3.2g of Beta Alanine Reduce Rate of Perceived Exertion, Increase Time to Exhaustion and Ventilatory Threshold. Vegetarians, Older People and Diabetics May Benefit Most.

Image 1: If you are into running, ladies, beta alanine is for you ;-)

Those of you who make sure that they are getting their highly educative daily dose of the SuppVersity *rofl* will be aware that today's blogpost is, once again, dealing with beta alanine. Contrary to yesterday's post, which dealt with its pharmacokinetics, we are today going to have another look at what kind of real world performance outcomes the average (female!) physical culturist can expect from taking at least 3.2g of the beta amino acid per day - a dosage that has been shown in previous studies to increase intra-muscular carnosine levels by 27–39% in fast- and slow-twitch muscle fibers, respectively (Baguet. 2009). And though I do not want to spoil things, I can already tell you that the results make it quite clear why beta alanine is not the next creatine.

Somehow ergogenic, yet not really antioxidant

For the study that was conducted at the Applied Physiology Laboratory at the University of North Carolina, study that was conducted by A.E. Smith recruited 24 "recreationally active" women, of which the authors state that they "engag[ed] in 3–7 days per week of aerobic, resistance or recreational activities, but were not highly trained competitive athletes". With a mean age of 21.8 years, a height of 165cm and a body weight of 61.5kg the subjects are thusly representative of the average young woman who goes to the gym to either get or keep in shape. I am specifically emphasizing this, because - at least in the early days - beta alanine was heavily marketed as "the creatine for women" who fear the water retention people still claim was an inevitable side effect of creatine supplementation.

Image 2: If you retain water, this is not due to creatine monohydrate. Either you are taking to much (creatine loading is a thing of the past) or you have bought a product with shitloads of carbs in it - in that case, chances are its not only water you are gaining ;-)

Does creatine supplementation inevitably lead to water retention and weight gain? Just because this myth is still perpetuated, especially among female figure competitors, I thought it may be worth addressing this again: Pure creatine monohydrate without the sugar and the other bullshit you will find in many creatine supplements does not necessarily lead to increases in either total body or water weight. A study by Rawson et al. showed only recently that the consumption of 0.03g/kg creatine for six weeks did not result in statistically significant changes in body weight or water in men or women, despite significantly increased plasma creatine concentration and enhanced resistance to fatigue during repeated bouts of high-intensity contractions (Rawson. 2011).

The women were advised to simply stick to their usual routine and to refrain from taking any supplements and medications except from their 2x800mg beta alanine tablets. The latter were to be taken 3x a day... so according to Cocker, they should have consumed 2x0.8g x3/day = 4.8g/day and not, as the scientists state "3.2 g daily". Now, according to Smith et al. this was the "required dosage" all participants met. I can however not say, whether this means that the third dose was optional... and this is not the only oddity in this study, where it is well worth to look beyond the assessments and conclusions of the authors.

At the beginning and the end of the 28-day supplementation period, the women had to perform a graded oxygen consumption test (VO2max) to evaluate VO2max, time to exhaustion, ventilatory threshold and establish peak velocity (PV), as well as a "non-damaging treadmill run (oxidative stress run) for 40 min at 70% PV [peak velocity]". Before, immediately after and in the 2-6h post running window total antioxidant capacity (TAC), superoxide dismutase (SOD), 8-isoprostane (8ISO) and reduced glutathione (GSH) were measured. In addition to that, heart rate and ratings of perceived exertion were recorded during the 40 min run. The two main metrics of the study were thusly the potential anti-oxidant effects (TAC, SOD, 8ISO, GSH) and the anticipated immediate ergogenic effects (VO2Max, time to exhaustion, heart rate and perceived exertion) of beta alanine supplementation.

Figure 1: Effect of 28 days of beta alanine supplementation on maximal oxygen consumption (VO2max), time to exhaustion during a graded exercise test (VO2TTE) and ventilatory threshold (VT) and qualitative practical significance (data and caption adapted from Smith. 2011)

If you now have a look at the my graphical rehash of the scientists own evaluation of the effect beta alanine supplementation had on VO2Max, the time to exhaustion (VO2TTE) and the ventilatory threshold (VT), you will have to concede that mean improvements of 0.28%, 6.6% and 3.7%, respectively, as well as the large discrepancies among the subjects (from beneficial over negligible to harmful) do not actually speak for beta alanine.

Figure 2: Effect of beta alanine supplementation on oxidative stress markers measured as total
antioxidant capacity (TAC) and glutathione (GSH) and the qualitative practical significance
for women (data and caption adapted from Smith. 2011)

Things get even more confusing when we take a look at the antioxidant effects of beta alanine. Not only were the levels of superoxide dismutase (SOD) and 8-isoprostane (8ISO) not different between groups, and the effect of beta alanine on the total antioxidant capacity (TOC) of the subjects negligible, the scientists' summary of the effects does even suggest that, after an initial amelioration of the negative effect of treadmill running on GSH, there was some sort of a "likely harmful" rebound 6h after the 40 min exercise bout. Before you do now flush your beta alanine stores down the toilette, I suggest you first take a look at the actual (absolute) effects beta alanine supplementation had on the exercise induced changes in GSH levels:

Figure 3: Absolute GSH levels (in µM) immediately before (pre), post, 2h and 4h after treadmill running in the placebo and beta alanine supplemented women before (pre) and after (post) the 28-day supplementation period (compiled based on data from Smith. 2011)

As you can see in figure 3, there was an (unexplained) increase in GSH in the course of the 28-day supplementation period in both groups. With 2%, the latter was statistically non-significantly greater in the beta alanine group, and the "likely harmful" effect of beta alanine supplementation 6h after the end of the treadmill-run is simply the result of a smaller increase in GSH, when you compare the pre- to post-supplementation levels at the 6h mark - and I guess, you would agree that a +27% increase in GSH is not exactly something that deserves to be called "likely harmful", wouldn't you?

