The Counterintuitive Catabolic & Pro-Diabetic Effects of Leucine Supplementation in Rodents on Corticosteroids

Not the mice from this study, but still a nice example of the effects of dexamethasone on skeletal muscle (right; Quin. 2012)

"Leucine-laced water + stress = insulin resistance" - This simple equation is the net result of a recent study by Nelo Eidy Zanchi and his colleagues from the Laboratory of Applied Nutrition and Metabolism at the School of Physical Education and Sports of the University of Sao Paulo in Brazil. Inspired by previous research which clearly indicated that leucine does not only have pro-anabolic, but also insulin sensitizing effects, Zanchi et al. speculated that the provision of adequate amounts of leucine would blunt the catabolic and pro-diabetic effects of 7 days of intraperitoneally injections of  dexamethasone, an artificial corticosteroid that's used to treat all sorts of inflammatory diseases.

Remember SuppVersity Rule of Smart Supplementation No. 2? Right. Specificity!

In order to test their hypothesis that leucine supplementation either in low doses in the drinking water or as higher dosed oral gavage would ameliorate the negative side effects of DEXA treatment, the scientists randomized groups of 10 male Wistar rats to six groups receiving either low dose or high dose leucine supplements with and without dexmethasone.

"During  the duration of the experiment, which lasted  seven  days,  DEXA (a synthetic glucocorticoid analogue that does not bind to plasma binding proteins) was given daily (at 9:00 a.m.) through intraperitoneal injection (5 mg/kg/day); control groups received  an equivalent volume of saline (0.9% NaCl). As DEXA was reported to decrease food intake, all groups were  fed the same amount of food (in terms of caloric intake) equal to the DEX group. Thus, differences among groups did not originate from different food intakes. We measured the caloric content of our standard chow (16.32 kJ/g) as well as leucine (25 kJ/g) in a calorimetric bomb (FTT Oxygen Bomb Calorimeter) in  order to avoid differences in the caloric ingestion between experimental groups and observed that the total caloric consumption was not statistically different among groups." (Zanchi. 2012)

The leucine was administered either in dosages of 0.068g/kg body weight per day (low dose) or 1.35 g/kg per day (high-dose) twice daily at 8:00  a.m. and  2:00  p.m. through gavage over seven days. And while the scientists had selected the high dose (LH) "to induce a maximal increase in muscle protein synthesis and insulin plasmatic levels", the dosage in the LL (=low leucine) group was too low to increase either muscle protein synthesis or plasma insulin levels. The third, non-supplemented control group received an NaCl (sodium) placebo, the volume of which was identical to the supplement to make sure that any possible volume-induced effects of oral gavage that could for example be induced by gastric expansion would not skew the study results.

"But leucine has been shown to be anabolic! So it must help."

Aside from the usual basal fasting glucose, insulin, tryacilglycerol (TAG) and HOMA-IR values, the scientists did also assess the motor performance of the animals by the means of two standardized strength and ambulation tests (Kennel. 1996; Anderson. 2004; Viera. 2008).

Figure 1: Effect of 7 days of low (LL) and high (LH) dose leucine supplementation with and with out dexamethasone on total body mass, soleus (slow twitch) and EDL (fast twitch) muscle mass in male Wistar rats (left; values expressed relative to non-supplemented control) and corresponding changes in mean ambulation and grip strength (right; Zanchi. 2012)

As you can see in figure 1 the supplemental leucine failed to reduce the negative side effects of dexamethasone. As far as the total body weight and the fast-twitch muscle mass (EDL) are concerned, you could even argue that the high dose treatment (DEX-LH) did even amplify the catabolic effects of the synthetic corticosteroid:

"Thus, leucine supplementation at both low and high doses did not counteract body weight loss in both food restricted (control groups) and DEXA-treated animals. Soleus muscle mass did not differ among groups. Leucine supplementation at  high doses  attenuated food  restriction-induced EDL muscle loss (CON-LH group) when compared with the CON-NS group  (p < 0.05). All DEXA-treated animals presented reduced EDL muscle mass when compared with the CON-NS group (p < 0.05), and leucine supplementation at both low and high doses of amino acid did not attenuate it." (Zanchi. 2012)

Now, you may well argue that the mere fact that the muscle weight was "statistically significant" reduced, this does not mean that these reductions would be physiologically significant and that the minimal differences between the DEX groups would not matter, anyway. If you just go by the data on the left side of figure 1, this is certainly right, if you do yet also consider the significant reductions in muscle function (figure 1, right) and the fact that all that happened within no more than 7 days, the overall result should actually remind you of the "Three Simple Rules of Smart Supplementation" - and here specifically the 2nd one: Specificity!

Figure 2: Time course of the dexamethasone-induced detoriations in fed serum glucose levels and ameliorative effect of low and high dose leucine supplementation (left) and effects of the treatment on fasting insulin levels and HOMA-IR (index of insulin resistance) at the end of the study (Zanchi. 2012)

In fact, the data in figure 2 only confirms the notion that things that you cannot define "good and bad", "black and white" and "beneficial or detrimental" without a context and the outcome you are expecting. If you are trying to keep the postprandial blood sugar in check, for example he addition of an effective (high dose) of leucine to the diet would appear to be a good idea. If, on the other hand, you are more concerned about insulin resistance, you would be better advised to use minimal amounts of leucine or simply refrain from supplementation altogether.

Figure 3: If the ingestion of bolus amounts of leucine is not helpful, lacing the water of the rodents DEXA treated rodents with leucine turned them into full-blown diabetics (Zanchi. 2012)

As these results clearly demonstrate the provision of additional leucine is not useful to counter the negative side-effects of synthetic corticosteroids. On the contrary, the negative effects on insulin resistance are apparently even augmented and the muscle function is further compromised by the purpotedly anabolic high dose leucine supplement.

And while the overall effects of the bolus administration may still be negligible, the scientists ingenious idea that the provision of similar amounts of leucine in the drinking water in a second follow-up experiment turned out to be "capable of inducing a massive diabetic state" (Zanchi. 2012; see figure 3 for the ensuing surge in fasting blood glucose levels) while decreasing the mass of the fast-twich EDL muscles even further.

Bottom line: Overall these results only confirm the simple, but often neglected truth that inductive reasoning is a futile undertaking in the realms of exercise and nutrition sciences: What is good for an athlete is rarely optimal for an obese person, the same diet that helps the obese lose weight, will make the athlete feel miserable, and lacing the drinking water of rodents on corticosteroids with the exact same amount of leucine that has had highly beneficial effects on the insulin sensitivity of diabetic rodents in previous studies (Guo. 2010) will not only fail to ameliorate the glucocorticoid-induced detoriations in blood glucose, it will even exasperate them.

So, does that mean you should not take your whey protein or BCAAs any longer? No, if you did that you would make the exact same mistake as someone who laces his water with leucine in order to avoid the catabolic effects of the synthetic corticosteroid he is taking for medical reasons. On the other hand, the results of the study at hand should make you re-evaluate the necessity and even benefits of guzzling BCAAs all-day long, at least if the reason for doing so is that you believe that you are so stressed that you would otherwise fall into a catabolic black hole.

That said, there may even be implications for the average pre-diabetic inhabitant of the Western hemisphere who is eating his hamburger and French fries on the parking lot of the local fast food restaurant, because he cannot make room to prepare and consume a real meal somewhere in his busy and stressful schedule. I mean, despite the fact that the aforementioned specificity principle does not allow for anything but a still to be verified hypothesis, it does at least appear not to far-fetched that this chronic endogenous stress, despite being very different from the "stress" that's induced by the administration of a synthetic corticosteroid that does not bind to serum proteins, could have similar negative modulatory effects on the purported benefits of chronic leucine supplementation ... but as I've said before, this would be something to investigate in another study. So unless you are actually taking dexamethasone for medical reasons, you are probably not at risk of developing diabetes due to a high amount of leucine in your diet.

In the unfortunate case that you are actually on synthetic corticosteroids, a previous study by the same group of scientists, in the same rodent model does suggests that three workouts with three sets of squats (10 reps each) per week may offer the protection against corticosteroid induced muscle loss decreased skeletal muscle GLUT-4 expression and insulin resistance, leucine does not have to offer.... well, at least as long as you abstain from leucine supplementation, because the latter had the exact same detrimental effects in the 2011 study where it was administered to one of the experimental groups in conjunction with resistance training as it had in these more recent experiments in the absence of any type of workout (Nicastro. 2011). 

