Want to Home-Brew Your Own 15x More Bioavailable Super-Curcumin? Buy Buttermilk and a Yogurt Starter Culture

No one says you cannot add other ingredients to the yogurt to make it more tasty if you add the curcumin before fermenting the buttermilk.

If you're a regular at the SuppVersity you will know that curcuminoids, the polyphenols found in turmeric roots (Curcuma longa), have health effects that are similar, in some cases even superior to several anti-inflammatory, anti-diabetic and lipid-lowering drugs. Yes, their consumption has even been linked to significant reductions in cancer risk. Unfortunately, there's a problem with these powerful polyphenols: they are hydrophopbic (Tønnesen. 2002) and prone to degradation in an aqueous environment at neutral and alkaline pH (Tønnesen. 1985; Wang. 1997) - two properties of which Gupta and others (2013) believe that they are responsible their poor oral bioavailability.

Unlike other supps and practices curcumin has not been shown to interfere with hormesis

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Low bioavailability or not, Shishan Fu and rightly highlight in the introduction of their latest paper, even the hardly absorbed "regular" curcuminoids have been reported to offer many health-promoting properties (Gupta and others 2013), it is thus only logical that there's an "interest in the development of functional foods containing these compounds is increasing" (Fu. 2016).

Figure 1: Even dispersing them in buttermilk will increase the bioavailability by decreasing the breakdown of curcuminoids during digestion - that's at least what a 2014 study by Fu et al. shows. 

Hitherto scientists have (more or less successfully) tried to increase the solubility and stability of curcuminoids by dispersing them in matrices such as lipid-based emulsions (Ahmed. 2012; Yu and Huang. 2012), modified starch (Yu and Huang 2010), hydroxypropyl methyl cellulose (Chuah. 2014), milk proteins (Yazdi and Corredig. 2012), and buttermilk (Fu. 2014). As Fu et al. point out, ...

"[...t]he bioavailability may also be increased when formulated in appropriate delivery systems. For example, lecithin–piperine formulations containing curcuminoids and curcuminoids encapsulated in cellulose have been reported to have enhanced bioavailability after oral administration in humans (Antony and others 2008; Vitaglione and others 2012)" (Fu. 2014).

Based on their own previous study with regular buttermilk and evidence that yogurt can significantly increase the stability and bioavailability of bioactives, like green tea polyphenols (Lamothe and others 2014), Fu et al. speculated that dispersing curcuminoids in buttermilk prior to yogurt manufacture would exert even more powerful effects than simply mixing them with buttermilk (see Figure 1). To test this hypothesis, the scientists did something anyone of you can do at home (see Figure 2 for information on how the control samples were prepared, too):

Figure 2: Preparation of yogurts (Fu. 2016).

"A buttermilk dispersion (14% total solids, w/w) was prepared by reconstituting 142.3 g of buttermilk powder in MilliQ-water which was made up to 1000 g. The powder was dispersed in the water at 45 °C with stirring using an overhead stirrer (Heidolph RZR 2051 control, Germany) at 1000 rpm for 30 min. The dispersion was then stored at 4 °C overnight for more complete hydration. The chosen fortification level of curcuminoids in yogurt was 300 mg/ 100 g yogurt (0.3% w/w)

An ABT-5 culture was prepared by mixing 0.2 g of culture granules in 10 mL of buttermilk dispersion (14% total solids, w/w) and stirring for 15 min in an ice bath. This culture solution was prepared freshly prior to fermentation. The ABT-5 culture was added at a level of 0.2 g/L of yogurt buttermilk. The buttermilk was subsampled (50 mL) into separate plastic containers and incubated at 43 °C until pH reached 4.6. These set yogurts were put into the ice water bath for 30 min, stirred at 200 rpm for 20 s using a mixer (Heidolph RZR 2050, Germany) and then stirred manually (approximately 20 times) to obtain a uniform product. The stirred yogurts were stored in a cool room (4 °C) overnight. All analysis was completed within 2 d of yogurt manufacture. The total solids of the yogurts were estimated using a moisture analyser (Sartorius AG, Germany)." (Fu. 2016).

To be able to tightly control the experiment, the curcumin enhanced yogurt and the other samples were exposed to in vitro digestion. During this procedure, the sample (5 g) was mixed with 15 mL of simulated gastric fluid (SGF) containing 2 g NaCl and 7 mL 37% w/v HCl per liter (pH 1.23) and 3.2 mg/mL pepsin, and incubated in a water bath with 100 rpm at 37 °C for 2 h (United States Pharmacopeia Convention 2009).

"After exposure to SGF, the mixture was adjusted to pH 6.5 using 1 M NaOH and mixed with 9.6 mL of simulated intestinal fluid (SIF) containing 3 mL of 2 M NaCl, 0.3 mL of 0.075 M CaCl2, and 6.3 mL of 36.5 mg/mL bile extract in 5 mM phosphate buffer. The pH was adjusted to 6.8 and then 5.4 mL of 10 mg/mL pancreatin in phosphate buffered saline was added. Samples were incubated at 37 °C, 100 rpm for 3 h and then placed in an ice bath to arrest the enzyme activity. At the end of the in vitro digestion period curcuminoids were extracted from the whole digested mixture with acetone and quantified using HPLC-DAD" (Fu. 2016). 