All-clear: Beta alanine is not ergolytic ;-)

Now that we have gotten that straight, let's get to the last (and most) significant benefit the women in the beta alanine group had from taking the supplement: a statistically significant reduction in the rate of perceived exertion during treadmill running (cf. figure 4).

Figure 4: Rates of perceived exertion during 40 min treadmill running before (pre) and after (post) 28 days of supplementation with beta alanine or placebo; small graph: relative difference post supplementation in women receiving BA vs. placebo (data calculated based on Smith. 2011)

It goes without saying that being 18% less fatigued is something that could well be worth spending the roughly 7$ for a 28-day supply on (calculation based on a dose of 3.2g per day taken over 28 days and assuming you buy your beta alanine in bulk at one of the major suppliers). This, by the way, could be particularly true if you belong to one of the following groups, who have been found to have low intra-muscular carnosine levels, to begin with:

Image 3: Older people are only one of the three groups who are "at risk" of low carnosine levels and are thusly most likely to benefit from beta alanine supplementation.

1. vegetarians - a 2011 study by Evaraert et al. found that "Vegetarians have a lower carnosine content of 26% in gastrocnemius compared to omnivores" (Everaert. 2011); and according to another recent study, the soleus carnosine content of vegetarians was "non-significantly" reduced by -9% after 5 weeks of sprint training, while the same protocol elicited increases of +11% in omnivores (Baguet. 2011)
2. older people - Evaraert et al. found a linear decline (ca. -10% in 20 years) in carnosine levels with age (correlation r=-0.26; Everaert. 2011); and Stout et al. report a highly significant +29% increase in physical working capacity at the fatigue threshold in twenty-six men (n = 9) and women (n = 17) (age ± SD = 72.8 ± 11.1 yrs) who  had been supplementing with 800 mg three times per day for 90 days (Stout. 2008)
3. type-2 diabetics - according to Gualano et al. type-2 diabetics have "significantly lower carnosine content (−45%) in gastrocnemius muscle", a relative deficiency of which the scientists argue that it "may be partially associated with defective mechanisms against oxidative, glycative and carbonyl stress in muscle." (Gualano. 2011)

After all, it does yet not really matter whether you are a type-2 diabetic, a vegetarian or simply getting older, compared to many (if not most) of the other overpriced ergogenics that are advertised all over the web, beta alanine is certainly not only one of the cheapest, but also one of the most promising candidates for the 3rd place on your list of staples, where (whey) protein and creatine should nevertheless still occupy position 1 and 2, respectively. And the fact that it did not prove to be a potent antioxidant in this study need not really be a disadvantage, after all, we still do not know whether the exercise-induced oxidative "damage" is not what actually triggers the highly desirable adaptive responses (cf. previous posts on "hormesis"), we are all looking for, when we are hitting the gym.

Intra-Workout Supplementation: Increased Carbohydrate Oxidation with L-Arginine, Lower Fat Oxidation with Glucose & Lowest Rate of Perceived Exertion with Plain Water

Image 1: This bird certainly knows about the importance of adequate hydration ;-)

Have you been at the gym today? If so, what kind of beverage have you been sipping in the rest-periods between your sets, your sprints or during your regenerative (not fat burning ;-) "classic" cardio exercise? Was it Funky XYZ the latest and greatest intra-workout product on the market? If so, you better check out its ingredients, who knows maybe the "latest and greatest" turns out to be quite counterproductive towards the goals you have been setting after reading one of the last two installments of the Intermittent Thoughts? Let's assume you are the "Peter Griffin"-type of chubby - in that case, I hope that your Funky XYZ did not contain glucose, maltodextrin, waxy maize, or any other of the sugars of which the supp companies are going to tell you that they "superior" to the white poison your granny uses in her delicious muffins. Why? Well, according to a soon to be published study by scientists from the Massey University in Wellington, New Zealand, as little as 12g of glucose will reduce the amount of endogenous fatty acid (i.e. the stuff your body is using to hide your abs ;-) oxidation by -22%! Sounds terrible, doesn't it? Well, let's look at some details to decide whether those -22% will really make a difference and what effects the presence of l-arginine and l-glutamine in your intra-workout supplement could have had.

150 min @ 177 Watt + Glucose + (Glutamine or L-Arginine) = ???

Figure 1: Composition of the intra-workout supplement; sodium citrate base + 12g glucose (glucose) and additional 1g l-glutamine (Glu + L-Glutamine) or 0.1g l-arginine (Glu + L-arginine)

It stands out of question that adequate hydration is of utmost importance, when it comes to maximizing athletic performance (incidentally, the same is true, when it comes to "burning fat"). What athletes should drink before (pre-hydration), during (hydration) and after your workouts (re-hydration) is thusly one of the classic topics of exercise science and the recent study by D.S. Rowlands et al. is thusly probably #1001 on the never-ending list of investigations into the optimal mineral and nutrient composition of intra-workout drinks. For us, it is of interest, because it is one of the few which investigated the differential effect of the amino acids l-arginine and l-glutamine on substrate utilization, plasma glucose, lactate and sodium levels and rates of perceived exhaustion in eight male cyclists and triathletes during 150min (!) of cycling at 50% of the individually predetermined peak power (this is noteworthy, because 50% of their peak power equalled 177 W, which is not exactly "light" exercise), in the course of which the athletes consume 150ml of a fluid containing a 0.95g sodium base and either 12g of glucose alone or a combination of glucose and either 1g of l-glutamine or 0.1g of l-arginine (cf. figure 1).