References:

• Anderson,  K.D.; Abdul, M.; Steward, O. Quantitative assessment of deficits and recovery of
  forelimb motor function after cervical spinal cord injury in mice.  Exp. Neurol.  2004,  190,
  184–191.
• Kennel,  P.F.; Fonteneau, P.; Martin, E.;  Schmidt,  J.M.; Azzouz, M.; Borg, J.; Guenet,  J.L.;
  Schmalbruch, H.; Warter, J.M.; Poindron, P. Electromyographical and motor performance studies
  in the pmn mouse model of neurodegenerative disease. Neurobiol. Dis. 1996, 3, 137–147.
• Nicastro H, Zanchi NE, da Luz CR, de Moraes WM, Ramona P, de Siqueira Filho MA, Chaves DF, Medeiros A, Brum PC, Dardevet D, Lancha AH Jr. Effects of leucine supplementation and resistance exercise on dexamethasone-induced muscle atrophy and insulin resistance in rats. Nutrition. 2012 Apr;28(4):465-71. Epub 2011 Nov 12.
• Qin J, Du R, Yang YQ, Zhang HQ, Li Q, Liu L, Guan H, Hou J, An XR. Dexamethasone-induced skeletal muscle atrophy was associated with upregulation of myostatin promoter activity. Res Vet Sci. 2012 Aug 29.
• Vieira, N.M.; Bueno,  C.R., Jr.; Brandalise, V.; Moraes,  L.V.; Zucconi, E.; Secco, M.; Suzuki, M.F.; Camargo, M.M.; Bartolini, P.; Brum, P.C.; Vainzof, M.; Zatz, M. SJL dystrophic mice express a significant amount of human muscle proteins following systemic delivery of human adipose-derived stromal cells without immunosuppression.  Stem Cells  2008,  26, 2391–2398.  
• Zanchi NE, Guimarães-Ferreira L, de Siqueira-Filho MA, Felitti V, Nicastro H, Bueno C, Jr, Lira FS, Naimo MA, Campos-Ferraz P, Nunes MT, Seelaender M, de Oliveira Carvalho CR, Blachier F, Lancha AH, Jr. Dose and Latency Effects of Leucine Supplementation in Modulating Glucose Homeostasis: Opposite Effects in Healthy and Glucocorticoid-Induced Insulin-Resistance States. Nutrients. 2012; 4(12):1851-1867.

5-10% Weight Reduction From Set to Set For Hypertrophy, Heavy Leg Workouts for Cyclists, Garlic For 400% Higher Test/Cortisol Ratios & Max(!) 1g Vitamin C for Muscle Gains

7% increase in breast cancer risk for every 500g above "normal" birthweight for Scandinavian women. Weight is yet not all that counts, mommy's gestational diabetes and even a large body size also precipitate to later disease.

7% per 500g that's the increase in breast cancer risk, the female offspring of Scandinavian women will have, if they are born heavier than normal. This figure is the SuppVersity Figure of the Week and comes from a study I came across a couple of days ago (Troisi. 2012). The statistics are based on birth register data of women from Norway, Sweden or Denmark who were subsequently diagnosed with primary, invasive breast cancer (n=51419) and 10 controls for each case from the birth registries matched by country and year of birth (n = 514,190).

Contrary to what you may think, the birth weight does yet not pose as much of a risk to become obese later in life as being larger than "appropriate" for your gestational age does (Eyzaguirre. 2012). If you also consider that gestational diabetes has been linked with increased risk of metabolic syndrome in the offspring (Davis. 2012) and that obesity in itself is an independent risk factor for breast cancer (Patterson. 2012), these should be more than enough good arguments not to surrender to your occasional food cravings and laziness - pregnant or not.

It's not all in your genes, but most in your hands

Although some people would love, if this was the case, because they could blame their own misery on the mistakes other  may have made, our lives and health are not fully determined by our genes and/or the mistakes our mothers may or may not have made. As Poston and Foreyt wrote in 1999, already: "Obesity is an environmental issue." And we are lucky: It is in our hands to change the environment we are exposing ourselves to and thus influence which of our genetic disposals will become active and are  promoted and which of them won't. Now that's obviously not just the case for obesity, muscular hypertrophy would be another example. Irrespective of your genetic make-up your strength and muscle gains stand and fall with the way you train, eat and supplement... and guess what, all of these points will be addressed in today's installment of On Short Notice.

• Experimentally validated: 5-10% drop in weights per set is "optimal" for hypertrophy training (Medeiros. 2012) -- Scientists from the Laboratory of Physiology and Biokinetic at the Faculty of Biological Sciences and Health on the UNIG Campus V at Itaperuna in Brazil find: The average resistance trainee - in this case a young man aged 24.0±4.5 years with a body mass of 78.3±10.2 kg and a height of 177±7 cm - can remain in the hypertrophy range (10-12 reps to failure) for most of his sets, when he reduces the weight by 5-10% after each set.
  
  Whether this will also yield optimal gains was yet not within the scope of this 5-week study. What these results do however tell you is that you are not training hard enough if you perform all your sets with the exact same weight in the exact same rep range - well, unless you don't just like to listen to Super Human Radio, but are actually related to Superman himself ;-)

• 

  Sir Chris Hoy's legs are not as hilarious as those of the German Robert Forstemann (Robert is the right guy), but I am pretty certain their size and strength played a very important part in becoming the most successful Olympic track cyclist of all times (six gold and one silver Olympic Medal + 11 times world champion)

  Heavy leg training could make the difference between victory or defeat at the end of a cycling race (Hansen. 2012) -- In a soon-to-be-published paper, Ernst A. Hansen et al. report that the addition of 12-weeks of heavy resistance training in the form of 4 lower body exercises (3 × 4–10 repetition maximum) which had to be performed twice a week enhanced the cycling performance of highly trained cyclists by 7% compared to the training outcome of the subjects in a control group who simply followed their regular endurance-only, protocols:
  

  "Performance was determined as average power output in a 5-min all-out trial performed subsequent to 185 min of submaximal cycling. The performance enhancement, which has been reported previously, was here shown to be accompanied by improved pedaling efficacy during the all-out cycling. Thus, E+S shortened the phase where negative crank torque occurs by ~16°, corresponding to ~14%, which was more than in E (P = .002)" (Hansen. 2012)

  Since the test was conducted at the end of a 3h cycling session, it should be plain obvious that those 15% increases in torque will catapult the strength trained endurance athlete to the forefront on every final sprint.

• Human dose equivalent of ~0.1g/kg garlic per day could not just boost your testosterone and lower the high protein diet induced increases in cortisol, it could also improve the way your body utilizes dietary protein (Oi. 2012)-- Actually this is not a new study, but since Maxim was not happy with things "so yesterday" as the increases in HDL and LDL the Arabian scientists observed in the garlic study I have been talking about at the end of Thursday's SuppVersity Science Round-Up on SHR, I thought others may be as happy as Maxim will hopefully be to hear that there is more to garlic than "just" its beneficial effects on your heart.
  
  

  

  Figure 1: Higher testosterone levels, an amelioration of the high protein induced increase in corticosteroids and a 40% increase in net protein balance are unquestionably impressive results given the fact that the all those differences were brought about within 28 days and by no more than 0.1g/kg (HED) of "supplemental" garlic in form of heat dried powder that was added to the chow (Oi. 2001)

  In fact, I am almost sure that the >400% increase in the testosterone to cortisol ratio you will see if you take a closer look at the data in figure 1, is probably rather what Maxim would have liked to hear me talk about. Especially in view of the fact that this endocrine effects went hand in hand with a highly significant +60% increase in protein retention (figure 1, top right). Think about it, if only part of he protein that was now no longer excreted in the urine / feces would be used for protein synthesis this would entail exactly those hypertrophy effects you don't see with your average "scientifically proven" herb-based testosterone booster.
  
  Unfortunately, the scientists did only measure the body weight and visceral fat pads, not the actual muscle mass of the rodents,. But if you go by their ratios it is obvious that the high protein + garlic group were not just the heaviest, but also the leanest.
  
  With +11 % vs. +5% in both the medium and high protein diets, the animals on the low protein did yet exhibit the most profound benefits as far as the body weight / visceral fat ratio goes. Against the background that their net protein balance remained the same, this observation does actually suggest that the pro-anabolic effects of garlic are not solely a result of a decreased protein excretion (see figure 1).