The in-vitro digestion, which is described in a previous paper by Fu et al. (2015), provided the scientists with an estimate of the amount of undegraded curcuminoids - it is yet not a 100% reliable method to determine the real world biological effects in humans, which would have to be tested in future studies. In view of the fact that the scientists calculations show that the resistance of the curcuminoids to degradation after sequential exposure to SGF and SIF improved more than just statistically significantly (see Figure 3), it is logical to assume that benefits would be observed in vivo, too.

Figure 3: Bioaccessibility of curcuminoids after sequential exposure of samples to SGF and SIF (Fu. 2016).

The difference pre-processing, i.e. the prior dissolution in ethanol (which wouldn't make you drunk, anyway, because the total ethanol content of the yogurt would be marginal), the dissolution of curcuminoids in buttermilk and its fermentation to "curcumin enhanced" buttermilk yogurt with a standard ATB yogurt starter culture, made in terms of the bioavailability is after all huge.

In fact, the bioavailability of the curcuminoids increased to an extent that easily surpasses the hyped "super / bio curcumins" that use a combination of curcumin and bioperin, which has been shown to exhibit a 6.93-fold higher bioavailability. In all fairness, we shouldn't forget, though, that, unlike the yogurt trick described here, Biocurcumax™ has already been studies in humans (Antony. 2008).

Curcumin has cancer protective effects, too | learn more

The total bioavailability is still low, but... As, Fu et al. point out in the conclusion of their soon-to-be-published paper, "[t]he most important and practical finding from the bioaccessibility data is that the incorporation of powdered curcuminoids into buttermilk prior to yogurt manufacture results in a 15-fold increase in bioaccessibility of curcuminoids compared to that of neat curcuminoids dispersed in aqueous buffer" (Fu. 2016).

The scientists are yet also right to point out that even with the enhanced bioaccessibility of curcuminoids the total bioavailability was still low (approximately 6%) when they were delivered in yogurt.

In view of the fact that the polyphenols which are transferred into the colon are degraded by gut microflora and the degradation products contribute to the bioactivity of these compounds in the body, the real-world relevance of this astonishing increase in bioavailability will have to be tested in in vivo, before we can have a final say on the practical significance of these findings | Comment!

References:

• Ahmed, Kashif, et al. "Nanoemulsion-and emulsion-based delivery systems for curcumin: encapsulation and release properties." Food Chemistry 132.2 (2012): 799-807.
• Antony, B., et al. "A pilot cross-over study to evaluate human oral bioavailability of BCM-95® CG (Biocurcumax™), a novel bioenhanced preparation of curcumin." Indian journal of pharmaceutical sciences 70.4 (2008): 445.
• Chuah, Ai Mey, et al. "Enhanced bioavailability and bioefficacy of an amorphous solid dispersion of curcumin." Food chemistry 156 (2014): 227-233.
• Fu, Shishan, et al. "Bioaccessibility of curcuminoids in buttermilk in simulated gastrointestinal digestion models." Food chemistry 179 (2015): 52-59.
• Fu, Shishan, e al. "Enhanced Bioaccessibility of Curcuminoids in Buttermilk Yogurt in Comparison to Curcuminoids in Aqueous Dispersions." Journal of Food Science (2016): Ahead of print. doi: 10.1111/1750-3841.13235
• Yazdi, S. Rahimi, and M. Corredig. "Heating of milk alters the binding of curcumin to casein micelles. A fluorescence spectroscopy study." Food Chemistry 132.3 (2012): 1143-1149.
• Yu, Hailong, and Qingrong Huang. "Enhanced in vitro anti-cancer activity of curcumin encapsulated in hydrophobically modified starch." Food Chemistry 119.2 (2010): 669-674.
• Yu, Hailong, and Qingrong Huang. "Improving the oral bioavailability of curcumin using novel organogel-based nanoemulsions." Journal of agricultural and food chemistry 60.21 (2012): 5373-5379.

Recovery Cocktail With Vitamins C+E, Ibuprofen, Cold Water Immersion and Whey Works - Long-Term Effects? Unknown!

What's the right strategy to boost workout recovery? A recent study suggests that it could be a mix of NSAID, antioxidants, cold water and whey.

Vitamin C + E, ibuprofen, cold water immersion and whey? Two of these agents have been shown to impair the adaptational response to exercise and thus potentially compromise long-term gains (vitamin C+E and cold water immersion). The other two are either purported (ibuprofen) or proven (whey) ergogenics - ergogenics of which a recent study by scientists from the Hashemite University in Jordan and the University of Alabama in the USA shows that their combination with the formerly named agents is "helpful in protecting performance" in a test during which the competitive athletes (current or former Division I college athletes / club athletes) performed two bouts of high-intensity anaerobic cycling separated by 30 minutes of rest.

