Science Round Up Seconds: 30-60% More Testosterone w/ 2.5g D-Aspartic Acid in Fertility Trial and Nicotine Amplifies Cardiotoxic Effects of ECA. Plus: Data on DHEA & Estrogen & Breast Cancer, Fermented Teas, AMPK, AKT & Co

DAA is probably not going to hurt your heart, but it's more likely to father a child than to build those abs. Ephedrine & Caffeine on the other hand, could help you get there, but esp. if you are also smoking you are increasing the risk that the kids you fathered using DAA will soon be without their begetter.

I guess most of all will have listened to the podcast of yesterday's installment of the SuppVersity Science Round Up already. If you didn't you have been missing Carl and me discuss new on the pro-carcinogenic effects of aspartame, the never-ending story of the fattening artificial sweeteners, the benefits of oat beta-glucans for weightloss, -maintenance and gut health, the way sorghum proanthocyanidins can lower the GI of carbohydrates and make them less susceptible to enzymatic breakdown in the small intestine, and more.

Actually this more, i.e. the news on DHEA, its metabolits and their proliferative effect on breast cancer cells, as well as the information about the beneficial effects of fermented teas on blood glucose management, reminded me of the fact that as how like the SuppVersity Science Round Up is a very good place discuss and explain things, but not exactly the place to present detailed data. Therefore, I decided to prelude the Seconds by adding a couple of graphs which illustrate what has been said on the last show. Thus, you can look at the figures while listening to the podcast.

Supportive material for the DHEA and fermented tea news

For this first installment of the Seconds I did, you guessed it, pick the aformentioned news on DHEA and the different fermented teas (see figure 1) that are  based on studies by Miller (2012) and Yamashita  (2012), respectively.

Effect of 7 days of oolong tea, black tea, pu-erh tea, instead of water on Δglucose AUC (left), AMPK, AKT and PI3K expression in skeletal muscle  of male mice (Yamashita. 2012)

Effect of estradiol (E2), DHEA and its metabolits 7-OXO,  androstenediol, and androstenedione on breast cancer cell proliferation (based on Miller 2012)

So much for the additions to visuals for the podcast, let's get to the new stuff... or actually the seconds. Of course, the seconds ;-)

• How to brew your own sodium d-aspartic acid The best thing about this study actually is that the scientists disclose how you can easily make your own PH stable sodium-d-aspartate from the cheap stuff you buy at your favorite bulk supplier: Take 2.66 g of D-aspartic acid neutralize it with 0.46 g of NaOH in 10 ml distilled water and you get a final pH of 6.5 - 7-0 - that's it, you are good to go.
  New study on d-aspartic acid confirms - 30-60% increase in testosterone and LH in infertile men (D’Aniello. 2012) Despite the fact that this is a non-sponsored study by researchers from the Hospital “S. Luca” in Vallo della Lucania, Italy, I am about as 'unpsyched' about the data the scientists present, as I am about the real world results of d-aspartic acid (DAA) supplementation in young weight training men.
  
  It's already telling that D'Aniello et al. mention the increase in testosterone and luteinizing hormone (LH) only as an aside and consider it as a "save", or I guess you better say "tolerable" side effect of a treatment  that did effectively double the amount of D-aspartic acid in the seminal plasma and did thus (at least the scientists belive in a mechanism here) increase the fertility in both, patients with reduced sperm motility and sperm count, and those who suffered only from reduced motility.
  
  The actual 'success rate' in terms of pregnancy rates after 2-3 months of treatment with 2.66g/day of DAA per day was however not exactly really earth-shattering, either. Of the patients with both low sperm count and sperm motility (oligo-asthenozoospermia) 4% fathered a child; of those who suffered 'only' from a low sperm motility (asthenozoospermia) 33% eventually managed to become daddy.
  
