen with egg feeding" (Blesso. 2012; my emphasis)

and would thus favor enhanced HDL-mediated reverse cholesterol transport from (e.g. the endothelium of your coronary heart arteries) back to the liver.

What about the lipid oxidation?

If we assume that part of the negative effects that have been ascribed to the consumption of egg(-yolks) are brought about by oxidized fatty acids and respective byproducts, it appears wise not to hard-boil your eggs, because the prolonged heat exposure during hard-boiling increases the oxidation of fatty acids. Aside from temperature and duration of the heat exposure, Cortinas et al. also identified the fatty acid composition of the eggs and their vitamin E content as crucial factors that will influence the formation of oxidized lipids. Since the fatty acid composition of the eggs depends on the diet the hens are fed, you would be particularly ill-advised to hard-boil eggs from hens who were fed a diet enriched in fish oil.

As you can see in the figure above the "fish oil eggs" with their readily oxidized long-chain PUFAs had by far the highest  TBA values (expressed as ng MDA/g dry matter) - specifically, if the hens did not receive additional vitamin E that would be transfered to the eggs and reduce the heat-induced oxidation of the long-chain PUFAs in the "omega-3 eggs" during cooking.

So what's the verdict? In conjunction with the aforementioned additive effects on glucose management and the slightly more pronounced decrease in oxidized LDL (both are probably rather a result of the -12% reduction in carbohydrate content of the participants' diets than the effect of the eggs), the statistically significant improvements in LCAT activity in the EGG eater group speak in favor of the notion that three whole eggs a day will still help you keep the doctor away. Whether they will suffice to throw his statins away, as well (see "Eat Whole Eggs All Day and Throw Your Statins Away?"), will yet have to be elucidated and appears overall questionable - and this goes regardless of whether you belong or don't belong to the relatively small subgroup of people who actually benefit from taking a statin, or not.

What stands out of question, however, is that the "side effects" of three egg yolks per day are quite distinct from those of cigarettes, the consumption of which the headline of the aforementioned ScienceDaily.com article implicitly equates with "egg yolk consumption". And just like smoking and eating eggs are two very different pairs of shoes, there is a huge difference between this well-controlled trial, on the one hand, and the undifferentiated observational hokum that got Spence et al. so much media attention back in August, when they concluded "Our findings suggest that regular consumption of egg yolk should be avoided by persons at risk of cardiovascular disease." (Spence. 2012), on the other hand.

References:

• Blesso CN, Andersen CJ, Barona J, Volek JS, Fernandez ML. Whole egg consumption improves lipoprotein profiles and insulin sensitivity to a greater extent than yolk-free egg substitute in individuals with metabolic syndrome. Metabolism. 2012 Sep 26.
• Cortinas L, Galobart J, Barroeta AC, Baucells MD, Grashorn MA. Change in α-tocopherol contents, lipid oxidation and fatty acid profile in eggs enriched with linolenic acid or very long-chain ω3 polyunsaturated fatty acids after different processing methods. J. Sci. Food Agric. 2003; 83: 820–829.
• Grundy SM, Brewer HB Jr, Cleeman JI, Smith SC Jr, Lenfant C; American Heart Association; National Heart, Lung, and Blood Institute. Definition of metabolic syndrome: Report of the National Heart, Lung, and Blood Institute/American Heart Association conference on scientific issues related to definition. Circulation. 2004 Jan 27;109(3):433-8.
• ScienceDaily. Egg yolk consumption almost as bad as smoking when it comes to atherosclerosis, study suggests. August 13, 2012 < http://www.sciencedaily.com­ /releases/2012/08/120813155640.htm > accessed on October 03, 2012.
• Spence JD, Jenkins DJ, Davignon J. Egg yolk consumption and carotid plaque. Atherosclerosis. 2012 Oct;224(2):469-73.

Choline Supplementation Accelerates Fat Loss During Crash Diet in Female Athletes: 2g Choline Double the Rate of Fat Loss in the Last Week Before the Competition

Can you hit the fat hard with choline?