Learn more about hormesis and how antioxidants can also impair your gains

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The participants had been randomly assigned in counterbalanced order to start the first week as a treatment or as a control (nontreatment).

• In the treatment trial, participants were provided with 2 oral doses of 1,000 mg of vitamin C (ascorbic acid with citrus bioflavonoids; General Nutrition Corp., Pittsburgh, PA, USA) and 400 IU of vitamin E soft gel capsules (d-a-tocopherol; General Nutrition Corp.). The first dose was taken with dinner the night before the exercise protocol, and the second dose was taken on the morning of the exercise test. The morning doses were consumed at least 1 hour before the blood sample was taken. Ibuprofen doses (400 mg; 2 ADVIL liquigels, 200 mg capsules; Wyeth Consumer Healthcare, Madison, NJ, USA) were given to participants 30 minutes before each exercise session. The protein supplement, 23 g of whey protein (10.6 g essential amino acids [EAA], 7.3 g of conditionally EAA, and 5.6 g of non EAA ON Sunrise, FL, USA), was mixed with 200 ml of skimmed milk to form a protein shake. The protein shake was given to participants within 3 minutes of finishing each exercise session in the treatment trial. Three to 5 minutes after the end of each exercise session, participants submerged their lower body in cold (10–12.58 C) water for 10 minutes.
• In the control trial, the subjects performed the same testing protocol consisting of 2 exercise sessions with 6.5–7 hours between AM and PM sessions to replicate morning and afternoon workouts or heats. 

The exercise tests were 2-bout sessions that began with a 15-minute warm-up (stretching and cycling). Each bout consisted of three 30-second Wingate power tests with 3 minutes of active recovery (60 RPM with no resistance) in between. After the 27th minute, a second warm-up for 3 minutes preceded the second Wingate bout for a total of six 30-second Wingate tests per session (Figure 1).

Figure 1: Representation for the Wingate test daily order (Al-Nawaiseh. 2016).

"All Wingate tests were performed using 7.5% of body weight as a resistance. The resistance was applied to the ergometer (E224 Monark) after a 10-second countdown. Participants used the countdown time to accelerate peddling speed. [...] Participants rated their RPE and muscle soreness sensation (pain) before and after each Wingate test. Muscle soreness was assessed using a 10-cm visual analog scale with anchor points “no pain at all” at the left end and “unbearable pain” at the right end. Rated perceived exertion was determined using a 6–20 point scale" (Al-Nawaiseh. 2016).

When they had volunteered for the study, all participants had been asked to quit any kind of exercise and all kinds of supplements 48 and 72 hours before exercise protocols, respectively.

Intense training sessions will always increase ALT, AST & CK - in some cases to extreme values that are 10-100x above "normal". Unfortunately doctors will never learn that in med-school and may misinterpret these changes as indicators of organ failure | learn more!.

Which markers of recovery are actually useful? A recent study from the Federal University of Uberlandia (UFU) in Brazil says (Bessa. 2016): "The best way to use biomarkers to monitor athletes is to perform a screening test like the test we have performed in this experiment, using the type and intensity of exercise commonly practiced by the athletes" (Bessa. 2016). Which test? Well, the study investigated CK, LDH, cardiac troponin T (cTnT), g-glutamyltransferase (gGT), and C-reactive protein (CRP), interleukin-6 (IL-6), MCP-1, and tumor necrosis factor (TNF)-alpha, the neutrophil to lymphocyte ratio, GPX, SOD, CAT, TAS and TBARS immediately before and 3, 6, 12, 24, 48, and 72 hours after exercise. The parameters the scientists consider most useful, however, were: (1) CK (not LDH) plus a differential analysis based on cTnT (cardiac CK), gGT (liver), hematocrit, and platelet levels that is used to make sure the increase in CK is coming solely from muscle damage and (2) the leukocyte to neutrophil ratio as a marker of the progress of the supercompensation process after workouts.

Accordingly, we can expect that the improved mean wattage in the supplement trial is the result of the wicked mix of antioxidants, pain killers, cold water immersion and whey the subjects had to stomach / endure.

Figure 1: The treatment (orange) ameliorated the decrease in mean and minimum power during the PM Wingate trial that was performed after the ingestion of antioxidants, ibuprofen, and whey and cold water immersion (Al-Nawaiseh. 2016).

What is quite surprising about these improvements is that they occured in the absence of reduced perceived pain scores / fatigue during the PM sessions. In conjunction with the lack of significant effects on muscle CK in the blood (not shown in Figure 2), this observation warrants the authors conclusion that the observed effects on the subjects' performance were "apparently not due to reduced muscle soreness or damage" (Al-Nawaiseh. 2016). That's in contarst to previous studies like

• Pizza et al. (1999), who reported that similar ibuprofen doses lowered CK activity relative to a placebo 3 days after eccentric arm exercise, or
• Tokmakidis et al. (2003), who found that ibuprofen doses (400 mg every 8 hours for 48 hours) did lower the non-athletic subjects' CK levels and reduced their muscle soreness, without, however, helping to restore muscle function compared with placebo.