  Without baseline testosterone levels, of which I would not be surprised if they had been rock bottom (both oligo-asthenozoospermia and asthenozoospermia usually go hand in hand with increased oxidation and that in turn is associated with low testosterone and suppressed LH levels), this study is however about as worthless in terms of the purported ergogenic effects of DAA, as all previous human trials. That said, you could obviously mix yourself the above concussion in case you and your significant other are planning to start a new or to expand your existing family in the near future. I guess, it's unlikely that it's going to hurt.
  
  Suggested read: All About the Role of Androgens & Co in Building Muscle
   
• Putting an "N" as in "nicotine" into "EC" amplifies the negative effects of ephedrine and caffeine on your heart and may well be the reason for many of the (few) deadly side effects that occurred in the day before the ban (Brown. 2012) When a group of researchers from the Arkansas State University tried to get to the bottom of the (in some cases) fatal cardiovascular side-effects, which were the main reason for the FDA to pull ephedra-containing supplements from the market, Christopher E. Brown and his colleagues observed ...

  "[...] a synergistic effect on the rat cardiac morphology [...] as a result of intera tions between nicotine, caffeine, and Ephedra. The cardiotoxicity caused by combination dosing of Ephedra and caffeine has already been shown; however, the present study revealed an enhancement of cardiotoxicity when nicotine was administered in combination with Ephedra and caffeine." (Brown. 2012)

  The scientists had exposed male Sprague-Dawley rats to (1) synthetic combinations of nicotine (0.2 mg/kg/day), ephedrine (0–30 mg/kg/day), and/or caffeine (0–24 mg/kg/day) as well as (2) an extract from a caffeine-containing Ephedra supplement (Metabolife 356). The relatively high dose treatments were administered for only 3 days either in the full or half dose and with and without nicotine pre-treatment to model the effects of different dosing regimen on smokers and non-smokers.
  
  

  

  Figure 1: Light micrograph of representative nuclear pro-files (background, red = atypical, green = normal nuclei; my emphasis) and volume (%) of atypical cardiac cells in anterior left ventricle of the rodents (Brown. 2012)

  As far as the results go, a a brief glance on the exemplary data in figure 1 should actually suffice to see, that a baseline "N" + "EC"  stack (as in any smoker who would take ephedrine + caffeine to lose weight or psyche himself up) could eventually pave the way to the emergency room.
  
  While the data from the anterior left ventricle and anterior interventricular septum (not shown) would suggest that the identically dosed synthetic versions of caffeine and ephedrine were slightly more detrimental, than the herbal supplement  in which the Ephedra came from a standardized Ma Huang extract and part of the caffeine from Guarana, this effect was not present in either the posterior left or the anterior right or posterior right ventricle (data not shown).
  
  Apropos interventricular septum (IVS), with increases in atypical cardiac cell volume of up to 1.5% in the anterior IVS even without nicotine pre-treatment, the stout wall that separates the lower chambers was most susceptible to the effects of caffeine and ephedrine:
  

  "In the anterior section of this region, both caffeine + ephedrine combination as well as the multicomponent supplement Metabolife 356 resulted in larger numbers of atypical cells compared to water controls, in both saline- and nicotine-pretreated rats. However, only rats pretreated with nicotine responded negatively to supplements in the posterior region of the IVS. " (Brown. 2012)

  If you consider the high-pressure forces it must sustain for proper ejection volume to the arterial vasculature it should be obvious that "these changes to the ventricular tissue could be particularly detrimental to overall cardiovascular health" (Brown. 2012). Bad news? Why? At least you do now have another good reason to stop smoking... what, oh yeah, I forgot: This is irrelevant because Ephedra has been banned anyway ;-)

While I do have a couple of other Seconds I am a bit pressed on time, today. Don't worry sooner or later they will appear ither on the SuppVersity Facebook Wall, where I am posting at least half a dozen of exclusive links and mini-items, comments and more you won't find on www.suppversity.com. So, I'd suggest you do now first listen to the podcast (if you have not already done so), then check out the latest SuppVersity Facebook News and when you are done with that wait till tomorrow for this weeks installment of On Short Notice.  