I want to be honest with you. I am a huge fan of choline and truly believe that it is hugely under-appreciated, but the prominent relative (not absolute) increase in body fat loss in study at hand must be interpreted with caution - no matter how statistically significant the "choline advantage" may be.

Before we can get to said "cautious interpretation", let's briefly take a look at what exactly Gehan Elsawy, Osama Abdelrahman, and Amr Hamza from the Zagazig University and the Mansoura University in Egypt did to produce a 100% increase in body fat loss in their 22 female study participants (15 taekwondo and 7 judo athletes).

You can learn more about choline at the SuppVersity

Huge GH Spikes from GPC!?

Choline & Classic Bodybuilding

Effects of Choline Deficiency

Caffeine, Choline, L-Car = Fat Loss

Choline Maximizes Carnitine Storage

Choline as a Nootropic?

The idea was to clarify the magnitude of rapid body mass reduction among Egyptian judokas, in order to identify the scientific basis and justification for such practices. In that, the researchers were particularly interested in the effects of choline supplementation on bodymass reduction and leptin levels among their females taekwondo and judo athletes.

The athletes were divided into two groups, according to their body mass; the experimental group contained ten female athletes, and the control group twelve female athletes. At the time of enrollment, all the subjects were healthy, according to a medical information questionnaire, and none of the subjects had any specific dietary restrictions. Exclusion criteria included the use of any medication or supplement during the previous six months.

2.0g per day divided in two 1.0g doses of choline did the trick

For one week prior to a competition, the athletes in the experimental group took choline tablets (1.0 g) twice daily with a meal, equaling a total daily dose of 2.0 g (the scientists don't provide any information on the form of choline, they used, but their references suggest that it was PS, i.e. phosphatidylcholine). The control group received a placebo, and they participated in usual training (with 75% training intensity) at the same time as the choline group four times per week.

"According to Anni et al. (2011), choline supplementation appears to be safe and the authors recommend taking approximately 2.5 g one hour before a prolonged exercise session. The effective dose in sport studies is 0.2 g phosphatidylcholine 90% per kg of the body mass, which equals 2.1 g of choline for an 80-kg athlete. There is no requirement for a loading or maintenance phase and choline supplementation up to one hour before exercise has been shown to be effective in reducing fatigue." (Elsawy. 2014)

There was no standardized diet, there were no diet logs and there was no recording of training intensity and volume.

Body impedance a major source of error: The body fat analyses were conducted with Tanita Bioimpedance BC-418 devices. A technique that has only recently been shown to be highly sensitive to changes in body water (Slater. 2014) - changes as they occur regularly in female subjects and changes which could be influenced by the consumption of choline. I mean, generations of bodybuilders have popped choline tablets in an effort to reduce the subcutaneous water and get that cut dry look, judges want to see on stage (learn more).

Things that were assessed are body weight, body fat (see red box above), serum and urinary choline, as well as back and leg strength.

Figure 1: Changes in leptin, plasma choline, body fat (%), BMI, leg & back strength within the last week of precompetition dieting with or without the addition of 2g of choline (undisclosed form) in a recent study by Elsawy et al. (2014).

Statistically significant differences were observed for plasma choline (obviously), leptin and the change in body fat (-1% vs. -2% in the choline group). It would be nice if we also knew if this affected the food and/or water intake and/or if we had confirmation from DEXA and caliper data that the body fat difference was more than just body impedance b*s* - unfortunately, none of these data are available.

Choline could also boost fat loss by boosting carnitine levels | more

Bottom line: Now, if it was not for the difference in leptin, the study probably wouldn't have made it into the SuppVersity news - a 1% difference, in body fat (%), as statistically significant as it may be, is after all hardly worth mentioning, when it was measured by body impedance in a tranining phase where hypohydration often beomes an issue. In conjunction with the reduced MDA levels, a clear sign of significant anti-oxidant effects of choline, and the general role of choline in the metabolism of fat (Hanin. 1987), which has also been linked to a greater level of satiety (Wurtman. 1977), I am yet willing to say: Adding 2.0g of choline in form of cheap choline bitartrate is certainly worth a try - I mean, what to you have to lose aside from some of the money you would otherwise spend on other unproven fat burners? And yes, I am pretty sure that in case it does work, it will work for both: women, as in the study at hand, and men.