Whether these difference are the mere result of differences in the way the exercise / tests were timed (vs. Pizza et al.) or differences in the training level of the subjects, as well as the exercise and testing protocols (vs. Tokmakidis et al.) will have to be tested in future studies.

Bottom line: As the scientists points out in the conclusion to their study, their results "suggest that although the combined use of ibuprofen, cryotherapy, vitamins C and E, and protein drink did not significantly help in protecting from muscle damage and soreness, the combination did help in restoring important muscle function and boosted short-term recovery from high-intensity anaerobic performance" (Al-Nawaiseh. 2016).

Using Ice / Cold Water Immersion After Workouts Will Impair Muscle and Strength Gains, as well as Vascular Adaptations | more

Practically speaking, athletes who compete in events that require repeated anaerobic performance (over 24h) should thus benefit from the combined use of antioxidant vitamins, an NSAID (ibuprofen), 10 minutes of lower body cold water submersion, and 23 g whey protein (10 g EAA).

What you should not forget, however, is the fact that at least 2 of the 4 'ingredients' of this 'recovery cocktail' have been shown to inhibit the long(er)-term adaptation to exercise. The chronic use of cold water immersion and high(er) doses of vitamin C + E in conjunction with exercise can thus not be recommended... and if you care about organ and cartilage health, the same goes for the use of NAIDs | Comment!

References:

• Al-Nawaiseh, Ali M., Robert C. Pritchett, And Philip A. Bishop. "Enhancing Short-Term Recovery After High Intensity Anaerobic Exercise." The Journal Of Strength & Conditioning Research (2015).
• Bessa, Artur, et al. "Exercise intensity and recovery: Biomarkers of injury, inflammation and oxidative stress." J. Strength Cond. Res (2016).
• Pizza, F. X., et al. "Anti-inflammatory doses of ibuprofen: effect on neutrophils and exercise-induced muscle injury." International journal of sports medicine 20.2 (1999): 98-102.
• Toakmatidis, Savvas P., et al. "The effects of ibuprofen on delayed muscle soreness and muscular performance after eccentric exercise." The Journal of Strength & Conditioning Research 17.1 (2003): 53-59.

Ramadan Fasting Studies Showing Fat, but no Muscle Loss Support Benefits of 'Lean Gains'-Style Intermittent Fasting

Remember, Ramadan fasting is not about eating healthy or dieting, after sundown most Muslims consume at least as much energy as on a non-fasting day.

While scientists usually refer to alternate-day fasting as "intermittent fasting", the average fitness enthusiasts will think of Martin Berkhan's "lean gains" protocol, when he or she hears the words "intermittent fasting" - a protocol that involves fasting for minimally 16h and eating for maximally 8h and is thus somewhat similar to the "eat only after sundown" protocol Muslims follow during Ramadan.

Against that background, it makes sense to assume that the two dozen of peer-reviewed Ramadan fasting studies from the Middle East and the Muslim part of Asia provide an (albeit often uncontrolled) model for intermittent fasting.

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If we assume that this premise is true, a recent study from the Dr. Cipto Mangunkusumo General Hospital at the Universitas Indonesia, provides intriguing insights into the lean mass conserving effects of "intermittent fasting".

The study aimed to evaluate the effect of Ramadan fasting on body composition in healthy Indonesian medical staff. To this ends, Ari Fahrial Syam et al. (2016) recorded the body composition of healthy medical staff  members of the Dr. Cipto Mangunkusumo General Hospital before, during and after Ramadan fasting in 2013 (August to October).

Figure 1: Changes in body composition during the 28-day Ramadan fast (Syam. 2016).

In conjunction with data on the energy intake of the forty-three subjects, the data the scientists gathered appear to confirm what the proponents and followers of the "lean gains" variety of intermittent fasting say: You will lose body fat, but not muscle and that without significant reductions in energy intake.

According to 24h food recalls, the subjects didn't change their energy intake during Ramadan. The fat loss can thus not be explained by the induction of a caloric deficit.

Yes, the body composition was assessed by BIA, but ... while body impedance analysis (BIA) may not the best method to assess the %fa and % lean mas of trained athletes, but has been shown to have a high accuracy and reliability in "normal" people. Similarly, the 24-hour food recalls that were used to evaluate the subjects' dietary intake are often a bit off, but that's the case for both pre- and post-assessments, which is why it is reasonable to assume that there was indeed no significant change in total energy intake. This doesn't change, though that it's a pity that the scientists don't provide information about individual nutrients intakes. Previous studies suggest increased protein intakes during Ramadan fasting (Frost. 1987; El Ati. 1995) - a potentially highly relevant increase, obviously.

