Disappointing Results in 28-Day Creatine + β-Alanine Study: No Performance Benefits, No Muscle Gain, No Fat Loss, No Increase in Phosphocreatine & Carnosine in 32 Women

Let's take a closer look at the study and find how it was possible that two proven ergogenics "failed".

Creatine and beta-alanine belong to the few "proven ergogenics", but according to the latest study from the University of Pittsburg, the Texas Christian University, the University of Wisconsin – La Crosse and the Texas A&M University they are not as effective as some of us may think. Specifically the effects of beta-alanine which was tested in what you may call its "comfort zone", i.e. a graded exercise test on the cycle ergometer for VO2peak with lactate threshold determination, and multiple Wingate anaerobic capacity tests. And still, the overall results of the study is that there a "no consistent additive benefits of BA [beta alanine] and CRE [creatine] supplementation in recreationally active women.

If you are using creatine already try adding bicarbonate as extra-cellular pH-buffer

The Hazards of Acidosis

Build Bigger Legs W/ Bicarbonate

HIIT it Hard W/ NaCHO3

Creatine + BA = Perfect Match

Bicarb Buffers Creatine

Beta Alanine Fails to HIIT Back

In today's SuppVersity article, we are going to have a closer look at the study design, its outcomes and potential explanations for the absence of the highly desirable performance enhancing effects of these two (alleged) ergogenic powerhouses.

As you may know I am not a fan of beta alanine, anyway. Yet despite my alleged bias, I have to admit that the wingate tests the scientists used to determine the effects of the supplementation protocol may have been too short for BA to work. In the most comprehensive meta-analysis of the research to-date, Hobson et al. (2012) found that there are no ergogenic effects to beta alanine on exercises lasting less than 60s or more than 240s; and in the "ergogenic" 60-240s zone, the performance benefit is only 2.85%.

Figure 1: In view of the short study duration it's no wonder that there were no significant effects on body fat and lean mass, but the fact that the beta alanine only group actually gained fat after an initial high loss of body fat is still awkward - still, statistically significant was only the time effect, which tells you that exercise works (Kresta. 2014).

And as far as the absence of benefits of creatine are concerned. The results of the study are in line with previous experimental evidence like that presented by Green et al. who report in their 2001 article in the The Journal of Strength & Conditioning Research that...

"[...] short-term Cr supplementation does not enhance MP and PP during repeated upper-and lower-body Wingate tests when not accompanied by an increase in body weight." (Kresta. 2001)

Similarly, Hoffman et al. (2008) could not find perfomance benefits of short-duration beta alanine supplementation in college football players, what the scientists from the College of New Jersey did find, though was an increases training volume and reduces subjective feelings of fatigue in their highly trained subjects in response to the ingestion of 4.5g/day of beta alanine (Hoffman. 2008).

All in all, the results are thus less surprising than they appear to be...

... at least for those of you who don't believe in the unsustainable promises of the supplement industry, but rely on experimental evidence, only. For creatine, the scientists tested the wrong type of exercise. For beta alanine the exercise duration (60s) on the wingate tests was not long enough to show significant performance increases.

Figure 2: Non-significant (!) changes in carnosine (should increase with BA supplementation) and phosphocreatine (should increase with creatine supplementation) in the BA, BAC, CRE and placebo group (Kresta. 2014).

What the previous brief review of selected experimental evidence does not explain, though, are (a) neither the beta alanine, nor the creatine or combined supplementation lead to statistically significant increases in carnosine (via beta alanine) or phosphocreatine (via creatine), (b) the levels of phosphocreatine the high energy resource, that is believed to be responsible for most of the beneficial effects of creatine actually dropped after 2 weeks on maintenance dose of 0.1g/kg creatine, when it was administered after a 0.3g/kg creatine pre-load. These results stand in contrast to previous studies, like...

• Harris and colleagues (2001) who reported that β-ALA supplementation (3.2 g/day) resulted in a 42% increase in muscle carnosine levels after four weeks of supplementation not due to the fact that the carnosine levels didn't increase, but rather due to the fact that the scientists did not find statistically significant interactions among groups in muscle carnosine levels.
  
  As Kresta et al. (2014) point out, "the lack of statistical significance was apparently due to the large variability in muscle carnosine levels observed in response to β-ALA supplementation, assay variability, and/or inadequate sample size", so that "[m]ore research is needed to determine the effects of β-ALA supplementation on muscle carnosine levels in recreationally-active women" (Kresta. 2014).
• Greenhaff et al. (1994) or Harris et al. (1992) who found significant increases in phosphocreatine with similar preloading + maintenance creatine supplementation schemes as the one used in the study at hand, but yielded significantly higher and above all consistent increases in creatine of up to 40% . Results from the present study found non-significant increases in muscle PCr of up to 40%
  
  Again, Kresta et al. suspect that "the lack of significance may have simply been a result of the small sample size", but add that "it is also known that there is individual variability in response to creatine supplementation" (Kresta. 2014) - a fact that is imho unlikely to be a likely cause of the lack of effect in all subjects, though.