Figure 2: Oxygen consumption (L/min) and substrate utilization (g/min) in 8 trained cyclists / triathletes during 150 min of cycling at 177W with 150ml of four different intra-workout drinks (data adapted from Rowlands. 2011)

As a seasoned student of the SuppVersity, it should not surprise you that the exogenous (i.e. from the outside) supply of glucose produced a -22% shift in substrate oxidation from fatty acids to the now more readily available carbohydrates (cf. figure 2). What you have probably not expected, though, is that the addition of the minuscule amount of l-arginine (which is btw. about what you will get with many of the proprietary blends in the still incredibly popular "NO-boosters") would promote this shift by increasing the total amount of oxidized carbohydrates by another ~10% over the 12g glucose solution alone.

Figure 3: Comparison of total / relative substrate utilization for the 12g glucose + 0.1g arginine, the 12g glucose and the water + sodium citrate groups (data adapted from Rowlands. 2011)

Now you are stunned, hah? So after all it is yet not your fault that you cannot see your abs. It's your NO-suppement! Well, not exactly. I mean take a look at the way I arranged the data in figure 3. You will probably acknowledge that the 12g glucose + 0.1g l-arginine group "burned" more energy - if you want it in calories (remember this is stupid ;-) 0.68kcal/min or 102kcal during the whole session and then come back to the -22% reduced fatty acid oxidation and lament: "But Dr. Andro, they burned 22% less fat than the water-only group! Now I know why I don't get lean." If that is your train of thought, I would invite you to continue the idiotic kcal number crunching and calculate on how much fat the poor l-arginine group would have missed to burn... well, it's the "exorbitant" amount of 170mg/min or - for the whole session 25.5g! While this may be more than one tablespoon of coconut oil, I guess you will probably admit that this probably is not the reason your abs are still covered by a thick layer of flabby adipose tissue, won't you?

Arginine reduces oxygen cost at the expense of glucose

Now, the real interesting findings of the studies are thusly not the changes in substrate utilization but rather the profound impact the addition of the two amino acids had on the lactate levels during the 150min of cycling (cf. figure 4) and the rates of perceived exertion (RPE).

Figure 4: Plasma lactate levels (mmol/L) in 8 trained cyclists / triathletes during 150 min of cycling at 177W with 150ml of four different intra-workout drinks (data adapted from Rowlands. 2011)

The latter (RPE), and this is actually quite surprising, were minimal in the water + sodium citrate group and maximal in the 12g glucose + 1g l-glutamine group (0.8 pts greater on a 0-7 scale). The RPE values of the arginine group, on the other hand, were only marginally elevated and that despite the significant increase in glucose clearance, which, by the way, has also been observed by McConell et al. (McConell. 2006) and Linden et al. (Linden. 2010). 

In view of recent studies such as Greer et al. (Greer. 2011), who observed a small, but statistically significant decreases in endurance during a strength training circuit in response to Arginine-Alpha-Keto-Glutarate (AAKG) supplementation, it is yet very unlikely that the observed effects of an arginine-enriched glucose containing intra-workout supplement observed in this study "have the potential to benefit endurance exercise performance" (which is what the scientists, much to my surprise, conclude). Another thing is yet more than likely, I would even say it is 100% certain: Neither the results of this nor of any future study will change the sales ranks on Bodybuilding.com & Co., where the purported NO-Boosters (and factual stimulants) still are the front-runners of the "TOP 10 selling products" ;-)

Tabata Workouts: Do They Work & How Energy-Demanding Are They? 14.5 Kcal/Min Sounds Nice, But You Must Earn It!

Tabata training is intense: So if you don't have the guts to do it on your own, find someone to suffer next to you. Trust me that'll keep you going, if you'd have long surrendered if you had trained alone. Some gyms even offer special courses.

Most of you will probably be familiar with the ultra-short + ultra-intense HIIT prescription that's known as the Tabata protocol. Not really? Well, here is the elevator pitch, then:  "Tabata training," was first described by the Japanese scientist Izumi Tabata in 1996. Tabata and his colleagues (Tabata.1996) conducted a study that compared moderate-intensity continuous training at 70% of maximal oxygen consumption (VO2max) for 60 minutes, with HIIT conducted at 170% of VO2max. The HIIT training consisted of eight, 20-second all-out exercise bouts followed by 10 seconds of rest for a total of 4 minutes of exercise. Based on what you have read about the contemporary HIIT research here at the SuppVersity, you will be aware that Tabata's protocol is more intense, but also much shorter than the currently favored HIIT regimen with their ~1-4min (sometime even 8min!) intervals at 80-100% intensity.

Now, the "original" study found that HIIT improved aerobic capacity to a similar degree as moderate intensity continuous training (aka LISS). Nevertheless, it did resulted in an impressive +28% increase in anaerobic capacity.

Tabata 2.0? Is it time for a modification?

Meanwhile Tabata training has evolved to include a variety of modes and exercises. What has always remained an essential characteristic of this type of HIIT training, though are the classic 20-10 patterns (i.e., 20 seconds of all-out effort followed by 10 seconds of rest).