One thing Syam's study adds to the table, however, is that (a) the weight you will lose is not just body fat, it's also a lot of water and even bone (see Figure 1) and that (b) your (fat) weight will bounce back, within 4-5 weeks when you return to your usual eating habits.

Table 1: Overview of pertinent research comparing the results of the study at hand to previous studies (Syam. 2016).

As the research overview in Table 1 shows, some, but not all of the (side) effects are actual news. Hossini et al. (2013), for example, didn't observe changes in mineral or water content. The initially highlighted lack of protein / muscle loss, on the other hand, was observed in all three studies in which the exact body composition was measured. It is thus not unreasonable to assume that this is a characteristic feature of pertinent "intermittent fasting" protocols.

Fears about fat gain are likewise unwarranted | learn more

Bottom line: Don't get all excited, even if the 24h food recall were accurate and the fat loss occurred in the absence of reductions in food intake, the fat loss was statistically significant, but with less than 500g practically negligible.

The more important message of the study at hand is thus that intermittent fasting doesn't trigger a loss of muscle mass - a fear that is still prevalent, especially in male members of the fitness community | Comment!

References:

• El Ati, Jalila, C. H. I. R. A. Z. Beji, and J. A. B. E. R. Danguir. "Increased fat oxidation during Ramadan fasting in healthy women: an adaptative mechanism for body-weight maintenance." The American journal of clinical nutrition 62.2 (1995): 302-307.
• Frost, G., and S. Pirani. "Meal frequency and nutritional intake during Ramadan: a pilot study." Human nutrition. Applied nutrition 41.1 (1987): 47-50.
• Hosseini, Seyyed Reza Attarzadeh, et al. "The effect of ramadan fasting and physical activity on body composition, serum osmolarity levels and some parameters of electrolytes in females." International Journal of Endocrinology and Metabolism 11.2 (2013): 88.
• Faris Mohammed Ahmed, et al. "Impact of Ramadan Intermittent Fasting on Oxidative Stress Measured by Urinary 15-F2t-Isoprostane." Journal of Nutrition and Metabolism 2012 (2012).
• Norouzy, A., et al. "Effect of fasting In Ramadan on body composition and nutritional intake: a prospective study." Journal of Human Nutrition and Dietetics 26.s1 (2013): 97-104.
• Sadiya, Amena, et al. "Effect of Ramadan fasting on metabolic markers, body composition, and dietary intake in Emiratis of Ajman (UAE) with metabolic syndrome." Diabetes, metabolic syndrome and obesity: targets and therapy 4 (2011): 409.
• Syam, Ari Fahrial, et al. "Ramadan Fasting Decreases Body Fat but Not Protein Mass." International Journal of Endocrinology and Metabolism 14.1 (2016).

Minimal Amounts of Fish Peptide Hydrolysate Double Fat Loss Compared to Whey Isolate on Energy Restricted Diet

I certainly recommend eating fish. Whether I will be recommending fish hydrolysate supplements in the future, however, will have to be determined when additional studies with different baseline diets will have been published.

You may remember that I've written about fish protein hydrolysates / peptides before. Unlike today's article, however, previous articles dealt with the effects of fish protein in rodents. Intrigued by in vitro and animal studies showing that fish-derived peptides demonstrated antihypertensive (Hatanaka. 2009; Kim. 2012; Li. 2012; Ngo. 2011), antioxidant (Nazeer. 2012; Najafian. 2012), immunomodulating effects (Duarte. 2006), reparative properties in the intestine (Fitzgerald. 2005; Marchbank. 2008), and effects in reducing plasma cholesterol and triglycerides levels (Möller. 2008), a group of Italian researchers decided to investigated the effect of Slimpro(R), a supplement containing commercially available fish protein hydrolysate from blue whiting (Micromesistius poutassou), on body composition and on stimulating cholecystokinin (CCK) and glucagon-like peptide-1 (GLP-1) secretion in 120, overweight, non-obese (25 kg/m² < BMI < 30 kg/m²), male (25%) and female (75%) subjects aged 18 - 55 year.

Do not underestimate fish as a protein source - fish is more than just omega-3!

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Unlike the product that was used may suggest, the study was not sponsored by the supplement company. The authors received neither funding nor other external support and they also declare that they don't have a conflict of interest that may be related to patents or direct involvements in the industry. I guess it's important to point that out, even though fact that the scientists chose whey, i.e. an actually relevant control, instead of carbohydrates or just plain water, may have given away the lack of sponsorship, anyways.

Two weeks before the study started, subjects were asked to fill in an alimentary diary reporting their food preferences. A mild hypocaloric ( 300 kcal/day) diet was elaborated for each subject by a dietitian based on subject’s food preferences and habits as reported in the alimentary diary.

Figure 1: The low protein content of the diet is - as highlighted in the annotations to this graphical illustration of the macronutrient composition of the test diets - problematic, to say the least.