References:

• Elsawy et al. "Effect of Choline Supplementation on Rapid Weight Loss and Biochemical Variables Among Female Taekwondo and Judo Athletes." Journal of Human Kinetics volume 40/2014, 77-82.
• Hanin I, Ansell GB. "Lecithin: Technological, Biological, and Therapeutic Aspects". Plenum Press, NY, 180-181; 1987.
• Slater, Gary. "Assessing Body Composition of Athletes." Sports Nutrition for Paralympic Athletes (2014): 189.
• Wurtman RJ, Hirsch MJ, Growdon JH. "Lecithin consumption raises serum-free-choline levels." Lancet, 1977; 2: 68-69

Amino Acids for Super Humans. Part IV: Purported Ergogenics - Beta Alanine, Carnitine, Glutamine

This relevant for all of you - regardless of whether you can or cannot spare the time: Tune in live and listen to me @ Carl Lenore's Super Human Radio to learn that amino acids are far more than just the building blocks of the proteins of your body...

Listen live to SHR @ 12:00PM ET

Amino Acids for Super Humans. Part IV
Purported Ergogenics - Beta Alanine, Carnitine, Glutamine

In the fourth installment of the show, we are going to look at three of those amino acids everyone interested in fitness and body building knows: Beta Alanine, Carnitine and Glutamine. Will the increase performance, fat loss and muscle gains, as some supplement companies would have it, or will they mostly cost your hard earned money or even make you sick? Tune in live @ 1pm ET and learn more!

19x Increase in Growth Hormone 60min After Ingestion of 1g of Glycerophosphocholine (GPC) in Young Male Subjects

Image 1: Don't worry one thing is sure - GPC won't give you a gut like that ;-)

A couple of days ago, I have written about the exorbitant choline consumption of some of bodybuilding legends, Randy Roach writes about in his Muscle Smoke & Mirrors Vol. II  and to be honest, I was quite surprised that among the many responses I got, none broached the issue of the latest and greatest supplemental choline spin-off: Glycerophosphocholine, or short GPC - a supposedly superior, because fat-bound (hence "glycero" as in "glycerol) and highly bioavailable bean-derived form of choline. Whatever the reasons may be, I suppose you still won't mind to hear that a recent experiment that was conducted at the Faculty of Sport and Health Science of Ritsumeikan University in Shiga, Japan, yielded quite interesting results with respect to the endocrine and metabolic short-term effects of 1,000mg of GPC in 8 healthy male (25+/1y) subjects (Kawamura. 2012).

19x more growth hormone 60 min after the ingestion of 1,000 mg GPC, ...

And no, the subheading above does not contain a typo, at least not in the number, which is "nineteen" as in +1800%, which was, as the data in figure 1 shows, the average increase in serum growth hormone concentration exactly 60 minutes the lean subjects (11% body fat) experienced after ingestion of the GPC caps compared to placebo (the study was double-blinded and randomized, there was a period of two weeks in-between the testing days).

Figure 1: Fatty acid metabolism (left), choline levels (middle) and growth hormone levels (right)  6min0 and 120 min after the ingestion of placebo pills or 1,000mg glycerophosphocholine (Kawamura. 2012)

It is unquestionable the data in figure 1  looks pretty impressive. Yet not all changes are in fact statistical significant (I marked them by adding the relative differences between active and placebo arm) and as exciting a growth hormone spike of +1800% may sound, the subsequent drop to levels below the placebo arm should remind you, a seasoned SuppVersity veteran of the futility of supplemental "growth hormone secretagogues", such as arginine, leucine, a combination of both etc.; they all share a fundamental weakness - after each spike there is a huge trough.

...but just  6.88x more GH production over 2h

And therefore it should not really surprise you that we have the 2-h AUC, i.e. the area under growth hormone curve, a measure for the total amount of growth hormone that is released in the course of the timespan for which the AUC was measured, was "only" 6.88x larger after the ingestion of the glycerophasphatecholine supplement, than after placebo(cf. figure 2).