According to Talisa Emberts, John Porcari , Scott Doberstein, Jeff Steffen and Carl Foster, the general physiological effects of this type of training are well documented. What would be lacking, however, is data on the the relative exercise intensity and energy expenditure of Tabata training (Emberts. 2013). Therefore, the purpose of the study at hand, which comes right from the labs of the Department of Exercise and Sport Science at the University of Wisconsin, was to determine the u exercise intensity and energy expenditure of a Tabata workout.

What did the scientists do?

In order to measure the energy expenditure, the researchers recruited 16 trained volunteers (8♂: 35.3 ± 8.1 years, 1.81 ± 0.06 m, 93.7 ± 8.70 kg, 53.2 ± 0.6 ml·kg·min -1; 8♀: 28.4 ± 9.3 years, 1.71 0.09 m, 71.9 ± 12.0 kg, 42.9 ± 11.3 ml·kg·min -1). After the usual initial fitness tests, each subject completed two identical workouts.

Table 1: .Exercises included in the 20-minute Tabata workout; each exercise was repeated twice at a ratio of 20 sec exercise/10 sec rest

• workouts consisted of four, 4-minute "segments".
• segments consisted of performing the exercises listed in table 1 twice in succession.  

• subjects completed as many repetitions of each exercise as possible in 20 seconds followed by 10 seconds of rest.

• there was 1 minute of rest between each segment.  

As Emberts et al. point out, they "chose to do the four segments of Tabata in succession, since one of the criticisms of Tabata training has been that individuals cannot burn a sufficient number of calories in 4 minutes to favourably impact energy balance" (Emberts. 2013)

Figure 1: Energy expenditure per minute and total energy expenditure in 20 vs. 4 min TABATA studies (Emberts. 2013; Olsen. 2013)

A brief glimpse at the data in figure 1 goes to show you that the relative energy expenditure per minute was as impressive as in previous studies:

"Caloric expenditure averaged 14.5 ± 2.7 kcal·min -1 , which is very similar to the value found by Olsen (2013), who reported a slightly lower value of 13.4 kcal·min -1 . This was probably due to the fact that her study included 13 women and only 3 men. Total energy expenditure ranged from 240 to 360 kcals for the 20-minute workout, which is significantly higher than the estimated 54 kcals expended during the 4 minutes of exercise reported by Olson." (Emberts. 2013)

What was also impressive, though, was the rate of perceived exertion (RPE), which averaged 15.4  ±  1.3 for the two workouts and was that rated as "hard" by these already trained subjects. I mean, ask yourself what an untrained individual would have been telling you after high knee runs, plank punches, jumping jacks, side skaters, rope jumping, in/out boats, line jumps, push-ups, burpees, russian twists, squats, lunges, mt. climbers, push-ups, split squats and box jumps... right probably nothing: The average sedentary inhabitant of the Western Obesity Belt would simply have collapsed after two exercises ;-)

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If you are a pro (and I mean "are" and not think of yourself as one) looking for an intense workout that will help you cut body fat and gain muscle at the same time, look no further. The cross-fit protocol Smith et al. used in a study I wrote about earlier this year has what you are looking for - unfortunately it has a similarly insane intensity: "From 16% to 8% Body Fat in 10 Weeks: Crossfit Workout Gets The Leanest Shredded - But Only the Fittest Survive" (learn more).

240-360kcal/s in 20 min - is that worth it? It stands out of question that trained individuals who are willing and able to give their muscles and more importantly their central nervous system the recovery time they will need after these workouts will greatly benefit from the intensity of a Tabata routine like this. This is after all, the "uncomfort zone" in which someone with years of training experience under his / her belt can still enforce adaptation.

Contrary to the trained athlete, for whom the total amount of energy expenditure is secondary to proving new "growth" (the word refers to general growth as in increases not just in muscle power or size, but also overall conditioning) stimuli, the rookie and even many intermediate trainees are however going to be overwhelmed by the physical (some also by the mental) demands of this workout. For him / or her, a combination of strength training and LISS (rookie, or someone trying to cut weight) or strength training and "regular" HIIT (advanced trainee) may be thus in fact be  better choice. Not necessarily because it would burn more calories, but rather in view of the fact that it is not as overtraining prone as a 20min session of Tabata training.

References:

• Olson M. Tabata  interval  exercise:   Energy  expenditure and post-exercise responses. Medicine & Science in Sports & Exercise 45. 2013; S420.
• Emberts T, Porcari J, Doberstein S, Steffen J, Foster C. Exercise Intensity and Energy Expenditure of a Tabata Workout. Journal of Sports Science and Medicine. 2013; 12:612-613

Up Your Coffee Intake & The Temperature of Your Baths to Get in Shape; Up Your Mood W/ Self-Paced Aerobics; Up Your Vitamin D in Time; Up Both Fat & Sugar For Diabesity

Ice cold baths are retarded. A cosy hot tub and a cup of coffee make a way better peak conditioning strategy.

+38% that's the SuppVersity Figure of the Week and it is the fat liberating advantage of the ingestion of 3mg/kg caffeine (in water) before sitting for 30 min in a hot bath (42°C; up to the navel) - yeah, you read me right:  The latest study from the Graduate School at the Department of Health Care of the Soonchunhyang University in the Republic of Korea is about the thermogenic effects of HOT (not cold) baths.

Even in the absence of additional caffeine ingestion the latter increases the amount of circulating free fatty acids by 52% (80% with caffeine) with the ~200-300mg of caffeine before the bath it almost doubles the amount of free fatty acids and increases the circulating leptin levels by 28% (vs. 5.6% w/out caffeine).