Overall it is thus difficult to determine the lack of consistence improvements in carnosine and phosphocreatine levels in the study at hand, it may yet, as Kresta et al. suggest also be possible...

A study by Everaert, et al. indicates that women have naturally lower carnosine levels (Evaerart. 2011 | see figure abvove). Previous studies, e.g. Tallon (2006), however, found no such difference which is interpreted by Harris et al. in their 2012 review as evidence that "that the apparent gender difference reported by Everaert et al. (2011) may have been simply due to a higher type I:II ratio in females in the voxel sampled." (Harris. 2012)

"[...]that sex may have played a role in response to creatine and/or β-ALA supplementation. In this regard, most studies on creatine and β-ALA supplementation have been conducted on males and there is some evidence that females may respond differently to creatine and/or β-ALA supplementation. For example, Fosberg and colleagues (Forsberg. 1991) reported that females had greater total creatine amounts relative to tissue weight; however, other studies show there is no difference between males and females (Forsberg. 1991; Stegen. 2014).

There are also some data suggesting that men may have greater muscle carnosine levels than women (Derave. 2002; Harris. 2012); however, a recent study showed sex did not have an effect on increasing carnosine levels with supplementation (Stegen. 2014). Additionally, Bex and coworkers (2014) reported that carnosine loading is more pronounced in trained versus untrained individuals" (Kresta. 2014).

It is thus possible, but imho again not very likely that the fact that the subjects in the study at hand were women and or their individual training status may have had and impact on the hardly existing response to creatine and/or β-ALA supplementation.

Creatine + bicarbonate appears to offer a superior synergism | learn why

In the end, it's yet not the increase in carnosine or phosphocreatine that's important for us. What we are looking for are performance increases, which were probably absent due to the selected tests, on which previous studies have already shown that creatine and beta alanine have failed before to produce significant performance increases (see previous elaborations on the non-existent effects of BA on 60s and >240s exercise and the issue with creatine and wingate tests), plus changes in body composition for which the four-week study period may simply have been too short.

Against that background I would like to point out that the study at hand does not indicate that either beta alanine or creatine are useful. What it does, thought, is to remind us of the fact that (a) you won't see results over night and (b) even beta alanine and creatine are exercise-specific ergogenics and won't boost your performance an each and every type of exercise to the same extent. Or what do you think are the implications? Comment on Facebook!

References:

• Bex, Tine, et al. "Muscle carnosine loading by beta-alanine supplementation is more pronounced in trained vs. untrained muscles." Journal of Applied Physiology 116.2 (2014): 204-209.
• Derave, Wim, et al. "Muscle carnosine metabolism and β-alanine supplementation in relation to exercise and training." Sports medicine 40.3 (2010): 247-263.
• Everaert, Inge, et al. "Vegetarianism, female gender and increasing age, but not CNDP1 genotype, are associated with reduced muscle carnosine levels in humans." Amino acids 40.4 (2011): 1221-1229.
• Green, J. Matt, et al. "The effects of creatine supplementation on repeated upper-and lower-body Wingate performance." The Journal of Strength & Conditioning Research 15.1 (2001): 36-41.
• Harris, Roger C., et al. "The absorption of orally supplied β-alanine and its effect on muscle carnosine synthesis in human vastus lateralis." Amino acids 30.3 (2006): 279-289. 
• Harris, R. C., et al. "Determinants of muscle carnosine content." Amino acids 43.1 (2012): 5-12.
• Hobson, Ruth M., et al. "Effects of β-alanine supplementation on exercise performance: a meta-analysis." Amino acids 43.1 (2012): 25-37.
• Hoffman, Jay R., et al. "Short-duration< i> β</i>-alanine supplementation increases training volume and reduces subjective feelings of fatigue in college football players." Nutrition Research 28.1 (2008): 31-35. 
• Kresta, Julie Y., et al. "Effects of 28 days of beta-alanine and creatine monohydrate supplementation on muscle carnosine, body composition and exercise performance in recreationally active females." Journal of the International Society of Sports Nutrition 9.Suppl 1 (2012): P17.
• Stegen, Sanne, et al. "The Beta-Alanine Dose for Maintaining Moderately Elevated Muscle Carnosine Levels." Medicine and science in sports and exercise (2014).
• Tallon, Mark J., et al. "Carnosine, taurine and enzyme activities of human skeletal muscle fibres from elderly subjects with osteoarthritis and young moderately active subjects." Biogerontology 8.2 (2007): 129-137.