  

  Table 1: Principal sulphur compounds of garlic preparations (Hammami. 2012)

  Warning: Don't live on garlic alone! While the provision of 0.8% garlic powder did have beneficial effects on testosterone production in the study at hand, there are a couple of studies which suggest that a diet with 15-30% of crude garlic (Hammami. 2008 & 2009), as well as the administration of Diallyl trisulphide in isolation (Qian. 1986) and raw garlic juice (e.g. 600mg/kg per day for 21 days in Fehri. 1991) can compromise testosterone production and/or testicular function. In view of the difference between 0.8% garlic powder in the diet of the rodents in study at hand and 15-30% of pure garlic in the diet of the animals in the Hamami studies, it is most likely that the effects were dose-depended, but in case you are interested in health benefits of specific sulfor compounds in garlic, the data in table 1 on the left may still come handy to pick "your" preferred form of garlic.
  Rather than that, it appears as if the human equivalent of 0.1g/kg body weight of heat dried garlic powder that contained a total amount of 5.05 mg/g of total diallylsulfide (0.05 mg of monosulfide, 1.0 mg of disulfide, 3.4 mg of trisulfide, 0.6 mg tetrasulfide) had the ability to improve the incorporation of dietary protein into muscles (and other organs).

 

• Study shows: Vitamin C supplementation does reduce skeletal muscle hypertrophy in response to chronic overload (Makanae. 2012) -- Despite the fact that it has not even been published yet, the paper by Yuhei Makanae et al. actually only confirms what more and more scientists have been speculating about within the last couple of years. The provision of high does of active antioxidants, and as it seems in particular vitamin C, blunts the hypertrophy response to skeletal muscle overload.
  
  

  

  Figure 2: 14-day of 500mg/kg  (HED 0.08g/kg) supplemental vitamin C blunt skeletal muscle hypertrophy in rodents (Makanae. 2012)

  As you can see in figure 2 the effect size was relatively small, but statistically highly significant (p < 0.01) and that despite the fact that the supplementation regimen (500mg/kg body weight; HED: 0.08g/kg body weight) was not even that much higher than what some "vitamin C enthusiasts" are taking on a daily basis in the futile (and useless) effort to boost their serum vitamin C levels to a concentrations your body does - probably not without reason - try to counter by increasing renal vitamin C clearance.
  
  As the data in figure 2 shows, the same homeostatic mechanism we know from humans worked in the rodents, as well - well, at least with respect to the serum levels. In the plantaris muscle of the supplemented group, on the other hand, there was a significantly higher accumulation of vitamin C than in the placebo group. This increase went hand in hand with an attenuation of the repressive effects the chronic overload of the muscle had on the expression of the catabolic protein atrogin-1 and the increases in the pro-anabolic protein Erk1/2 (p < 0.01) in the non-supplemented animals. Based on this observations and with reference to the results of previous studies and the fact that neither the water content of the muscle, nor a significant reduction in food intake in the vitamin C group could explain the observed differences, Makanae et al. conclude "that oral vitamin C administration attenuates plantaris muscle hypertrophy induced by chronic mechanical load." (Makanae. 2012).
  
  What the study does not answer, though, is the question whether the effects would be identical in a real-world training scenario, where the temporary, yet more intense wear and tear on the muscle could in fact be sufficient to induce skeletal muscle hypertrophy human despite vitamin C supplementation. But let's be honest in view of the fact that scientific evidence for ergogenic benefits of more than 1g of supplemental vitamin C  per day (in humans) is simply non-existent, the take away message from the study at hand should actually read: Do not escalate your vitamin C beyond the 1g per day, if you don't want to risk compromising the results of all the hard work you are investing into your training.

That's is, another installment of On Short Notice and the first day of the weekend approaching it's peak. If you still have some time before whatever your plan for Saturday night may be and feel like you could use some seconds on today's short news, I suggest you head over to the SuppVersity Facebook Wall and check out the latest news on

• 

  Ever thought about what green tea, grape seed, curcumin, cranberry, and tons of other Super Food have antimicrobial effects? Considering the LPS-influx from the gut turns out to be a major contributor to all sorts of diseases, I am curious about how much of their effects are actually mediated by the gut microbiome.

  The differential role of intramuscular lipids in trained athletes and sedentary slobs and how the difference between performance enhancement and insulin resistance it all comes back to getting your as off the coach (learn more)
• Metformin 2.0? Scientists have developed a hypolipidemic, anti-atherosclerotic, anti-obesity, and glucose lowering agent called ETC-1002 (learn more)
• Confirmed: Grape seed could be the go-to neuroprotector for diabetics - GSE administration was found to be able to ameliorate most of the biochemical altered parameters in diabetic rats (read more)
• Fermenting your own dairy? Just add some catechin rich teas and the lactobacilli will strive. Makes you wonder about the 'internal' probiotic effects of green and black teas, as well. Doesn't it? (learn more)

There will be more, don't worry - so feel free to check for updates either directly on the SuppVersity Facebook Wall or simply by taking a look at the navigation in the right under "SuppVersity Facebook Wall" from time to time. Obviously, you can also simply "like" the SuppVersity on facebook to make sure you don't miss anything.

References:

• Davis JN, Gunderson EP, Gyllenhammer LE, Goran MI. Impact of Gestational Diabetes Mellitus on Pubertal Changes in Adiposity and Metabolic Profiles in Latino Offspring. J Pediatr. 2012 Nov 10.
• Eyzaguirre F, Bancalari R, Román R, Silva R, Youlton R, Urquidi C, García H, Mericq V. Prevalence of components of the metabolic syndrome according to birthweight among overweight and obese children and adolescents. J Pediatr Endocrinol Metab. 2012;25(1-2):51-6. 
• Fehri B, Aiache JM, Korbi S, Monkni M, Ben Said M, Memmi A, Hizaoui B, Boukef K (1991) Toxic effects induced by the repeat administration of Allium sativum L. J Pharm Belg 46:363–374.
• Hammami I, Nahdi A, Mauduit C, Benahmed M, Amri M, Ben Amar A, Zekri S, El May A, El May MV. The inhibitory effects on adult male reproductive functions of crude garlic (Allium sativum) feeding. Asian J Androl. 2008; 10:593–601.
• Hammami I, Amara S, Benahmed M, El May MV, Mauduit C. Chronic crude garlic-feeding modified adult male rat testicular markers: mechanisms of action. Reprod Biol Endocrinol. 2009; 24:57–65.
• Hansen EA, Rønnestad BR, Vegge G, Raastad T. Cyclists Improve Pedalling Efficacy and Performance After Heavy Strength Training. Int J Sports Physiol Perform. 2011 Dec 2. 
• Hammami I, El May MV. Impact of garlic feeding (Allium sativum) on male fertility. Andrologia. 2012 Sep 3.
• Makanae Y, Kawada S, Sasaki K, Nakazato K, Ishii N. Vitamin C administration attenuates overload-induced skeletal muscle hypertrophy in rats. Acta Physiol (Oxf). 2012 Nov 26.
• Medeiros Jr HS, Mello RS, Amorim MZ, Koch AJ, Machado M. Planned Intensity Reduction to Maintain Repetitions Within Recommended Hypertrophy Range. Int J Sports Physiol Perform. 2012 Nov 19. 
• Oi Y, Imafuku M, Shishido C, Kominato Y, Nishimura S, Iwai K. Garlic supplementation increases testicular testosterone and decreases plasma corticosterone in rats fed a high protein diet. J Nutr. 2001 Aug;131(8):2150-6.
• Patterson RE, Rock CL, Kerr J, Natarajan L, Marshall SJ, Pakiz B, Cadmus-Bertram LA. Metabolism and Breast Cancer Risk: Frontiers in Research and Practice. J Acad Nutr Diet. 2012 Nov 2. doi:pii: S2212-2672(12)01426-8.
• Qian YX, Shen PJ, Xu RY, Liu GM, Yang HQ, Lu YS, Sun P, Zhang RW, Qi LM, Lu QH.  Spermicidal effect in vitro by the active principle of garlic. Contraception. 1986; 34:295–302.
• Troisi R, Grotmol T, Jacobsen J, Tretli S, Toft­Sørensen H, Gissler M, Kaaja R,Potischman N, Ekbom A, Hoover RN Stephansson O. Perinatal characteristics and breast cancer risk in daughters: a Scandinavian population­based study. Journal of Developmental Origins of Health and Disease, Available on CJO 2012.