Approximately, 55% of energy intake was from carbohydrates, 25% from lipids, and the remaining 20% from proteins. Part of these 20% of protein were either 1.4g and 2.8g of fish protein or 1.4g of whey protein isolate as a control (I just assume that the dosage was 1.4g, because there was only one whey group), which were consumed in form of a flavored shake according to the following protocol:

"Both the active (one dose treatment arm) and the placebo products were taken as follows: ‘dilute the content of one sachet in a large glass of cool water (200 ml). Shake or stir with a spoon. Consume within 10 30 min before the main meal’. In the case of two-dose treatment arm, one sachet of the active product was taken 30 min before lunch and one sachet 30 min before dinner" (Nobile. 2016).

To be able to tell what could be responsible for advantages or disadvantages of the two treatments, the scientists assessed more than just body weight, fat mass (DXA scans), and safety of use as well as the secondary efficacy endpoints, extracellular water, and the circumference of waist, hips, and thighs. They also checked the CCK and GLP-1 levels in their subjects' blood. This is relevant, because this is how the fish hydrolysate is advertised on the manufacturers website:

"Taken daily before meals, Slimpro® increases the production of CCK and GLP-1 in the body, thus amplifying messages associated with a decrease of food intake. Promising results were reported from in vivo et in vitro trials of these molecules that may control food intake. Scientists have described this ingredient as a direct action on the hunger process" (Nobile. 2015).

As it is usually the case in studies like this, some patients were "lost". In this case, we're talking about a total count of eleven subjects who did not reappear for the follow-up check (One subject in the one-dose treatment arm, four subjects in the twodose treatment arm, and six subjects in the placebo treatment arm discontinued intervention because they were no longer interested to participate in the study). The results of the other subjects are plotted in Figure 2:

Figure 2: Changes in body composition after 45 and 90 days of dieting w/ the specific supplements (Nobile. 2016).

As you can see, double-dosing had astonishingly little effect on the subjects' ability to lose body fat. That's in contrast to switching from fish protein hydrolysate to whey protein isolate, which produced measurably, but not statistically reduced rates of fat loss and waist reductions.

Figure 3: Blood biomarker levels. (a) CCK blood levels and (b) GLP-1 blood levels. Intragroup (vs. D0) statistical analysis is reported upon the bars of the histogram. The lines report the intergroup (vs. placebo) statistical analysis. Statistical analysis is reported as follows: *p < 0.05, **p < 0.01, and ***p < 0.001. Data are mean +/- SE (Nobile. 2016).

And guess what: Even though the bars don't look like it, the asterisks over the bars tell you that these differences may be caused by the same differential expression of the satiety hormones CCK and GLP-1 in the fish hydrolysate vs. whey protein group that has been observed with other control protein in previous studies and is boldly advertised on the producer's website.

Great! Let's eat more fish... It stands out of question that the former is actually a very good idea (assuming you make the right fish choices). I have to warn you, though: Firstly, the fish protein consumed in the study at hand came from fish, but just like whey protein and milk, fish and fish protein hydrolysates will also have different effects.

Is Wild Caught Fish Always the Better Choice? With Sign. More N3 and Less Pollutants?  Learn more!

What is probably way more important, however, is the relative protein deficiency of the subjects. With only 20% of the diet being protein, the study participants hovered around at the meager level of the RDA. Since the effects of 1.4g of fish protein hydrolysate you throw on top of a low protein diet are probably very different from those of the same amount of fish protein consumed alongside 2g/kg of dietary and supplemental protein, I wouldn't guarantee and in fact even doubt that you would see a similar almost 100% increase in fat loss while dieting - and still,  the CCK and GLP-1 boosting effects of fish protein hydrolysates are intriguing | Comment on Facebook!

References:

• Duarte, Jairo, et al. "Immunomodulating capacity of commercial fish protein hydrolysate for diet supplementation." Immunobiology 211.5 (2006): 341-350.
• Hatanaka, Akimasa, et al. "Isolation and identification of antihypertensive peptides from antarctic krill tail meat hydrolysate." Journal of food science 74.4 (2009): H116-H120.
• Kim, Se-Kwon, Dai-Hung Ngo, and Thanh-Sang Vo. "Marine fish-derived bioactive peptides as potential antihypertensive agents." Adv Food Nutr Res 65 (2012): 249-260.
• Li, Ying, et al. "Purification of a novel angiotensin I-converting enzyme (ACE) inhibitory peptide with an antihypertensive effect from loach (Misgurnus anguillicaudatus)." Journal of agricultural and food chemistry 60.5 (2012): 1320-1325.
• Marchbank, T., et al. "Clinical trial: protective effect of a commercial fish protein hydrolysate against indomethacin (NSAID)‐induced small intestinal injury." Alimentary pharmacology & therapeutics 28.6 (2008): 799-804.
• Möller, Niels Peter, et al. "Bioactive peptides and proteins from foods: indication for health effects." European journal of nutrition 47.4 (2008): 171-182.
• Nazeer, R. A., NS Sampath Kumar, and R. Jai Ganesh. "In vitro and in vivo studies on the antioxidant activity of fish peptide isolated from the croaker (Otolithes ruber) muscle protein hydrolysate." Peptides 35.2 (2012): 261-268.
• Najafian, L., and Abd Salam Babji. "A review of fish-derived antioxidant and antimicrobial peptides: their production, assessment, and applications." Peptides 33.1 (2012): 178-185.
• Ngo, Dai-Hung, et al. "Free radical scavenging and angiotensin-I converting enzyme inhibitory peptides from Pacific cod (Gadus macrocephalus) skin gelatin." International journal of biological macromolecules 49.5 (2011): 1110-1116.