To Fail or Not to Fail - 5x10 or 10x5? The Energetic Demand of Your Workouts Doesn't Depend on Workloads, Alone

Image 1: PCr, ATP, whatever as long as there was energy left, Arnold kept pumping iron (pun intended ;-)

"To fail or not to fail", this question is probably about as ancient as the hilarious idea to engage in physical activity that is not in one way or another directly related to one of our two most fundamental needs, survival and procreation. Researchers from the Physical Education Department at the Sport Sciences University of the Basque Country and the Department of Health Sciences at the University of Navarra in Spain have recently examined this question from a slightly different angle than most of the articles you have probably seen in and on the various muscle mags and bodybuilding related websites on the Internet. What Esteban M. Gorostiaga and his colleagues wanted to know was:

Are There Significant Differences in Energy Metabolism When you Train to Failure?

Or, put another way: Does it make a difference if you fail from a molecular energetic point of view or is the mere number of reps the most fundamental determinant of the changes in muscle adenine nucleotides, inosine 59-monophosphate (IMP), phosphocreatine (PCr), creatine (Cr), lactate and energy charge during a workout. To answer this question the researchers recruited 6 healthy male volunteers (age 28-40y; BMI 23.3kg/m²; 1-RM_max on unilateral leg press 199+/-43kg) and had them perform a total of 50 repetitions with the same initial load (83% of 1-RM) on two separate occasions, either

• performed to failure, as a quintette of 5 x 10 (sets x reps), or
• stopped before failure, in a 10 x 5 fashion.

On both occasions the subjects rested 2 minutes between the sets. Furthermore, Gorostiaga et al. tried to eliminate "confounding factors", by equating the values of several variables such as initial load and total number of repetitions between both exercise sessions and making sure that whenever a "subjects could not lift the initial load during the following sets due to fatigue" the load was decreased by 15kg until the respective subject was able to complete all 50 repetitions (Gorostiaga. 2012).

Figure 1: Peak power output profiles (average for n=6 subjects) for each exercise during the two experimental conditions: when exercise was 5 sets of 10 repetitions to failure (10REP; open circles), and when exercise was 10 sets of 5 repetitions not to failure (5REP; filled circles; adapted from Gorostiaga. 2012)

It is plain to see from the data in figure 1 that performing all sets to failure (open circles) lead to a significant reduction in total workload (the area under the peak powder curve):

During 5REP all the subjects were able to complete all the repetitions with the initially load assigned (154+/-31 Kg; 83+/-8% of 1RM). During 10REP, however, most of the participants were unable to complete all the repetitions with this starting load, due to failure. The load had to be reduced by 7.2+/-3.8% after 27+/-16 repetitions and was progressively reduced, reaching 85+/-12%(P,0.05) of the initial load at the last repetition. Average load during the 50 repetitions of 10REP was 6.1+/-6.3% lower (P<0.05) than during 5REP. (Gorostiaga. 2012)

If we examine the graph further there are a couple of other interesting things to observe, though:

• the 2nd-3d rep was the one with the maximal power - so much about the value / validity of 1-RM maximum strength tests, then ;-)
• the power progressively declined from the 3rd rep on (35-45%) and that with an astonishing dip after the 5th rep - maybe because subjects are used to do 5 reps, so that this could also be a psychological factor
• while not training to failure with 10 sets of five reps allows to maintain almost identical average peak power on all sets, training to failure with 5 sets of 10 reps resulted in a net reduction of 33% from 812Watts on the first to 569 Watts on the last set
• the average peak power per set was accordingly 28% lower, when the participants trained to failure

What's the "energy charge"? The energy charge was calculated as the quotient of (ATP + 1/2 ADP)/(ATP + ADP + AMP) and does thus quantify the ratio of usable to used energy in the muscle samples.

Yet while the average mean power output changes paralleled those of peak power output in both experimental conditions, the opposite was the case for the aforementioned muscle metabolites - muscle adenine nucleotides, inosine 59-monophosphate (IMP), phosphocreatine (PCr), creatine (Cr), lactate and energy charge - the scientists measured by high-performance liquid chromatography from the muscle biopsies they had taken from the right legs of the subjects on each occasion.

Is light training an option, at all?

Very much in accordance with the subjective experience of many trainees, the data in figure 2 appears to confirm that working out to failure does induce muscular exhaustion, which manifests in the form of physically quantifiable changes in muscle metabolites in the training to failure group

• almost depleted PCr stores in the failure group (85% fall, P<0.05), and 
• reduced ATP (-21%), energy charge (-4%), and  
• reduced adenine nucleotides pool (-20%; ATP + ADP + AMP), in the presence of
• increased IMP (+8600%) and lactate (+1400%) levels

Which stand in stark contrast to the mediocre decrease in of phosphocreatine, the almost unchanged muscle ATP, IMP, energy charge and adenine nucleotide pool and comparatively marginal elevations in blood lactate in the 10 x 5 non-failure group.