The Acute & 24h Effects of 3 Types of High Intensity Circuit Training on Testosterone & Cortisol in Young Trained Men.

It's obviously to have the 24h effects on testosterone and cortisol than only those measured after the workout , but can we make solid conclusions based on the additional data?

In spite of the fact that the acute testosterone and cortisol response to exercise appears to have little direct effects on the overall training outcome (Schoenfeld. 2013), acute increase in cortisol and reductions in testosterone, i.e. a decrease in the testosterone:cortisol ratio is a classic feature of overtraining and can very well blunt, if not reverse the beneficial effects of exercise on your health and body composition.

Against that background a recent experiment that was conducted by researchers from the University of Chieti-Pescara in Italy could be of great interest to everyone who is performing high intensity interval training on a regular basis. Why?

Well, in contrast to previous studies, Blasio et al. investigated both the acute and 24h effects of a high intensity interval resistance training regimen in trained young men.

Learn more about building muscle and strength at www.suppversity.com

Tri- or Multi-Set Training for Body Recomp.?

Alternating Squat & Blood Pressure - Productive?

Pre-Exhaustion Exhausts Your Growth Potential

Exercise not Intensity Variation for Max. Gains

Battle the Rope to Get Ripped & Strong

Study Indicates Cut the Volume Make the Gains!

To characterize the effects on heart rate and hormonal responses the subjects, eight trained, healthy trained men (28.61 ±3.51 yrs), performed three different workouts which had the same exercises, the same load and number of repetitions for each exercise, but different exercise order, recovery and speed of execution.

• RANDOM workout: the assigned goal was to complete the assigned repetitions respecting only two duties. The first one was don’t stop until all of the repetitions were completed; the second was that there were no assigned order of execution of exercises and no assigned consecutive repetitions to complete.
  
  Participants were thus free to choose both the order of exercises and number of consecutive repetitions for each exercise (i.e. 2 repetitions of kettlebell swing, 10 repetitions of medicine ball slam, 20 repetitions of squat, 4 repetitions of spin with Bulgarian bag, etc.).
  
  No recovery period was assigned, except the time necessary to move from a station to another, and no speed of execution of exercises was assigned: participants were free to choose the preferred speed. 
• LADDER workout: respecting the following order of execution, kettlebell swing, medicine ball slam, spin with Bulgarian bag, squat, pull-up, burpee, participants had to complete the total repetitions according to a pyramidal scheme (e.g. 1st lap 10 repetitions at each exercise, 2nd lap 9 repetitions at each exercise) until the total number of repetitions of each exercise was executed.
  
  Each lap of the circuit was followed by 1 minute of recovery. No speed of execution of exercises was assigned: participants were free to choose the preferred speed. 
• AS SOON AS POSSIBLE (ASAP) workout : respecting the following order of execution, kettlebel swing, medicine ball slam, spin with Bulgarian bag, squat, pull-up, burpee, participants had to complete the total volume in six laps executed as soon as possible.
  
  During each lap participants had to complete the sixth part of total number of repetitions of each exercise without rest among exercises. Each lap of the circuit was followed by 1 minute of recovery.

Salivary samples were collected before and after each workout, at 11:00 p.m. and at 7:00 a.m. of the following day. Salive was also collected during a non-training day. Similarly, before and after the workout, plasma lactate was measured while a beat-to-beat heart rate recording was executed during each workout. Cortisol (C) and testosterone (T) were measured in salivary samples.

2h before the workouts the subjects who had to abstain from sexual intercourse, stimulants and alcohol from 2 days before to the experimental days and until 9:00 a.m. of the following day, consumed a standardized meal that was lower to 400 and consisted of 33 cl of water, 35 cl of orange juice and two 30 g energy bars (Power Sport Double Use, Enervit, Milan, Italy).

Let's look at the results

While the protocols elicited the same heart rate response (the major part of each workout was spent between 80 and 100% of maximal heart rate, confirming the high cardiovascular intensity of the workouts), they elicited different hormonal and lactate variations with the LADDER workout producing the lowest lactate increase and the RANDOM workout eliciting the highest lactate, cortisol and testosterone increases.

Figure 1: Relative changes in hormone and lactate concentration in response to the workouts (Di Blasio. 2014)

When C was considered in ratio with T no significant differences have been shown among workouts-induced variations. Results of the analysis of covariance, executed on significantly modified variables, confirmed that basal hormonal and lactate values did not influence their variations.

When they studied the effects of workouts on prolonged hormones production (i.e. until the morning following the morning, di Blasio et. al. found that observed that observed that

"C had both time (F=179.723; p < 0.001) and group × time effect (F=10.942; p < 0.001): while during non-training day there is a physiological decline of C production at 11:00 p.m., during training days its decline is not present but seems to have a continuous increase from 7:00 p.m. to 7:00 a.m." (Di Blasio. 2014)

For the testosterone production the authors found both time (F=443.340; p < 0.001) and group × time effect (F=3.254; p=0.008) even if the group × time effect seems determined by the samples collected at 7:00 p.m., so that the effects cannot be ascribed fully / exclusively to the workout.

Figure 2: 23h hormone profile after the RANDOM, LADDER, ASAP workouts on a control day (di Blasio. 2014)

What is most interesting, though, is the cortisol to testosterone ratio. It shows the greatest inter-group differences and could potentially be of great physiological relevance (Crowley. 1996). In that, the LADDER workout has the most negative effect, as it will totally blunt the natural decline of the C:T ratio at noon.

In case you're planning to incorporate circuit training into your schedule, make sure to have a huge chunk of beef after your workouts ;-) - "Post-Workout Steak "Supplementation" (135g of Lean Beef) Augments Improvements in Body Composition In Response to 8 Weeks of Circuit Resistance Training" | more

Bottom line: As usual, it is difficult to interpret the results in order to make concrete practical recommendations. The lactate and hormonal data does yet suggest that the "random" order, i.e. a training that involves a self-selected exercise order and rep speed, as well as little to no rest between exercises is the least, the ladder training, with its decreasing 10, 9, ... rep numbers and one minute rest between each lap of the curcuits is the most metabolically demanding workout.

Whether and to which extend this translates into an increased risk of overtraining, let alone increased muscle and strength gains, on the other hand, remains to be seen. In view of the overall effect on lactate levels and the C:T ratio, though, the study does suggest that you better be careful with high intensity circuit / interval resistance training sessions and give your body adequate time to rest and recover | Comment on Facebook!

References:

• Crowley, Michael A., and Kathleen S. Matt. "Hormonal regulation of skeletal muscle hypertrophy in rats: the testosterone to cortisol ratio." European journal of applied physiology and occupational physiology 73.1-2 (1996): 66-72. 
• Schoenfeld, Brad J. "Postexercise hypertrophic adaptations: a reexamination of the hormone hypothesis and its applicability to resistance training program design." The Journal of Strength & Conditioning Research 27.6 (2013): 1720-1730.

Science Round-Up Seconds: Vitamin E Succinate, How It's Extracted from Barley Leaves, Kills Cancer, Ramps up Growth Hormone & Spikes Prolactin. Plus: Testostosterone & Thyroid Hormone Decline Due To Plyometrics & HIIT

Regardless of all the hypocritical hoopla around his persona, Lance Armstrong has always been able to push himself like no one else. No wonder that intense plyometrics were part of his regimen.

If the SuppVersity Science Round Up was a meal, I guess you could say that Carl Lanore and I were sort of gluttonous, yesterday (click here to download the podcast, if you have not already done so). We almost raced from one topic to another and therefore all the good stuff from the list is gone already and I am a bit pressed on time to get some "private life" in, so that I am not psyched about the idea of writing about auxiliary stuff.

Against that background and in view of the fact that I felt that the pace of yesterday's show did not really leave enough room for some important details, I will stick to rehashing and expanding on the stories about Vitamin E succcinate and the detrimental effects of beating the crap out of yourself doing plyometrics or crazy HIIT workouts (too regularly), in today's installment of the SuppVersity Science Round-Up Seconds.

Let's see. Why don't we start at the end of yesterday's show?

• Vitamin E succinate the most potent anti-cancer tocopherol known to man. As you have heard on the show, vitamin E succinate attaches directly to a protein that's preferentially expressed in carcinogenic or pre-carcinogenic cells. It goes by the name α-Tocopherol-associated protein (TAP) and was found to be one of the major α-tocopherol binding proteins in serum, liver, brain and prostate. What has as of yet not been so clear, though, is that the expression of this protein increases with the malignancy of (breast) cancer (Tam. 2012). 
  