References

• Brown CE, Trauth SE, Grippo RS, Gurley BJ, Grippo AA. Combined Effects of Ephedrine-Containing Dietary Supplements, Caffeine, and Nicotine on Morphology and Ultrastructure of Rat Hearts. Journal of Caffeine Research. 2012; 2(3).
• D’Aniello G, Ronsini S, Notari T, et al. D-Aspartate, a Key Element for the Improvement of Sperm Quality. Advances in Sexual Medicine, 2012, 2, 47-53.
• Miller KKM, Al-Rayyan N, Ivanova MM, Mattingly KA, Ripp SL, Klinge CM, Prough RA. DHEA metabolites activate estrogen receptors alpha and beta. Sterespectivelyroids. November 01, 2012. Ahead of print.
• Yamashita Y, Wang L, Tinshun Z, Nakamura T, Ashida H. Fermented Tea Improves Glucose Intolerance in Mice by Enhancing Translocation of Glucose Transporter 4 in Skeletal Muscle. J Agric Food Chem. 2012 Nov 5.

When Hype Meets Reality: D-Aspartic Acid Turns Out to Be Another Supplemental Nonstarter in First Human Trial With Any Relevance for Healthy Young Men

Training "on" D-Aspartic Acid is like training on guar gum.

Busted! D-Aspartic acid aka DAA is another "natural anabolic agent" that turns out to be more of a revenue- than a hormone-booster with muscle building prowess. And you know what? It took Darryn S. Willoughby and Brian Leutholtz from the  Exercise and Biochemical Nutrition Lab, Human Performance, and Recreation at Baylor University exactly 28 days and 20 apparently healthy, recreationally active, resistance trained (3x per week or more in the past year) men with an average age of 22.8 ± 4.67 years (BMI 24.65 kg/m²) to prove that d-aspartic acid (DAA) supplements belong to this never-ending list of supplemental non-starters.

Things that work: 28 days, 4x per week heavy resistance training; things that don't work DAA capsules

Willoughby and Leutholtz assigned their participants in a random, double-blinded fashion to one of the two study arms. Subjects in both arms of the study followed a standardized periodized 28-day resistance training program split into 2 upper-extremity (A1, A2) and 2 lower-extremity (B1, B2) exercise sessions each week. With an overall training volume of 16 exercises sessions with 9x upper- and 8x lower-body exercises

• A1, A2 - upper body: bench press, lat pull, shoulder press, seated rows, shoulder shrugs, chest flies, biceps curl, triceps press down, and abdominal curls
• B1, B2 - lower body: leg press, or squat, back extension, step ups, leg curls, leg extension,heel raises, and abdominal crunches

the same workout hat shown to elicit significant improvements in body composition in two previous studies on VPX' preworkout products (learn more; cf. Shelmadine. 2009; Spillane. 2011). The participants performed 3 sets of 10 repetitions at 70% - 80% of the previously established 1-RM for all exercises and had 2 minutes of passive rest between the sets. Contrary to the said VPX studies, the participants had to refrain from taking any kitchen-sink pre-/post-workout supplements, as well as the obvious roids, prohormones or other ergogenic substances that would compromise the study results. Instead, they received either...

• 4 capsules containing 3 g of guar gum (PLA), or
• 4 capsules containing 3 g of D-ASP (Better Body Sports, Ventura, CA, USA)

The dosing was identical to the manufacturers recommendation and based on the previous study by Topo et al. (2009) that caused the whole DAA hype back ~5 years ago (see figure 2, as well).