Whether the significantly more pronounced reduction in waist circumference (8.9mm vs 6.7mm) Tae-Wook Kim & Jeong-Beom Lee observed in their 9 male subjects (age ~26y; BF% 20%; no habitual caffeine consumers) is yet a result of real fat loss, remains highly questionable. However, even if that's not the case  sitting in a hot tub after having a large cup of coffee would still qualify as a potentially valuable peak-conditioning technique before a contest or photo shoot - or, for the less ambitious, before the high school reunion or the a first date ;-)

Aerobic workouts for a better mood

(Buscombe. 2013) You know these drill instructor classes, where the mostly female participants get their more or less over- and under-sized butts kicked ending up either severely overtrained or chit-chatting with the lady next to them? Yeah, I see - you know exactly what I am talking about ;-)

I guess Richard M. Buscombe and Helen Inskip from the University of London won't like my prelude to their study and there is in fact nothing bad about doing aerobic session like the ones in the Buscombe study, but both, the effects on body composition are about as intensity dependent as the affective changes, Buscome & Inskip measured in their study.

Figure 1: Psychological and physiological effect of 40 min or aerobics at different intensity (Buscombe. 2013)

As you can see in figure 1 the absolute affective change from pre- to post was clearly intensity dependent. Yet despite the fact that the intense workouts had the most significant (beneficial) effect on the feeling scales, the researchers are probably right, when they highlight that

"there was a positive pretest-to-posttest shift in the FS scores, irrespective of the intensity of exercise [and that this] finding supports existing literature advocating the role exercise plays in contributing to positive affective change" (Buscombe. 2013)

Still, in this group of 15 women (medium age 35.08) who think of themselves that they are "fit" or "highly fit", the high intensity (complete routine performed matched to a music with a 140bpm speed) has the greatest reward effect (pre- vs. post).

You have no idea, why I am mentioning the speed of the music here? Read up on the "Accustic Gear" post from 2011 (read more) and the corresponding research overview from April 2013 (learn more) to understand why the rhythm and tempo of the music matters.

In view of the significantly highly rate of perceived exertion and the frustration during the workouts, it does yet stand out of question that the "self-paced" routine, in the course of which

"[t]he participants were instructed that they could add or take away levels from their 'Step' aerobics box, as appropriate [and were allowed] alter the intensity of their session at any stage throughout the course" (Buscombe. 2013)

would be the preferable way to exercise on a regular basis - also, or rather especially because it does not entail the same risk of overtraining as the "push yourself to the limits" high intensity pattern.

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One thing to keep in mind, though, is the fact that not everyone is as willing and able to push him-/herself, when he or she is told that it was not necessary. For less ambitious individuals, the "self-paced" workout may thus well end up mirroring the low intensity routine, in the course of which the participants used only small body movements and minimized any lifting or propulsion actions, performing their workout in sync with a 125 bpm music.

Detrimental effects of vitamin D deficiency on the heart accumulate with time 

(Assalin. 2013) I know that I am already notorious for being the "anti vitamin D guy", but that is about as overgeneralized as the notion that everyone must be taking at least 4k of vitamin D per day. People who have been following the SuppVersity for some time now will be aware that I have always maintained that you go and check your vitamin D levels, supplement, if necessary and re-check after 3-4 months. After all, we all know how detrimental it is to be in a state of full-blown deficiency....

What we did not know - at least up to a couple of days ago, was how "fast" the detrimental effects of vitamin D deficiency can become potentially life-threatening. At least for rodents a recent study from the  São Paulo State University in Botucatu, Brazil, fills this important knowledge gap.

Figure 2: Effects of short (2 months; D2) and long (4 months; D4) on vitamin D deficient diet in the absence UV light on inflammatory markers, anti-oxidant status and heart morphology; data relative to levels after 2 months on the standard diet with 1,000IU vitamin D/kg chow (Assalin. 2013)

The scientists put a group of weanling Wistar rats under non-UV lights on diets containind either 1,000IU VD/kg of chow (C2 and C4) or a vitamin D deficient, but otherwise identical diet for two (D2) and four months (D4), respectively and observed:

• lower beta-hydroxyacyl coenzyme-A dehydrogenase activity and higher lactate dehydrogenase (LDH) activity, as well as 
• increased cytokines release, oxidative stress, apoptosis and fibrosis and 
• left ventricular (LV) hypertrophy and lower fractional shortening and ejection fraction 

in all vitamin D deficient animals. The differences in LDH activity, LV weight, right ventricle weight, and LV mass did yet not achieve statistical significance before the rodents had been kept on the vitamin D deficient diet for four months.

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Bottom line: Given the fact that the animals in the study did not consume any vitamin D and were - due to being exposed to evanescent (non-UV) light unable to produce even minimal amounts of vitamin D, the study results confirm that you can survive without supplements for a couple of weeks. On the other hand, they underline the importance of getting your vitamin D levels checked regularly. After all, we do as of yet have no idea to predict, where you as an individual are on the 25OHD spectrum ranging from deficient to over the tops - for some it may suffice to get regular sun exposure in the summer, for others 5,000IU of D + sunlight may still not suffice to keep their levels in the normal range: Don't be cheap! Get your levels checked.

What's the best diet to induce metabolic syndrome? The SAD diet!