  

  

  Figure 1: Effects of alpha tocoperyl succinate alone (TOS), doxorubicin alone (DOX) or both (DOX + TOS) on cell viability in human MB231 breast cancer cells (my edits, original from Tam. 2012) - note: The effect was less pronounced in other cancer cells, so that it is reasonable to assume that the efficacy of the therapy will depend on the exact genotype of the cancer (for those tested in the study it was MB231 > SKBR3 > MCF 10A)

  When alpha tocopherol succinate binds to the protein on the cancer cells, this will either alone, or in combination with chemotherapy trigger apoptosis and cell death. It is as of yet not fully elucidated why vitamin E succinate is highly cancer-specific and leaves the healthy cells intact, but this could be related to the high metabolic rate and exuberant ROS production of cancer cells. There is however some research that would suggest that the cancer cells literally suffocate in their own radical oxygen specimen (ROS), which can no longer be cleared from the cell, due to the alpha-tocopheryl succinate induced displacement of ubiquinone from CII and the subsequent blockade of succinate dehydrogenase (SDH) activity (Dong. 2012).  If this hypothesis holds true it would therefore appear that long-term chronic supplementation with vitamin E succinate cannot be recommended until future studies on its general safety have been undertaken. As an adjuvant to chemotherapy, on the other hand, it could drastically reduce the dosage requirements during chemotherapy in specific types of cancer (see figure 1) and thus minimize side effects.
  
  You see, there is more to it than you can say in two minutes on the radio and this is why I will make sure we don't rush through the items that fast, in the next show. Ah,... of course the dietary source. I had almost forgotten about that one. As mentioned on the show, alpha tocopheryl succinate was originally extracted from Barley leaves. An while this may not be the first paper dealing with this "natural vitamin E analog", the one by Badamchian et al. is probably the one you will be most interested in.
  
  Published in the Journal of Nutritional Biochemistry the paper does not only describe the isolation of vitamin E succinate from green barley leaf extract (BLE)...
  

  "BLE [barley leaf extract] powder (50 mg/mL) was suspended in water and stirred for 1 hr at room temperature. The mixture was then centrifuged at 3000g for 30 minutes using a bench-top centrifuge. The pellet was discarded and the supernatant was pre-filtered through a Millipore DEPTH filter. The filtrate was then filtered through 0.45 I.tM mem- brane and stored at -20 ° C for HPLC or biological assays." (Badamchian. 1999)

  ... it does also shine another spotlight on its potential biological effects, as far as it's ability to increase growth hormone, but (unfortunately?) also prolactin in isolated anterior pituitary cells from female rodents:
  

  

  Figure 2: Prolactin and growth hormone release in anterior pituitary cells of female rodents after incubation with different amounts of green barley extract in which vitamin E succinate had been deterimed as the main ingredient before (based on Badamchian. 1999)

  It's really hard to estimate whether or not one of these effects would translate from a rodent cell in the petri dish to you or me popping a cap with vitamin E succinate everyday. That's particularly true in view of the fact that the underlying mechanism of the increase in GH and the imho more concerning increase in prolactin is neither mediated by increases in intracellular C-AMP, as it would be the case for GRF (old acronym for growth hormone releasing hormone), nor is it induced by the hydrolysis of polyhoshpoinositide, which is the underlying mechanism of the stimulative effect of TRH (thyrotropin releasing hormone). So basically we neither know how it works, nor do we know, whether the oral ingestion of vitamin E-succinate would be sufficient to produce serum concentrations in the pituitary that would be high enough concentrations to make any difference at all (note: the scientists excluded the influence of other components of the extract by testing alpha tocopherol succinate on its own in a separate trial)
  
  Bottom line: Based on roughly one dozen of in-vitro studies there is simply still to little evidence to decide who, outside of people with a history of cancer or someone who is just undergoing chemotherapy would benefit. Therefore, I suggest you wait before you add vitamin E succinate to your list of 'must have' supplements. Is it promising? Sure! Is it exciting, yeah! Is it save for a healthy being to be taken chronically??? I can't tell.
• The detrimental hormonal effects of pushing yourself beyond the tolerable threshold - Hardcore plyometrics and heavy HIIT and their impact on testosterone, cortisol, thyroid hormone and co: I guess you did already get the main message when you listened to the show, but just to give you an idea about the actual quantities, I thought it would be nice to provide you with two graphs as a reference.
  

  

  Figure 3: Comparison of the hormonal responses measured in the plyometrics (left) and the HIIT vs. LISS (right) study (based on Ozen. 2012 and Hackney. 2012)

  If you focus mainly on the differential cortisol responses in the two studies, it would appear likely that we are dealing with two very different forms of 'overtraining' here. While the HIIT protocol (90s at 100-110%, 90s active recovery at 40% matched for workload with steady state jogging at 60-65% of the VO2 max) probably wouldn't be a problem, if the athletes would get adequate rest and nutrition in the days after the session, the 6-weeks of plyometrics (15 session, increasing density, 90-195 reps per session) were enough to send the participants right into the vicious circle of the Athlete's Triad (if you have not done so already, I suggest you read up on that in the eponymous SuppVersity series).
  
  And you know what? Despite, or I should probably rather say due to their compromised hormone levels the guys in the plyometrics study did not lose a single gram of body weight. Good for their muscle, bad for the fat which was likewise preserved by the hormonal shut down, which affected both cortisol and testosterone in a similar way. So is that good or bad news? Well, let me say it this way:. Usually I see people training for a purpose and while the outcome often is stagnation and chronic fatigue, I would suspect that only few of you will have that on their mind, when they are hitting the gym, right?

Apropos viscous circle, and overtraining in order to avoid "overblogging" I will call it a day for today. Come back tomorrow for a couple of wholly new studies from the realms of exercise and nutrition sciences and in case you are planning to drink this evening, I highly suggest you check out the SuppVersity Facebook newspost on the effects of green tea extract on the uptake of alcohol. It may well be that those old fatburner caps of yours can be put to a way better use ;-)

References:

• Badamchian M, Spangelo BL, Bao Y et al. Isolation of a vitamin E analog from green barley leaf extract that stimulates the release of prolactin and growth hormone from rat anterior pituitary cells in vitro. Journal of Nutritional Biochemestry. 1994; 5: 145-150.
• Dong LF, Low P, Dyason JC, Wang XF, Prochazka L, Witting PK, Freeman R, Swettenham E, Valis K, Liu J, Zobalova R, Turanek J, Spitz DR, Domann FE, Scheffler IE, Ralph SJ, Neuzil J. Alpha-tocopheryl succinate induces apoptosis by targeting ubiquinone-binding sites in mitochondrial respiratory complex II. Oncogene. 2008 Jul 17;27(31):4324-35. Epub 2008 Mar 31.
• Hackney AC, Kallman A, Hosick KP, Rubin DA, Battaglini CL. Thyroid hormonal responses to intensive interval versus steady-state endurance exercise sessions. Hormones (Athens). 2012 Jan-Mar;11(1):54-60.
• Ozen, SV. Reproductive hormones and cortisol responses to plyometric training in males. Biol Sport.2012; 29 (3).
• Tam KW, Ho CT, Lee WJ, Tu SH, Huang CS, Chen CS, Lee CH, Wu CH, Ho YS. Alteration of α-tocopherol-associated protein (TAP) expression in human breast epithelial cells during breast cancer development. Food Chemistry. 2012 [ahead of print]

Reishi Protects Against Cancer & Contains Anti-Androgen; Adrenalin Rejuvenates Brown Fat; Mild Stress Normalizes Cortisol / DHEA Ratio in Elderly; DHEA, Aromatase Inhibitors & BPA vs. Joint & Brain Health; Vitamin E Battles Lymphoma

I am not happy with how the short news on Facebook simply disappear into oblivion.

The SuppVersity figure of the week is "920"! "920" as in "920 published posts" here at the SuppVersity. The reason that's the figure of the week is that I have been thinking about ways to reorganize the archive, but am a bit lost on how to structure things in a better way using blogger. This beast simply isn't made for anything that goes beyond a weekly classic blogpost a la "last week I did this and that, read about..." *yawn*

What's yet even more enervating is that despite having the huge advantage of being easily posted and directly accessible for all of you, the tons of short news items I post on a daily basis on the SuppVersity Facebook Wall simply disappear into the 'Facebook nirvana'. I currently cannot spend any time on those technical / organizational matters, but in the course of 2013 things are going to change.