Figure 1: Endocrine and body composition changes with placebo or DAA supplementation (Willoughby. 2013)

That fact the whole hoopla was exactly that: All hype! is difficult to refute, if you take a look at the study outcomes in figure 1 or the researchers' conclusion:

"[...] 28 days of D-ASP of supplementation at a daily dose of 3 g is ineffective in upregulating the activity of the HPG axis and has no preferential effects in which to increase skeletal muscle mass and strength in resistance-trained men." (Willoughby. 2013)

In their discussion of what might be the physiological reasons for the non-existent effects on the gonadal production of testosterone the researchers state:

Figure 2: Comparison of the hormonal effect of DAA in sedentary men with low testosterone levels (Topo. 2009) vs. trainees with normal to high levels of testosterone (Willoughby. 2013)

"Based on our data presented herein, this [=the fact that the serum levels of DAA were higher than normal, but nothing happened] may indicate another potential mechanistic reason why the HPG [hypothalamus > pituitary > gonads] axis was not affected by D-ASP supplementation. Although we observed nonsignificant increases in serum D-ASP in the DAA group, we showed significant increases in DDO levels in response to D-ASP supplementation. The degradative role of DDO is to catalyze the oxidative deamination of D-amino acids to generate the corresponding 2-oxo acids, along with hydrogen peroxide and ammonia (or methylamine).

In rodents, the administration of D-ASP was shown to increase DDO activity (Nagasaki. 1994; Yamada.1989), suggesting that DDO activity is induced by increased levels of D-ASP. Based on this information, in the present study, it is possible that because of the higher baseline levels of testosterone, as a means of androgen-regulated feedback of the HPG axis, the level of serum D-ASP induced by supplementation was conceivably being degraded by DDO at a rate that rendered it unable to effectively activate the HPG axis." (Willoughy. 2013)

Or to put is simply: If DAA works at all, it works only in guys like the participants of the 2009 study by Topo et al. who had baseline testosterone levels at the lower end of the normal range (4.5ng/ml) at the beginning of the study and average levels (6.4ng/ml) at the end of the study period. In trained athletes with testosterone levels in the range of 8ng/ml such as the guys in the Willoughby study the negative feedback will prevent any further increase in testosterone.

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Does testosterone actually build muscle and are natty test boosters worth your money? (learn more)

Bottom line: Before we have counter-evidence (this is science guys, you can't say something definitive based on one study) there is no good reason for someone with normal testosterone levels to spend money on D-Aspartic acid supplements.

Whether the same would be true for guys on post-cycle therapy or those who want to combat diet- / overtraining induced reductions in testosterone would certainly be worth investigating. Other aspects that could make a difference are the form of delivery (although this did not seem to be a problem; after all the DAA levels rose) and the provision of adjuvants to block the negative feedback on the HPG (see explanation above).

References:

• Nagasaki H. Gender-related differences of mouse liver-D-aspartate oxidase in the activity and response to administration of D-aspartate and peroxisome proliferators. Int J Biochem 1994;26:415–23.
• Shelmadine B, Cooke M, Buford T, Hudson G, Redd L, Leutholtz B, et al. Effects of 28 days of resistance exercise and consuming a commercially available pre-workout supplement, NO-shotgun, on body composition, muscle strength and mass, markers of satellite cell activation, and clinical safety markers in males. J Int Soc Sports Nutr
  2009;6:16.
• Spillane M, Schwarz N, Leddy S, Correa T, Minter M, Longoria V, et al. Effects of 28 days of resistance exercise while consuming commercially available pre- and post-workout supplements, NO-shotgun and NO-synthesize on body com position, muscle strength and mass, markers of protein synthesis, and clinical safety markers in males. Nutr Metab (Lond) 2011;8:78
• Topo E, Soricelli A, D'Aniello A, Ronsini S, D'Aniello G. The role and molecular mechanism of D-aspartic acid in the release and synthesis of LH and testosterone in humans and rats. Reprod Biol Endocrinol 2009;7:120
• Willoughby DS, Leutholtz B.  d-Aspartic acid supplementation combined with 28 days of heavy resistance training has no effect on body composition, muscle strength, and serum hormones associated with the hypothalamo-pituitary-gonadal axis in resistance-trained men.  Nutrition Research, Available online 15 August 2013. 
• Yamada R, Nagasaki H, Nagata Y, Wakabayashi Y, Iwashima A. Administration ofD-aspartate increasesD-aspartate oxidase activity in mouse liver. Biochim Biophys Acta 1989;990:325–8.