(Pranpawit. 2013) The research design of the latest study from the Institute of Food, Nutrition and Human Health at the Massey University looks like Araya Pranprawit, Frances M. Wolber, Julian A. Heyes, Abdul L. Molan and Marlena C. Kruger wanted to participate in a competition, where those researchers win whose rodents gain the maximal amount of weight in the minimal amount of time. I mean, if you put the little critter on either

• control: 23% protein, 12% fat, 55% starch, 11% sugar
• high sugar (HS): 22% protein, 18% fat, 60% sugar
• high fat (HF): 16% protein, 60% fat, 20% starch, 4% sugar
• high fat + high sugar (HFHS): 19% pro, 40% fat, 41% sugar

the question that remains is not: "Are the rodents going to develop diabesity"?, but rather "Which group of rodents will die from the consequences of it first?" ... well, guess what, in the end this was more or less what the scientists wanted to find out.

Image  from NYC Dept. of Health & Mental Hygiene: The words on this poster from a 2009 campaign in the NY subway must be taken literally (learn why)!

Since published data with regards to diet‐induced metabolic syndrome in the SD rat model remain inconsistent, it is hypothesized that different types and amounts of diet as well as the period of time the animals are exposed to the experimental diet may significantly affect metabolic parameters in SD rats. Therefore, the objective of the present study was to compare the effects of consuming diets high in saturated fat, high in sucrose, and the combination of high saturated fat plus high sucrose, for a short time (4 weeks) and a long time (8 weeks), on selected markers related to metabolic syndrome in order to identify the optimal diet and experimental period for establishing metabolic disorder in the SD rat model (Pranpawit. 2013)

While the rodents in the HFHS and the HS diets consumed about the same amount of fat, those who had been randomized to the high fat (only; HF) diet obviously did not really enjoy their chow. At least weight-wise these rodents consumed significantly less food than their peers and still gained (due to the high energetic density of the chow) significantly more weight than the rodents in the "pure sugar" (HS) diet (see figure 3).

Figure 3: Weight and fat gain, as well as changes in Homa-IR (insulin resistance) and serum cholesterol; * indicates expressed in reference to control group at 2 weeks (Pranpawit. 2013)

As you can see the actual fat gain was slightly more pronounced in the high fat + high sugar diet (no this does not stand in contrast to yesterday's post on insulin not increasing fat storage, because we are not dealing with insulin sensitive critters here, but with severely obese pre-diabetics with elevated FFA levels, which do not require prior conversion to fat to be stashed away).

SAD, but true: "Human Study Shows: Three Days on "High Fat" Standard American Diet Produce Heart Healthier LDL Particle Profile Than NCEP  Approved Low Fat Diet" (read more).

"In all groups, fasting glucose, insulin concentrations (and insulin resistance (HOMA‐IR index) significantly increased between the 4 week and 8 week time points (p = 0.05; data not shown), indicating that these parameters increase with age regardless of diet. At 4 weeks, test diets had no effect on fasting glucose compared to CONT. HS, HF, and HSHF slightly elevated insulin and HOMA‐IR (Fig. 1). At 8 weeks, glucose was slightly elevated in test groups compared to CONT. HS had no measurable effect on insulin or HOMA‐IR, while HF slightly increased and HFHS strongly increased these parameters, although this did not reach statistical significance." (Pranpawit. 2013)

Somewhat surprisingly, none of the test groups differed from CONT as far as the weights of liver, kidney, pancreas, or caecum (data not shown) are concerned. The only, yet statistically non-significant difference was a "tendency for an increase in pancreas size with body weight" (Pranpawit. 2013). Moreover, rats on fat‐containing diets had markedly heavier abdominal white adipose tissues compared to CONT and this effect was unsurprisingly greater after 8 weeks on the diet compared to 4 weeks.

What's certainly interesting and an observation that flys right in the face of the generally held believe that it was the dietary fat that would precipitate to a messed up cholesterol metabolism is the fact that the rodents in the HS group showed elevated circulating cholesterol levels, as well. The often touted effect of sugar on the elevation of triglycerides, on the other hand, was absent unless the rodents received additional fat in their so that the combination of exogenous and endogenously produced (by fatty acid synthesis) fat reached a level that could not be compensated for by an increase in fat storage and/or oxidation.

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Suggested read: "Standard Am. Diet Has 'Optimal' Fatty Acid Ratio to Induce Diabesity. Plus: Doubling SFA Would Yield More Benefits Than Halving Them" (read more)

Bottom line: While we have to be careful about direct extrapolation of conclusions pertaining to the corresponding effect in human beings, it is still interesting to see that (a) the high sugar diet with literally no fat produces a "skinny, but sick" (remember those rodents were not fat) phenotype that is characterized by a messed up glucose and cholesterol metabolism in the absence of obesity and insulin resistance, that (b) taking fasting or postprandial glucose levels as a means to judge the insulin sensitivity is not generally accurate and overestimated the de facto non-existent insulin resistance in the high sugar group (HS), that (c) it's rather the rise in free fatty acids than the carbs that are responsible for both the developing insulin resistance (in the HF group in the presence of low fasting glucose; cf. (b)) and last, but most intriguingly that "the starch‐based control diet may also gradually cause impaired glucose tolerance" and is thus not exactly healthier for the glucose metabolism of the rodents than the high fat or even the high fat + high sugar diets.

If we had to pick a "winner" to be awarded the grand prize for the most metabolically damaging diet it would thus (without question) be the high fat + high sugar diet, which does - not much to your surprise as I would get - actually show a high resemblance to the SAD diet with a relatively high content of saturated and omega-6 fatty acids (from lard, in the case of the study at hand) and tons of simple sugar. The high fat (only) diet and the high sugar (only) diet, on the other hand lead to obesity and insulin resistance, respectively - both did thus fail to induce the key features of the metabolic syndrome, the presence both.