In the mean time, follow the SuppVersity on Facebook and read the news, right when I post them + use google to find articles you are looking for. You will be laughing, but that's the way I dig through those 920 posts, as well ;-) Simply type whatever you look for and add a "+site:suppversity.blogspot.com" to it.

Chinese fungi are laden with cancer-protective molecules and 5a-reductase (DHT) inhibitors

In view of a recent paper from the Universitat de Valencia (Rios. 2012) it appears that it would actually be hard not to decrease your risk of developing cancer if you include extracts from Ganoderma lucidum and other related fungi, such as Poria cocos, Laetiporus sulphureus, Inonotus obliquus, Antrodia camphorata, Daedalea dickinsii, and Elfvingia applanata in your supplement regimen. With a total of 81 compounds from Ganoderma lucidum and other species from this genus, as well as 96 compounds isolated from other fungi, principally Poria cocos. It appears that these creatures (fungi are no plants, neither are they animals, but still sort of living beings) could hold the key to hundreds o natural anti-cancer agents.

Ganoderma lucidum better known as Rheishi mushroom may have cancer protective effects, but does not taste so bitter that no sane human being would eat it for a reason (photo by Eric Steinert)

I guess, I don't have to tell you that Chinese Medicine, of course, knew about the powerful anti-inflammatory and anti-proliferative effects of the lanostanes in these fungi, all along. Intuitively and probably by trial and error, the Chinese have found the tetracyclic triterpenoids that are derived from lanosterols in these fungi to be effective in the treatment of various diseases including different types of cancer. As of late the mostly anecdotal effects are backed by more and more research. A preliminary study by Sliva et al. (Sliva. 2002), for example, found that a hot water extract of both spores and the dried fruiting body of G. lucidum inhibited the progression of cancerous growth by reducing the expression of constitutively active growth and inflammation promoting transcription factors AP-1 and NF-κB: 

"The extracts also inhibited the secretion of uPA, thereby suppressing the migration of breast cancer MDA-MB-231 and prostate PC-3 cells. [...] High levels of both uPA and uPAR are associated with advanced tumors and decreased survival time in different malignant human cancers." (Rios. 2012)

These benefits do however come with a downside. At least in vitro, some of the fungi, particularly G. lucidum, obviously exert their effects via anti-androgenic pathways (Liu. 2007). A compound in Reishi that goes by the name ganoderol B, for example, does not just have inhibitory activity against 5α-reductase, but can also bind to the androgen receptor directly (if you will you could say it acts like an anti-anabolic SARM). Now, this was good news in a study that has been conducted by Liu et al., because it inhibited androgen-induced cell growth in an LNCaP cell line while suppressing testosterone-induced regrowth of the ventral prostate in rats. If those effects are however non-selective, you are about to run into problems once the molecules bind to receptors outside the prostate -- epression for example, if they block the androgen effects in the brain, etc.

That reminds me: What did Carl say on Thursday's installment of the SuppVersity Science Round Up? "The good thing about supplements is that they work, the bad thing about them is that they work!" I guess you can basically say the same about Reishi. Good that you as a SuppVersity reader always know about both, the good and the bad sides of supplements!

More short news

With me posting stuff on the fly on facebook, I am actally 'wasting' many of the very short news items, but nevertheless, here are a couple of relatively short news to round things up.

• 

  If you missed Monday's new on the quasi non-existent thermogenic effects of ephedrine, this would be a good time to read that post.

  Stress-induced browning of the fat? Chronic overexpression of noradrenaline (re-)generates wasted brown fat! In a way this is a follow up on the ephedrine post "Fat Burners Don't Work" as well as an addendum to a news on the role of GABA and brown fat in obesity, I just posted on facebook. According to an allegedly old study in the International Jornal of Obesity (Lee. 1986),  the chronic overexpression of noradrenaline as it is observed in humans with a certain form of adrenal tumor known as phaeochromocytoma can actually reactivate the intra-abdominal fat of human adults, including the omental fat, which is brown adipose tissue in infancy.
  
  Lee et al. see this as one of the main contributing factors to the weight loss which is typically seen with phaeochromocytoma. Sounds, logical, since the same stimulus that rejuvenates the Brown fat will also have it burn energy continuously - the overexpression of noradrenaline.
  
  
  Symptoms of overtly high noradrenaline levels include abdominal pain, chest pain, irritability, nervousness, pallor, palpitations, rapid heart rate, severe headache, sweating, hand tremor, high blood pressure, sleeping difficulties, and also weight loss.
  Even patients with Cushing's syndrome (hypercortisolemia) there is an increase in brown fat compared to healthy individuals. That the latter is not as profound is probably due to the ameliorative effect of cortisol on nor-adrenaline. After all, cortisol comes into play, when the stress becomes chronic and the acute nor-adrenaline response to stress, when it was prlonged any further would actually pose a direct threat to your health (just as phaeochromocytoma does, by the way; see red box on the right)
• Repeated moderate stress exposure increases DHEA production and normalizes corticosteriod levels in old apes (Goncharova. 2012). Yep, you are reading right we are not just talking about no rodent studies (although macacs are not exactly very human either ;-), but also about the role of repeated moderate stress in the normalization of aged induced abnormalities in the expression of adrenal hormones.
  

  

  Figure 2: Cortisol/DHEA ratio before and after 2h/day of immobilization stress.

  "In old monkeys the basal DHEAS levels were lower, while the [cortisol]/DHEAS ratio was higher than in young animals. Repeated immobilizations inhibited [cortisol] elevation on day 3, caused no changes in DHEAS reaction, led to increase of basal DHEAS levels and to a reduction of [cortisol]/DHEAS ratio on days 2, 3, 4, 10, 11." (Goncharova. 2012)

  In figure 2 you can see how profound the differences between acute, subchronic (3-day) and chronic responses actually are and that after 10 days of daily stress exposure in the form of daily 2-h immobilization stress actually are. Since I assume you don't want to be bound or enchained (well, maybe you want?), I suppose a viable alternative could be a shorter not too intense workout, although the acute responses to the latter vary with age as well (cf. Lennartsson. 2007).
• Monitor your DHEA levels closely, if you are concerned about joint degeneration According to a study that's about to be published in the next issue of the Journal of Steroid Biochemistry and Molecular Biology, DHEA, or the estradiol your body generates from it via local aromatization, exerts major protective effects against osteoarthritis (Li. 2012).
  
  
  An additional note of caution with respect to the abuse of aromatase inhibitors, natural or not. Since they have the potential to reduce the expression of estrogen at the neuronal level in the brain they can precipitate  Alzheimer's dementia. While estrogen appears to decrease the MMP-3 & 13 expression in the cartilage reasearch by Merlo et al. suggests that it will increase others, namely namely MMP-2 and MMP-9  in the brain and thus facilitate the clearance of the ameloid beta plague that's rendering the brains of AD patients more and more dysfunctional (Merlo. 2012). You see, it's no chance that pre-menopausal women are protected from Alzheimer's yet more susceptible to multiple sclerosis (MS). After all, high MMPs 7 & 9 have only been observed in kids with MS, as well (Unsal. 2012).
  In their experiments on a rabbit model of osteoarthritis the scientists from the Zheejiang University in China tried to nail down the beneficial effects of DHEA on chrondocites and cartilage to estrogen by co-administering DHEA with the aromatase inhibitor letrozole, and/or the estrogen receptor inhibitor fulvestrant and observed that the
  

  "[e]xpression of MMP-3 and MMP-13 increased in both DHEA-treated chondrocytes and cartilage in the presence of letrozole and/or fulvestrant, while the expression of TIMP-1 and collagen type II (Col-II) decreased." (Li. 2012)

  With the former metalloproteinases (MMPs) being proteolytic enzymes, which break down cartilage and the latter, i.e. the tissue inhibitors of metalloproteinases (TIMPs), acting as their antagonists, it appears clear why both patients on testosterone replacement therapy who use too high of a dose of aromatase inhibitors and athletes who abuse respective products on cycle or during PCT often suffer from severe cartilage degeneration - I mean, combine the endogenous cartilage destruction due to high MMPs and low TIMP levels with the wear and tear of weight lifting... what good could come out of that?
  