High Dose NSAID Boosts Muscle Gains in Elderly Men - 11% Increase in Type II Fiber Size, Type I Grew Only 'on' Tylenol

Are NSAIDs over-the-counter anabolics from the pharmacy next door?

Even though this is not the first SuppVersity article about the effects of NSAIDs or COX-inhibitors like Aspirin, Tylenol, Pain-Eze and co., I would like to highlight one again that the existing evidence suggests differential effects in young(er) vs. old(er) individuals, with the former seeing no or detrimental and the latter no or beneficial effects when using NSAIDs during resistance training regimen.

It is thus neither guaranteed, nor likely that a young man or woman would see the same 28% extra-increase in type I fiber and 11% extra-increase in type II fiber diameter, Trappe et al. describe in their soon-to-be-published paper in the journal of the Gerontological Society of America (Trappe. 2016).

The link to hormesis research is far from being straight-forward

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I do understand, though that the numbers still got your attention. Well, let's take a close look at how the researchers got to these impressive results. It all started with previous research that suggested that common cyclooxygenase (COX)-inhibiting drugs enhance resistance exercise induced muscle mass and strength gains in older individuals.

Unfortunately, the results of the few studies we have, are conflicting (Schoenfeld. 2012; see Table 1) - with one showing benefits and two showing no effect at all. The purpose of Trappe's latest study was thus to (a) simply gather more evidence and (b) investigate the mechanism behind the changes that were observed in previous studies. Or, as the scientists put it "whether the underlying mechanism regulating this effect was specific to Type I or Type II muscle fibers" (Trappe. 2016).

Table 1: Summary of human studies investigating the effect of nonsteroidal anti-inflammatory
drug consumption on muscle hypertrophy (Schoenfeld. 2012).

To this ends, the scientists obtained muscle biopsies from the vastus lateralis of healthy older men who consumed either a placebo (n = 8; 64±2 years) or COX inhibitor (acetaminophen, 4 gram/day; n = 7; 64±1 years | compliance was monitored by researchers, when tablets were taken at the lab or camera when taken at home) during a standardized 12 weeks resistance training program (only the knee-extensor was trained - albeit on 3 days/week) the scientists describe as follows:

"All participants completed a progressive resistance exercise training program of bilateral knee extension that was designed to hypertrophy and strengthen the m. quadriceps femoris, using a protocol employed for several previous investigations in our laboratory. Each participant was scheduled for resistance training three times per week over the 12 weeks for a total of 36 sessions on an isotonic knee extension device (Cybex Eagle, Medway, MA). All sessions were supervised by a member of the research team. Each session was separated by at least 1 day and consisted of 5 minutes of light cycling (828E, Monark Exercise AB, Vansbro, Sweden), two sets of five knee extensions at a light weight, followed by three sets of 10 repetitions with 2 minutes of rest between sets. Training intensity was based on each individual’s one repetition maximum (1RM) and adjusted during the training based on each individual’s training session per formance and biweekly 1RM" (Trappe. 2016).

The compliance of the subjects of this double-blinded study is described as excellent. Therefore, we can assume that the significance of the results of the scientists' analysis of muscle samples that were examined for Type I and II fiber cross-sectional area, capillarization, and metabolic enzyme activities (glycogen phosphorylase, citrate synthase, β-hydroxyacyl-CoA-dehydrogenase) is high.

Figure 1: Pre-/post comparison on fiber (according to fiber type) and muscle size (Trappe. 2016).

Obviously, the most important results of these analysis have been mentioned before: While the type I fiber size did not change with training in the placebo group (304±590 μm²), it increased by a statistically significant and practically relevant 28% in the COX inhibitor group (1,388±760 μm²).

Schematic of the prostaglandin (PG) producing cyclooxygenase (COX) pathway and specific receptors that influence growth and atrophy in skeletal muscle (Trappe. 2013b).

How do COX inhibitors promote hypertrophy? As Trappe et al. point out "[e]vidence from the larger cohort suggests that the augmented muscle growth was primarily mediated by a reduction in intramuscular PGE2 and resultant PGE2 receptor downstream signaling effects (Standley. 2013; Trappe. 2013a,b). Specifically, the COX inhibitor appeared to reduce the negative effects of PGE2 on protein synthesis and degradation, working through established myokines and other cellular regulators of protein turnover. The myocellular findings from the current study suggest that these effects were more pronounced in the Type I fibers, possibly due to a more active PGE2/COX pathway in this fiber type" (Trappe. 2016).