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That's it for today and aside from the obligatory information that you can find addition real short (and I mean "short" as in few words) short news on www.facebook.com/SuppVersity, e.g.

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  Most nuts deliver way less energy than the label or calorie tables tell you - for almonds, for example it's ~35%  less (learn more)

  More direct evidence for the free fatty acid - muscular glucose uptake connection: Reducing the amount of free fatty acids in the blood has immediate effects on the ability of the muscle (esp. of diabetics) to take up glucose (read more)
• The verdict on nuts: Not only are nuts not fattening, a recent meta review suggests that incorporating them into your diet may even yield reductions in BMI and more important waist circumference (read more)
• Stupid fools! "Reduced fat" labels are a signal to overeat... well, sort of at least the average customer is more likely to overeat, when the label tells him/her "this food is not as bad as the real deal" (read more)

there is just one thing left to do for me, which is to wish all of you a great Saturday morning, noon, afternoon and evening (whatever your clock may tell you it is now and will be in a few hours :-)

References:

• Assalin HB, Rafacho BP, Dos Santos PP, Ardisson LP, Roscani MG, Chiuso-Minicucci F, Barbisan LF, Fernandes AA, Azevedo PS, Minicucci MF, Zornoff LA, Rupp de Paiva SA. Impact of the Length of Vitamin D Deficiency on Cardiac Remodeling. Circ Heart Fail. 2013 May 24.
• Buscombe RM, Inskip H. Affective change as a function of exercise intensity in a group aerobics class. Journal of Exercise, Science & Fitness. 2013 [epub ahead of print]
• Pranprawit A, Wolber FM, Heyes JA, Molan AL, Kruger MC. Short-term and long-term effects of excessive consumption of saturated fats and/or sucrose on metabolic variables in Sprague Dawley rats: A pilot study. J Sci Food Agric. 2013 May 24.

The Latest on Glutathione Supplements - Yes, They Can Work; And Yes, They May Even be Beneficial for Athletes

Do you really need even more pills? The answer is "NO!" - even if GSH supplements actually seem to work.

If you'd asked me 2 years ago, I'd answered the question whether glutathione (GSH) supplements even work with a determined "I don't think so!" Meanwhile, there have been a handful of interesting papers which indicate that oral glutathione supplements could actually work.

The latest and one of the more interesting of these papers comes from the Graduate School of Life and Environmental Sciences at the Kyoto Prefectural University where Wataru Aoi et al. took the next step and tried to prove the significance of Kovacs-Nolan et al.'s (in press) and Park et al.'s (2014) finding that glutathione is intestinally absorbed and transported intact across the human intestinal epithelial wall in a rodent model and in humans.

The hormesis-concept says: Antioxidants are not always good for ya!

Is Vitamin E Good for the Sedentary Slob, Only?

NAC Impairs Anabolic Effects of Exercise

Vitamin C + E Hamper Gains in the Elderly

C+E Useless or Detrimental for Healthy People

Vitamin C and Glucose Management?

Antiox. & Health Benefits Don't Correlate

More specifically, the researchers hypothesized that glutathione supplementation may contribute to aerobic metabolism during exercise as a result of activated mitochondria via PGC-1α in skeletal muscle. The purpose of their latest experiment was thus to examine the effects of glutathione supplementation on muscle fatigue in exercise as a result of improved muscular aerobic metabolism.

After the scientists had randomly assigned a group of hairy subjects (=IRC mice) to one out of four groups: sedentary control, sedentary supplemented with glutathione (2.0%, 5 μL/g body weight), exercise control, and exercise supplemented with glutathione. The rodents who had been on the respective regimen for two weeks were subjected to a treadmill run at 25 m/min for 30 min. Immediately post-exercise, intermuscular pH was measured, and hind limb muscle and blood samples were collected to measure biochemical parameters. What the scientists found was:

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  Figure 1: The provision of glutathione was not without effect. More specifically it increased the mitochondrial builder PGC-1a and it's upstream protein AMPK (A,B), it reduced the decline in pH in response to exercise (C) and increased the mitochondrial DNA (D) levels (Aoi. 2015).

  Evidence of increased plasma fatty acid oxidation: Plasma NEFA after exercise was significantly lower with supplementation of glutathione compared with the control group (df = 31, F = 5.90, p < 0.01).
• Evidence of the H+ buffering effects of glutathione: The interstitial pH levels in muscle were significantly reduced by exercise (df = 31, F = 4.36, p < 0.001; Figure 1C). However, the pH following exercise of the glutathione group was significantly higher than that of the control group (p < 0.05).
• Evidence that glutatione supplements may improve mitochondrial health: PGC-1α was significantly higher with glutathione intake (df = 14, t = −1.88, p < 0.05). In addition, AMPK, an upstream protein of PGC-1α, was also significantly higher in the sedentary treated with glutathione group than in the sedentary control group (df = 13, t = −2.76, p < 0.05).

Now, rodent studies are nice, but human studies are the real deal, right? Well, luckily Aoi, et al. did an additional double-blind, cross-over study involving 8 healthy men (35.9 ± 2.0 y) who received either glutathione (1 g/d) or placebo for 2 weeks.

What is glutathione aka GSH & do I have to supplement? Glutathione (GSH) is an important antioxidant in plants, animals, fungi, and some bacteria and archaea, preventing damage to important cellular components caused by reactive oxygen species such as free radicals and peroxides. It is a tripeptide with a gamma peptide linkage between the carboxyl group of the glutamate side-chain and the amine group of cysteine (which is attached by normal peptide linkage to a glycine).