Figure 2: 2h AUC for free choline, free fatty acid, 3-hydroxbutyrate (ketones) and growth hormone after the ingestion of 1,000mg GPC; data expressed relative to placebo control (Kawamura. 2012)

Still, even this obviously transient increase in growth hormone in the fasted that occured in response to the +38-51% increase in circulating plasma choline went hand in hand with profound increases in free fatty acids (due to the lipolytic effects of GH; cf. Marcus. 1994) and ketone bodies, which is a sign of increased fatty acid metabolism must be a good thing,... right? It would appear so, but without answering the following two fundamental questions "How does it work?" and "What does that mean", we will be having a hard time to justify this conclusion. After all, the ingestion of 4mg/kg caffeine have been shown to illicit a >500% increase in free fatty acids (FFA) after only 40min in trained subjects (LeBlanc. 1985) and still none of the caffeine laden "fat burners" with other ingredients, which further augment the lipolytic effects of caffeine will actively burn body fat.

So what's the mechanism of action? And what does it tell us about the real world implications?

The working principle Kawamura et al. suggest is actually quite straight forward: With increasing serum choline levels, the influx of choline into the brain will increase as well. This will augment the synthesis of acetylcholine, which, in turn, has been found to decrease the concentration of somatotropin release-inhibiting factors right at the hypothalamic level and thus disinhibit the production of growth hormone (Blusztajn. 1983). Put simply: Somewhere down the line the increase in serum choline will pull the breaks that keep your body from producing growth hormone.

A note to all the stim-junkies out there: I guess, you will be intrigued (or shocked?) to hear that blocking the catecholamine induced stimulation of the a2-adrenergic receptor with yohimbine has been shown to negate the aforementioned growth hormone promoting cascade. And that irrespective of whether you try to augment it by supplements or just want to keep your natural rhythm intact (Minamitani. 1989). The first real-world implication would thus be "don't take your GPC alongside alpha-2 antagonists such as yohimbine" (better not take those at all ;-)!

Now this raises the question does that matter? With arginine and lysine, we already know that it doesn't, but maybe we just have a larger effect size here? To answer the last question first - the spike is in fact spectacular and way above the average response to the long-touted GH boosters arginine, lysine or glutamine, which ranges from "no effect at all" (Carlson. 1989) over 4.5x (Welbourne. 1995) to the whopping 13x increase in response to an intravenous injection of  0.5 g arginine/kg (Tanaka. 1991). If do yet take another look figure 2 you will notice that I inserted a quote from Kawamura et al.'s discussion of the results into the graph - a quote that is of paramount importance to quantify the real world significance of these ostensibly HUGE increase in GH (which you will certainly see referenced by respective supplement manufacturers in their glossy marketing material, very soon):

"The GPC-induced increases in GH levels observed in this study were of a comparable degree to the increase induced by moderate-intensity exercise"

I guess, you don't need me to tell you how "effective" popping a couple of those pills is thus going to be compared to training and diet alone in furthering your muscle gains and fat loss. And in terms of overall and cardiovascular health, you should already know from my previous blogpost, "Old School Supplements: Choline  Faster, Stronger, Leaner & more Muscular" that regular dietary choline as in eggs, meats, fish, leafy greens, etc. will do just as fine.

How much choline do you need? According to Coates et al. plasma choline concentrations can double after a 2-egg meal (~225mg choline) by up to two-fold (Coates. 2005). That would effectively be more than what we see as peak increase in the study at hand. And certainly puts the "need" for supplemental choline into perspective. The LD50, i.e. the purportedly fatal dosis, after the ingestion of which 50% of the subjects would die, is "of the order of 200-400g" (Gilman. 1980) - an amount of choline your tummy probably would not hold o to long enough to be absorbed, anyway ;-)

And in the unfortunate case that you do believe that you are running short of choline, because you don't eat all the good choline containing foods out of ethical or whatever other reasons, and thus insist on supplementing, I suggest you yourself a 500g container of choline bitartrate powder (don't let that become wet, though! It will stink like rotten fish ;-). Those 500g of choline bitartrate (40% choline, 60% tartate) will cost you about as much as 60x300mg caps of the overpriced GPC and has been "scientifically proven" (not in supplement company terms, but in SuppVersity terms) to safely increase circulating and brain choline levels and its metabolites after oral ingestion, as well (Stoll. 1996; Babb. 2004). And let's be honest, even if the effects on growth hormone were GPC specific - even on the boards, people have meanwhile realized none of those arginine + lysine GH boosters does make a difference and not because they would not produce transient increases in GH, but simply because those are physiologically meaningless and mostly compensated for in the course of 24h.