In addition, the authors point out that previous evidence suggests an "additional mechanism for the COX inhibitor–induced supplemental growth, working through PGF2α receptor and protein synthesis upregulation" (Trappe. 2016; referring to Trappe. 2013a,b).

For the type II fibers, both groups recorded significant increases in fiber size. With "only" 26%, the gains of the subjects in the the placebo group (1,432±499 μm2, p < .05) were yet measurably lower than those in the COX inhibitor group whose vastus lateralis type II muscle fiber size increased by 37% (1,825±400 μm², p < .05). In view of the overall benefits the COX group saw, it is thus hardly surprising that the subjects consuming the COX inhibitor recorded significantly greater total muscle CSA gains (see Figure 1, right | note: only the total mass gain was sign. different between groups).

Figure 2: Change in fiber type–independent (A) and fiber type–specific (B) muscle capillarization from the beginning to the end of the 12-week resistance exercise training and drug interventions. CCEF = capillaries in contact with each fiber; CSA = cross-sectional area. *p < .05 vs pre. **p < .1 vs pre.

While enzyme activity (not shown in Figure 2) and capillarization were generally maintained in the placebo group, the capillary to fiber ratio of the subjects in the COX inhibitor group increased by an albeit only borderline significant 24% (p < .1). The citrate synthase activity, on the other hand, increased statistically significantly, but by "only" 18% (p < .05). These differential changes in citrate synthase (important for fat oxidation and endurance) and muscle capillarization further underline the beneficial effects of NSAIDs on the adapatational response to exercise in the elderly.

Figure 2: Two out of three studies find that NSAIDs blunt the satellite cell response to resistance training young people | A: Number of Pax7 cells expressed per number of myonuclei (in %) in muscle biopsies (vastus lateralis muscles) obtained before (pre) and 8 days after maximal eccentric exercise (no block and NSAID); B: Immunohistochemical staining with the use of Pax7 antibody to identify satellite cells on a muscle cross-section (7 m) taken 8 days after exercise (from Mikkelsen. 2009).

Bottom line: There's no reason to doubt the scientists' conclusion that "COX inhibitor consumption during resistance exercise in older individuals enhances myocellular growth, and this effect is more pronounced in Type I muscle fibers" (Trappe. 2016). It is important however, that their results apply only to healthy elderly individuals.

Why only in the elderly? Well based on previous research, there's in fact good reason to doubt that similar benefits would have been observed in younger individuals. The hitherto published results in young people are mixed. A possible explanation for that would be the previously observed "impairment of satellite cell activity" (Schoenfeld. 2012) in response to chronic NSAID consumption - a side effect that may turn out to be detrimental in the long(er)-term, because unlike older individuals, in whom the satellite cell function is compromised, already (Thornell. 2011), young people's long-term gains appear to rely on the myostatin lowering recruitement of additional myonuclei.

Overall, the potential negative effects on satellite cell activity and thus long-term muscle growth, the lack of convincing evidence of benefits in younger individuals and, for young and old alike, the negative side effects of chronic NSAID use on your tendons, gut, kidney and other organs are three good reasons I certainly don't advise to seriously consider "supplementing" NSAIDs daily to augment your muscle gains | Comment on Facebook!

References:

• Mikkelsen, U. R., et al. "Local NSAID infusion inhibits satellite cell proliferation in human skeletal muscle after eccentric exercise." Journal of applied physiology 107.5 (2009): 1600-1611.
• Schoenfeld, Brad J. "The Use of Nonsteroidal anti-inflammatory drugs for exercise-induced muscle damage." Sports medicine 42.12 (2012): 1017-1028.
• Standley, R. A., et al. "Prostaglandin E 2 induces transcription of skeletal muscle mass regulators interleukin-6 and muscle RING finger-1 in humans." Prostaglandins, Leukotrienes and Essential Fatty Acids (PLEFA) 88.5 (2013): 361-364.
• Trappe, Todd A., and Sophia Z. Liu. "Effects of prostaglandins and COX-inhibiting drugs on skeletal muscle adaptations to exercise." Journal of Applied Physiology 115.6 (2013a): 909-919.
• Trappe, Todd A., et al. "Prostaglandin and myokine involvement in the cyclooxygenase-inhibiting drug enhancement of skeletal muscle adaptations to resistance exercise in older adults." American Journal of Physiology-Regulatory, Integrative and Comparative Physiology 304.3 (2013b): R198-R205.
• Trappe, Todd A., et al. "COX Inhibitor Influence on Skeletal Muscle Fiber Size and Metabolic Adaptations to Resistance Exercise in Older Adults." J Gerontol A Biol Sci Med Sci (2016): Advance Access publication January 27, 2016.
• Thornell, Lars-Eric. "Sarcopenic obesity: satellite cells in the aging muscle." Current Opinion in Clinical Nutrition & Metabolic Care 14.1 (2011): 22-27.