Now, the really important question is yet if you do have to supplement GSH straight. Studies like Lands, et al. (1999) indicate that cysteine rich protein sources can have similar effects. In fact, Lands et al. found significant increases peak power and 30-s work capacity, as well as increased lymphocyte GSH in response to supplementing with Immunocal, a high cysteine whey protein isolate for 3 months. Similar results have been observed with non-patented whey protein formulas by Shute et al. (2004) & Zavorsky et al. (2007). 

After the 14 day supplementation week, the subjects exercised on a cycle ergometer at 40% maximal heart rate for 60 min. Psychological state and blood biochemical parameters were examined after exercise and yielded interesting results:

• Evidence of decreased lactate production: There was a significant increase in blood lactate concentrations at 30 min after exercise compared with pre-exercise in the placebo trial (df = 41, F = 3.90, p < 0.05) but not in the glutathione trial.
• Maintenance of stable muscular glutathione levels: The free form of glutathione in plasma was not changed by either exercise or glutathione intake. In contrast, protein-bound plasma glutathione was significantly reduced by exercise in the placebo trial (p < 0.05) although the reduction was moderated in the glutathione-supplemented group.

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  Figure 2: It looks as if two weeks of 1g/day of GSH had potentially performance relevant effects on untrained individuals (Aoi. 2015).

  Reduced heart rates and reduced perceived exertion & inreased vigor: There was a trend for lower heart rates during exercise at 40 and 60 min in the glutathione trial compared with the placebo trial (Z = −1.47, p = 0.071 and Z = −1.26, p = 0.104, respectively). There was also a trend for a lower RPE at 50 min (Z = −1.44, p = 0.075) and a significant decrease at 60 min (Z = −1.78, p < 0.05) in the glutathione trial compared with the placebo trial. Furthermore, the Profile of Mood State vigor–activity factor after exercise was significantly higher following exercise in the glutathione trial compared with the placebo trial (Z = −2.11, p < 0.05) (Table 1). In contrast, the fatigue–inertia factor was significantly lower in the glutathione trial compared with the placebo trial (Z = −1.82, p < 0.05), while marked difference was not found in the VAS scores between trials (Z = −0.98, p = 0.163)

Overall, it does therefore appear as if the previously doubted oral glutathione supplements could - if they are taken chronically and in high amounts - in fact be useful.

Figure 3: In the Richie Jr. study the GSH supplements turned out to be powerful natural killer cell "anabolics", too (Richie Jr. 2015).

Bottom line: Yes, I may have erred when I said that glutathione supplements are a waste of money two or more years ago and I have no problem admitting it. The study at hand, as well as the results Richie Jr. et al. (2015) present in their latest paper which shows that the chronic ingestion of 250 or 1,000 mg/day will lead to significant increases of GSH levels in blood increased after 1 and 3 months and 30–35 % and 17-19% increased GSH levels in erythrocytes, plasma and lymphocytes in response to the high and low dose of GSH and 260 % increased GSH levels in buccal cells in the high-dose group after 6 months, it stands out of question that oral glutathione supplements work. Specifically in view of the fact that Richie Jr. et al. also observed significant reductions in oxidative stress and up to two-fold increases in natural killer cytotoxicity increased in their recently published GSH study.

All this does not change that glutathione is still not a must have supplement for all of us, though. What the results do, however, is to demonstrate that GSH supplements are not the waste of money previous studies which suggested that they were not even absorbed had suggested. In view of the fact that the subjects wer untrained and considering the role of GSH as "master-antioxidant", is is yet questionable, whether athletes and not chronically inflamed individuals are the ones who are going to benefit most from 1g of glutathione per day does yet appear to be questionable specifically in view of the fact that whey contains significant amounts of cysteine and has been shown to have glutathione boosting effects, as well (Bounous. 2000; Shute. 2004; Zavorsky, et al. 2007) | Comment on Facebook!

References:

• Aoi, Wataru, et al. "Glutathione supplementation suppresses muscle fatigue induced by prolonged exercise via improved aerobic metabolism." Journal of the International Society of Sports Nutrition 12.1 (2015): 7.
• Bounous, Gustavo. "Whey protein concentrate (WPC) and glutathione modulation in cancer treatment." Anticancer Research 20.6 (2000): 4785-4792.
• Kovacs-Nolan J, Rupa P, Matsui T, Tanaka M, Konishi T, et al. "In vitro and ex vivo uptake of GSH across the intestinal epithelium, and fate of oral GSH after in vivo supplementation." J Agric Food Chem. in press.
• Lands, L. C., V. L. Grey, and A. A. Smountas. "Effect of supplementation with a cysteine donor on muscular performance." Journal of Applied Physiology 87.4 (1999): 1381-1385.
• Park EY, Shimura N, Konishi T, Sauchi Y, Wada S, Aoi W, et al." Increase in the proteinbound form of glutathione in human blood after oral administration of glutathione." J Agric (2014):6183–9.
• Richie Jr, John P., et al. "Randomized controlled trial of oral glutathione supplementation on body stores of glutathione." European journal of nutrition (2014): 1-13.
• Shute, Max. "Effect of Whey Protein Isolate on Oxidative Stress, Exercise Performance, and Immunity." (2004).
• Zavorsky, Gerald S., et al. "An open-label dose-response study of lymphocyte glutathione levels in healthy men and women receiving pressurized whey protein isolate supplements." International journal of food sciences and nutrition 58.6 (2007): 429-436.