  Note: A very similar effect has been reported for bisphenol A by Wang et al. in 2010, already (Wang. 2010). No wonder, after all BPA decreases the local expression of aromatization in joints and cartilage so that less estrogen will be floating around to keep the MMP levels in check and TIMP up (Watanabe. 2012).
• Alpha tocopherol to prevent lymphoma Regular vitamin E, i.e. the alpha-version of the tocopherols is no longer the star at the supplement sky it has once been hailed to be. A recent study by Renu Sharma Manjula Vinaya that's been published ahead of print in the journal Molecular Biology Reports shows however that this does not mean that it's outdated and useless (ask Ray Peat about it ;-).
  
  

  

  Figure 3: Lifespan (top) and ascite volume (=water accumulation in the abdominal area) of lymphoma carrying mice treated with what would be in human terms ~325, 650 and 975 IU/day of an alpha tocopherol only supplement  (Sharma. 2012)

  In order to test the hypothesis that the ROS scavenging abilities of vitamin E should help with cancer prevention (as a student of the SuppVersity you know that respective data from epidemiological studies are equivocal, some suggesting the exact opposite; read more), the researchers initially induced the growth of lymphomas in 10-15 week old male mice and subsequently treated them with either 1.5 mg (50 mg/kg bw), 3 mg (100 mg/kg bw) or 4.5 mg (150 mg/kg bw) of alpha-tocopherol for 14 days.
  
  As the data in figure 1 goes to show you, the treatment increased the lifespan of the rodents by 25% and reduced the ascite fluid volme (see image in figure 1) by ~46%. This was accompanied by reductions in protein carbonylation and increases in the "master anti-oxidant" GSH, as well as several other markers showing that vitamin E exerted profound anti-inflammatory effects in this rodent model of lymphoma.
  
  At least in my humble opinion that does not change the fact that your best sources of vitamin E are natural and that's smart to stay away from alpha-tocopherol only supplements and prefer a whole spectrum tocopherol + tocotrienol supplement. Remember: More is not better, when the one thing that counts are the ratios (read more about vitamin E)!
  

That's it for this week's installment of On Short Notice. As mentioned in the introduction, there are more and even shorter news on the SuppVersity Facebook Wall, which usually gets updated 3+ times per day with I would guess 9-12 items total - depending on whether it's a slow news day or not and the time I have to skim studies and popular science mags and repost summaries and links of and to the latter. Let's see what did we have today, already? Ah, yeah: "Review concludes: Just being patented makes Kinesio® no better than conventional taping" (more), "Interesting stupid science finds that GABA directly mediates energy expenditure." (more), or maybe you are interested in "If rapamycin blocks seizures in model of epilepsy. Is mTOR to blame for both?" (more)?

References:

• Goncharova ND, Vengerin AA, Chigarova OA. Repeated Moderate Stress Stimulates the Production of Dehydroepiandrosterone Sulfate (DHEAS) and Reduces Corticosteroid Imbalance in Old Macaca Mulatta.  Bulletin of Experimental Biology and Medicine Volume 153, Number 5 (2012), 750-753. 
• Lean ME, James WP, Jennings G, Trayhurn P. Brown adipose tissue in patients with phaeochromocytoma. Int J Obes. 1986;10(3):219-27. 
• Lennartsson AK, Kushnir MM, Bergquist J, Jonsdottir IH. DHEA and DHEA-S response to acute psychosocial stress in healthy men and women. Biol Psychol. 2012 May;90(2):143-9.  
• Li WJ, Tang LP, Xiong Y, Zhou XD, Wu LD. The chondroprotective effects of dehydroepiandrosterone probably exerted by its conversion to estradiol. J Steroid Biochem Mol Biol. 2012 Oct 18. 
• Liu J, Shimizu K, Konishi F, Kumamoto S, Kondo R. The anti-androgen effect of ganoderol B isolated from the fruiting body of Ganoderma lucidum. Bioorg Med Chem. 2007 Jul 15;15(14):4966-72. 
• Merlo S, Sortino MA. Estrogen activates matrix metalloproteinases-2 and -9 to increase beta amyloid degradation. Mol Cell Neurosci. 2012 Apr;49(4):423-9.
• Ríos JL, Andújar I, Recio MC, Giner RM. Lanostanoids from Fungi: A Group of Potential Anticancer Compounds. J Nat Prod. 2012 Oct 23.
• Sharma R, Vinayak M. α-Tocopherol prevents lymphoma by improving antioxidant defence system of mice. Mol Biol Rep. 2012 Oct 14.
• Sliva, D.; Labarrere, C.; Slivova, V.; Sedlak, M.; Lloyd, F. P., Jr.; Ho, N. W. Biochem. Biophys. Res. Commun. 2002, 298, 603– 612.
• Unsal Y, Kıvılcım G, Ayşegül A, Arzu A, Esra G, Ercan D, Ayşe S. Matrix metalloproteinase-7 and matrix metalloproteinase-9 in pediatric multiple sclerosis. Pediatr Neurol. 2012 Sep;47(3):171-6.
• Wang KC, Lin YF, Qin CH, Chen TL, Chen CH. Bisphenol-A interferes with estradiol-mediated protection in osteoarthritic chondrocytes. Toxicol Lett. 2010 Oct 5;198(2):127-33. 
• Watanabe M, Ohno S, Nakajin S. Effects of bisphenol A on the expression of cytochrome P450 aromatase (CYP19) in human fetal osteoblastic and granulosa cell-like cell lines. Toxicol Lett. 2012 Apr 5;210(1):95-9.

When Rodents Squat, Scientists Gain Insights into How Muscles Grow. IGF-1 Response to Exercise Does Matter - Locally, not Systemically, of Course!

You want to build big wheels? Look no further get yourself the "Squat T-Bar" with integrated 15mA electrical 'motivator' (Aguiar. 2012)

"A rodent study investigating strength workouts?" Yeah, I know it does not sound like that would be in any ways news-worthy, but if you take a look at the image on the right, you will immediately realize: This study is different! Instead of using a treadmill or simply stitching down (or rather up) one of the hindlimbs of the rodents to induce a chronic overload on the other one (don't laugh, many rodent studies have done just that), the study at hand (Aguiar. 2012), which is going to be published in the next issue of the International Journal of Sports Medicine, used a not innovative, but unfortunately largely forgotten (or overlooked?) torturing device that has been developed by Japanese researchers roughly 20 years a ago (Tamaki. 2012).

The rodent torture... ah pardon squat rack ;-)

After being fitted with a canvas jacket in a way that would enable the researchers to limit the twisting and flexion of their torsos (no, that was not a weight lifting belt ;-), the 32 male Wistar rats (80 days old, 250–300 g) were suspended in a standard position on their hind limbs and "encouraged" to exercise by "electrical stimulation [...] that was applied to the rat’s tail through a surface electrode"  (Aguiar. 2012).

Using their neat little toy, the eight researchers from the University Estadual Paulista, in Botucatu, Brazil, were able to submit the rats to a relatively realistic progressive resistance training regimen for either 8 or 12 weeks. Three times per week each rodent had to do 4 sets  of squats for 10-12 repetitions at 65-75% of its individual 1-RM (maximal weight the rodent could handle). During the study period, Aguiar et al. adjusted the weights twice a week to ensure the same training intensity throughout the experiment (something I would highly recommend to anyone of you, as well; try to pack on 1.25lbs - 2.5lbs at least every other week).

Figure 1: Body weight, muscle weight (plantaris, only) and food intake relative to body weight of the control (C8, C12) and trained (T8, T12) rats before and after the 8-week (C8, T8) and 12-week (C12, T12) intervention (data adapted from Aguiar. 2012)

As you can see in figure 1, this minimalist approach to leg training lead to an increase in both body weight and muscle weight that may initially look as if it was strongly linear. You do yet have to be careful about statements like that, because (a) the rodents did gain weight irrespective of whether they were training or not (80 day old rats are still growing!), so comparing the four bars next to each other and saying "yep, linear!" is not feasible, because this would mean linear as in not training for eight weeks < training for 8 weeks < not training for 12 weeks < training for 12 weeks, which is obviously nonsensical. That being said, there is simply (b) insufficient data to say anything about the linearity -- after all, we do have only three data points per group.

"All groups started the experiment with similar body weight. There was a significant increase (p < 0.05) in the body weight of the 4 groups in the resistance training program (C8: 35.5 %; T8: 27.7 %; C12: 46.9 %; and T12: 40.1 %) and final body weights were not significantly (p > 0.05) different between groups. Furthermore, no significant (p > 0.05) differences in the weekly food intakes were observed between the groups."(Aguiar. 2012)

What does yet stick out, is that the obviously age-dependent weight gain in the control groups C8 and C12 did not increase the weight of the plantaris muscle to a weight anywhere near to the muscle weight, the rats in the trained groups achieved.