References:

1. Babb SM, Ke Y, Lange N, Kaufman MJ, Renshaw PF, Cohen BM. Oral choline increases choline metabolites in human brain. Psychiatry Res. 2004 Jan 15;130(1):1-9.
2. Blusztajn JK, Wurtman RJ. Choline and cholinergic neurons. Science 1983;221:614–20.
3. Coates, P.M., Blackman, M.R., Cragg, G.M., Levine, M., Moss, J., White, J.D. (Ed), Encyclopedia of Dietary Supplements. Marcel Dekker, New York, NY. 2005. p. 108. . 
4. Gilman, A. G., L. S. Goodman, and A. Gilman. (eds.). Goodman and Gilman's The Pharmacological Basis of Therapeutics. 6th ed. New York: Macmillan Publishing Co. Inc. 1980. p. 1575. 
5. Carlson HE, Miglietta JT, Roginsky MS, Stegnik LD. Stimulation of pituitary hormone secretion by neurotransmitter amino acids in humans. Metabolism 1989;38:1179
6. Kawamura T, Okubo T, Sato K, Fujita S, Goto K, Hamaoka T, Iemitsu M. Glycerophosphocholine enhances growth hormone secretion and fat oxidation in young adults. Nutrition. 2012 Jun 5. 
7. LeBlanc J, Jobin M, Côté J, Samson P, Labrie A. Enhanced metabolic response to caffeine in exercise-trained human subjects. J Appl Physiol. 1985 Sep;59(3):832-7.  
8. Liu H, Bravata DM, Olkin I, Friedlander A, Liu V, Roberts B, Bendavid E, Saynina O, Salpeter SR, Garber AM, Hoffman AR. Systematic review: the effects of growth hormone on athletic performance. Ann Intern Med. 2008 May 20;148(10):747-58. Epub 2008 Mar 17.
9. Marcus C, Bolme P, Micha-Johansson G, Margery V, Brönnegård M. Growth hormone increases the lipolytic sensitivity for catecholamines in adipocytes from healthy adults. Life Sci.1994;54(18):1335-41.
10. Minamitani N, Chihara K, Kaji H, Kodama H, Kita T, Fujita T. Alpha 2-adrenergic control of growth hormone (GH) secretion in conscious male rabbits: involvement of endogenous GH-releasing factor and somatostatin. Endocrinology 1989;125:2839–45.
11. Stoll AL, Renshaw PF, De Micheli E, Wurtman R, Pillay SS, Cohen BM. Choline ingestion increases the resonance of choline-containing compounds in human brain: an in vivo proton magnetic resonance study. Biol Psychiatry. 1995 Feb 1;37(3):170-4. 
12. Welbourne TC. Increased plasma bicarbonate and growth hormone after an oral glutamine load. Am J Clin Nutr 1995;61:1058

Amino Acids for Super Humans. Part III - Sulfur, More Than Just Rotten Eggs.

This relevant for all of you - regardless of whether you can or cannot spare the time: Tune in live and listen to me @ Carl Lenore's Super Human Radio to learn that amino acids are far more than just the building blocks of the proteins of your body...

Listen live to SHR @ 12:00PM ET

Amino Acids for Super Humans. Part III
Sulfur, More Than Just Rotten Eggs
update: Episode available for download, now!

In the third installment of the show, we are going to look at the commonly overlooked. yet vitally important sulfur-containing amino acid methionine, its "children" and "grand children", cysteine, n-acetyl-cysteine (NAC) and the purported "cell volumizer" taurine.

As usual, I will do my best to provide relevant examples and relate the theory to practical advice. The magic of individual amino acids will be tackled in the shows to come.

update: Episode available for download, now!

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