Figure 2: Changes in ATP, ADP, AMP, phosphocreatine (PCr) inosine monophosphate (IMP) and lactate from pre to post workout in the 5 x 10 (failure) and the 10 x 5 (no failure) session (data calculated based on Gorostiaga. 2012)

As far as the correlation between these markers and the actual power output during the workout are concerned, the scientist say that they observed a ...

Figure 3: Relationship between muscle lactate concentrations and average peak power (from Gorostiaga. 2012).

• significant linear negative correlation (R²=0.59) was observed between the average changes in peak power output observed during the last two repetitions (expressed in percent of the initial two repetition values) and the decreases in ATP levels (expressed in percent of initial value).
• significant curvilinear negative correlation between the average peak power output changes observed during the last two repetitions of the first and last sets (expressed in percent of the initial two repetition values) and the corresponding levels of muscle lactate. 

From the curvilinear nature of this relationship (see figure 3) Gorostiaga et al. conclude that "when muscle lactate levels do not exceed the upper limit of 10–15 mmol/kg wet muscle, power output changes little from maximum values". The exact opposite is yet the case, when the lactate values exceed this critical upper value and the power output begins to decrease sharply.

Image 2: While training like a sissy will at best produce suboptimal results, maxing out on every set of every workout will work for max. 2-3 weeks until you will not just lose the gains you may have made but end up weaker and with less muscle in the hospital, when an injury or total burn-out forced you to finally see reason. Going to failure on one the last set of a selected exercise for each body group may be a way smarter, safer and more productive way. Combine that with planned 3-RM tests to gauge your strength progress and you should see some nice gains and can keep track of your strength gains without risking burn-out or injury and trust me this has little to do with being a sissy!

Implications: At first sight you may certainly argue that the study does not provide much novel information. If you do however compare the main results to common wisdom about various strength training regimen, the total depletion of the phosphocreatine stores in the "higher" rep group and the increase in IMP levels, of which Gorostiaga et al. rightly argue that they reflect the failure of ATP resynthesis to match ATP hydrolysis rates and eventually feed into the uric acid cycle (as reflected by the 19% increase in the failure group) put an emphasis on the often underestimated energetic and metabolic demand of training to failure. In this context, the pronounced loss of purines from the muscle, as it has also been observed by Hellsten et al. subsequent to profoundly lowered ATP levels in the course of a one-legged HIIT protocol on a cycle ergometer (Hellsten 1999), and the subsequent extraction of urate from the blood by the muscle to restore intramuscular urate levels (remember: urate acts as a free-radical scavenger during intense exercise; cf. Hellsten. 1997) may well be an overlooked factor, when it comes to assessing exercise recovery.

Still, while the former would suggest that you better avoid training to failure altogether and simply hit the gym for a "light" 10 x 5 workout everyday, the minor reduction in PCr (-15% vs. -80%) and the non-existent rise in plasma lactate and urate (no stress = no adaptation?) do indicate that frequent light training session will probably not result in the desired, or at least suboptimal muscular adaptations, which are the physiological bases for the strength and size gains, you are looking for (read all about "The Physiology of Building Muscle"). Moreover, the almost unchanged ATP/ADP ratio (see figure 3), which is the gauge by which AMPK works (cf. "Zoning in on AMPK"), could be the reason why many of the "sissy workout" studies report that strength training would not have the same / any beneficial effect on glucose tolerance, lipid levels and all the other standard parameters of metabolic health scientists usually measure in those trials.

Bottom line: While it is almost certain that you will out-train your own recovery potential by going to failure on every set of every workout, the results of this study put an emphasis on the fundamental difference between physical workloads and their immediate physiological effects (just a reminder, the workload, i.e. weight lifted x reps was identical for both groups). What we are still lacking to derive concrete reliable workout tips from data like this, though, are clear-cut mechanistic or at least probabilistic relationships between the short term effects of and the long-term adaptation to different workout regimen and their respective energetic demands... ah, and by the way, this goes for the incredible popular measurements of post-workout protein synthesis, as well. Until now, no-one can say how much predictive value temporary increases in fractional muscle protein synthesis actually have in terms of long(er) term muscle gains.

References:

• Gorostiaga EM, Navarro-Ame´zqueta I, Calbet JAL, Hellsten Y, Cusso R, et al. (2012) Energy Metabolism during Repeated Sets of Leg Press Exercise Leading to Failure or Not. PLoS ONE 7(7): e40621. 
• Hellsten Y, Tullson PC, Richter EA, Bangsbo J. Oxidation of urate in human skeletal muscle during exercise. Free Radic Biol Med. 1997;22(1-2):169-74.
• Hellsten Y, Sjodin B, Richter EA, Bangsbo J (1998) Urate uptake and lowered ATP levels in human muscle after high-intensity intermittent exercise.Am J Physiol 274: E600–E606.