Muscle gains and strength gains went hand in hand

In the rats who were subject to the three-times-per-week exercise regimen, on the other hand, those increases in muscle size went hand in hand with highly significant improvements in 1-RM squat power; While all groups had begun the training protocol with similar absolute 1-RMs of ~450g (that's about 130% body weight, pre) ...

"[...] training for 8 and 12 weeks promoted a significant (p < 0.05) increase in the RM/BW ratio in the T8 (pre- vs. post-training: 35.7 % increase, p < 0.05) and T12 (pre- vs. post-training: 57.1 % increase, p < 0.05) groups, while no statistical (p > 0.05) difference was observed in their respective control groups." (Aguiar. 2012)

Consequently, the ratio of 1-RM to body weight was 36.1 % and 57.7 % higher in the groups who had been training fot the last 8 or 12 weeks than in the lazy controls and the time-effect yielded another +22% increase in strength in those rodents who trained for 12 and not just 8 weeks.

Figure 2: Strength gains (left) and increases in cross sectional area, as well as intramuscular IGF1, myogenin and myoD expression (data adapted from Aguiar. 2012)

Now you may have heard all that before, what really makes this study stand out, however, is the observation of statistically highly significant correlations of intra-muscular IGF1, myogenin and myoD  mRNA expression, which speaks in favor of my previous hypothesis (read up on that in the Intermittent Thoughts on Building Muscle) that muscle growth is triggered, driven and maintained almost exclusively at a local level.

What are myogenin and myoD? Both are myogenic regulation factors with myogenin actually being part of the myoD family of transcription factors that will make stem cells develop into myocytes (myo D is highest in recently activated satellite cells).

So, when you are looking for "hormonal" (or other pro-anabolic) ghosts (Phillips. 2012), it is imperative to look for them right where the spook, or, in this case, the muscle building magic happens. If you do just that (see figure 2) and correlate the intra-muscular mRNA expression of IGF-1, myogenin and myoD, you will find the "ghostly" explanation for strength and size gains, as well as the confounding structural changes in the architecture of the muscle, with corresponding correlations between the increases in muscle cross-sectional area (CSA) of r = 0.85 (p = 0.0001), r = 0.87 (p = 0.0001) and r = 0.88 (p = 0.0001) for myoD, myogenin and IGF-1, respectively.

Fiber type changes take their time and occur only within the type II spectrum

A neat side-finding, which is actually no news, though, pertains to the fiber-type conversions that took place in response to the exercise regimen. Firstly, the scientists confirmed the notion that these changes occur exclusively within a certain fiber type. In other words, while Aguiar et al. observed conversions from the metabollically more flexible type IIX/D to the highly glycolytic (power) IIA type, no conversions of the highly oxidative type I to type II fibers were observed. And though the results would generally suggest that fiber IIX/D-to-IIA type conversion, as they 

"[...] also appear to occur during endurance training in humans, so that it would [be] reasonable to think that any exercise stimulus (e. g., endurance or strength) that is sufficient in duration and/or intensity can potentially induce conversions within the fast fiber population from type IIX/D to type IIA" (Aguiar. 2012),

the time-frame in the course of which these changes took place -- namely 12 weeks -- would confirm that the common fear of strength and endurance athletes could provoke negative structral adaptations from doing a "cardio" or "strength" workout from time actually is actually unwarranted. Neither will the former turn a powerlifter into a weakling, nor will the latter make a marathon runner "bulky". Both powerlifter and marathoner are on the contrary going to benefit from the conditioning effect and increase in strength, respectively -- not to mention the important effects on overall health both and not as mainstream stupidity will tell you only the powerlifter can derive from, figuratively speaking, "killing some game in the other's territory"

Bottom line: More food for intermittent thoughts on building muscle ;-)

Figure 3: Correlations between acute GH (A), free testosterone (B), IGF-1 (C)  and cortisol (D) responses (area under the curve—AUC) and gains in type II fibre CSA (Burd. 2012).

Eventually, this study is an excellent example of a way to design a rodent study in a way that will render its results actually meaningful. And what's more, in this particularly case these results are not just meaningful, but can also help us to make some sense of a couple of things we have not fully understood / appreciated, as of yet.

What I am particularly thinking about here, is the contrast between the in-vitro effects of IGF-1 and the (more or less absent) real-world effects of the IGF-1 response to exercise (=systemic increase), as it was observed by West and Phillips in a 2012 study. In their well-powered longitudinal study, neither the acute increase in systemic testosterone, nor the exercise induced increases in systemic IGF-1 showed significant correlations with the gains in type II CSA in a cohort (n = 56) of young men in response to 12 weeks of resistance training (West. 2012; see figure 3).

Another interesting finding of the West study was that, contrary to the circulating testosterone and IGF-1 levels, GH and cortisol did show direct correlations with increased muscle cross sectional areas in type II fibers.

And while the former correlation may be explained by the influence of growth hormone (GH) on the local expression of IGF-1 (Hameed . 2004), there is another open question left: How does cortisol actually figure in here? I mean, the chronic elevation / exogenous adminstration of cortisol, has been show to do the exact opposite, i.e. it decreases the local IGF-1 mRNA expression (Inder. 2010).

Figure 4: Graphical summary of what you should have learned Intermittent Thoughts on Building Muscle ... you didn't 'cause you are new to the SuppVersity or simply forgot about it? No problem read the preliminary summary and browse the individual chapters here!

What was missing in the Inder study, however, was the exercise component: Working out does not just exert protective effects against the negative side effects of the provision of exogenous "cortisol" (in this case Dexamethason), as they were observed in the afore referenced study by Inder et al., exercise will also lead to profound increases in local IGF-1 mRNA expression (e.g. +60% in Bamann. 2001), despite the fact that it will also increase the release of the falsely vilified anti-inflammatory glucocorticoid, cortisol... acute vs. chronic, local vs. system, peak values and amplitudes vs. plateaus and AUC values - you got to keep all these contrastive, yet complementary pairs in mind, when you are thinking about the endocrine and intracrine (within the cell) mediators of skeletal muscle hypertrophy.... what? Sounds familiar? Well, you must have been following the Intermittent Thoughts on Building Muscle, then ;-)

References:

• Aguiar AF, Vechetti-Júnior IJ, Alves de Souza RW, Castan EP, Milanezi-Aguiar RC, Padovani CR, Carvalho RF, Silva MD. Myogenin, MyoD and IGF-I Regulate Muscle Mass but not Fiber-type Conversion during Resistance Training in Rats. Int J Sports Med. 2012 Oct 11.
• Bamman MM, Shipp JR, Jiang J, Gower BA, Hunter GR, Goodman A, McLafferty CL Jr, Urban RJ. Mechanical load increases muscle IGF-I and androgen receptor mRNA concentrations in humans. Am J Physiol Endocrinol Metab. 2001.
• Ding H, Gao XL, Hirschberg R, Vadgama JV, Kopple JD. Impaired actions of insulin-like growth factor 1 on protein Synthesis and degradation in skeletal muscle of rats with chronic renal failure. Evidence for a postreceptor defect. J Clin Invest. 1996 Feb 15;97(4):1064-75. 
• Inder WJ, Jang C, Obeyesekere VR, Alford FP. Dexamethasone administration inhibits skeletal muscle expression of the androgen receptor and IGF-1--implications for steroid-induced myopathy. Clin Endocrinol (Oxf). 2010 Jul;73(1):126-32.
• Phillips SM. Strength and hypertrophy with resistance training: chasing a hormonal ghost. Eur J Appl Physiol. 2012 May;112(5):1981-3-
• Sculthorpe N, Solomon AM, Sinanan AC, Bouloux PM, Grace F, Lewis MP. Androgens affect myogenesis in vitro and increase local IGF-1 expression. Med Sci Sports Exerc. 2012 Apr;44(4):610-5.
• Tamaki T, Uchiyama S, Nakano S. A weight-lifting exercise model for inducing hypertrophy in the hindlimb muscles of rats. Med Sci Sports Exerc. 1992 Aug;24(8):881-6.
• West DW, Phillips SM. Associations of exercise-induced hormone profiles and gains in strength and hypertrophy in a large cohort after weight training. Eur J Appl Physiol. 2012 Jul;112(7):2693-702.