May I Salt & Roast My Nuts? Plus: If Catechins Boost One's Energy Expenditure by 400% Why Don't They Work for Me?

One article, two questions, two science-based answers.

If you're like me I bet that you've been asking yourself previously, whether the cheap roasted nuts at the supermarket have the same health benefits as the expensive "raw" nuts from the health-food store... guess what: a recent study by scientists from the Singapore Institute for Clinical Sciences and the University of Otago confirms: "Dry roasting and lightly salting nuts do[es] not appear to negate the cardioprotective effects observed with raw nut consumption, and both forms of nuts are resistant to [dietary] monotony" and thus similarly recommendable health-foods.

Along with the latest "green tea for thermogenesis"-study, which finally answers the important question "If Catechins Boost One's Energy Expenditure by 400% Why Don't They Work for Me?", the Chinese-New-Zealand co-production is one of the two studies in today's SuppVersity article.

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• You can roast and salt your nuts (all puns intended) and still see health benefits (Tey 2016): Unless you're like the average seventh-grader and misunderstand the previous statement, roasted and salted nuts display no health risk whatsoever.
  
  In fact, Siew Ling Tey and colleagues were recently able to show in a randomized crossover study with 72 perfectly healthy participants, who were asked to consume 30 g/day of either raw or dry roasted, lightly salted hazelnuts for 28 days, each that the CVD risk factors the scientists assessed still improved significantly (sign. changes in body composition were not recorded - just for the record: body fat declined by 100-200g, muscle mass increased by ca. 100g).
  

  

  Figure 1: Changes in biochemical parameters in the two groups; only the change in triglycerides (trigs) showed a probably random, yet statistically significant inter-group difference (in favor of the roasted nuts | Tey 2016)

  Furthermore, neither the "raw" nor the "roasted nut"-group saw a decline in their "desire to consume" and "overall liking" of the hazelnuts, which constitute, due to their relatively high omega-6 and extremely high PUFA content, an excellent study object for the potentially negative effects of roasting (one would expect a potential oxidation of PUFAs and downstream ill health effects). Moreover, studies "examining the health effects of consuming hazelnuts are", as the authors point out "relatively sparse despite the fact that hazelnuts are the second most common nut produced worldwide" (Tey 2016).
  
  One downside of roasted nuts I do not want to withhold, though, is the slight decrease in alpha-tocopherol during the roasting process, of which the results of the study do, however, indicate that it had no health-relevant consequences.
• Human study confirms: Green tea + caffeine set your brown fat on fire... assuming that you do have functional brown fat (Yoneshiro 2017) -- In rodents, it's not news that the combination of catechins and caffeine will promote brown adipose tissue thermogenesis. In humans, however, the effect has rarely been observed directly and is, as Yoneshiro et al. point out, "poorly understood".
  
  Reason enough for the Japanese scientists to recruit 15 healthy male volunteers, subject them to fluorodeoxyglucosepositron emission tomography and thus assess their BAT activity before and after a single oral ingestion of a beverage containing 615 mg catechin and 77 mg caffeine, as well as after the chronic ingestion of the same beverage 2 times/d for 5 wk in 10 of the subjects.
  

  

  Figure 2: (A) Study protocol of the acute trial. (B) Study protocol chronic trial. Both of the trials were single-blinded, randomized, placebo-controlled, season-matched crossover studies (Yoneshiro 2017).

  To complicate, ah... I mean to make the study more comprehensive the scientists also evaluated the cold-induced thermogenesis (CIT) after 2 h of "cold" exposure at cozy 19°C. Both the acute and chronic trials were single-blinded, randomized, placebo-controlled, season-matched crossover studies.
  

  

  Figure 3: Change in energy expenditure (adj. for FFM) after the ingestion of the test beverages (left). Thermogenic effects of the catechin or placebo beverage expressed as iAUC of EE (right | Yoneshiro 2017).

  What the authors found was in line with the observations in rodent studies - the effect size, however, was, as it was to be expected, magnitudes smaller; and, more importantly, occurred only in those subjects who were blessed with highly active brown fat depots:
  • A single ingestion of the catechin beverage increased EE in 9 subjects who had metabolically active BAT (mean ± SEM: +15.24 6 1.48 kcal, P < 0.01) but not in 6 subjects who had negligible activities (mean ± SEM: +3.42 6 2.68 kcal).
  • The ingestion of a placebo beverage containing 82 mg caffeine produced a smaller and comparative EE response in the 2 subject groups.
  The scientists multivariate regression analysis revealed a significant interaction between BAT and catechin on EE (b = 0.496, P = 0.003). In other words: The acute effects on your energy expenditure depend on the presence and activity of brown fat cells (see Figure 4).
  

  

  Figure 4: (A) Representative FDG-PET/CT images of the high- and low-BAT subjects. (B) Quantitative BAT activity as the SUV of the high- (n = 9) and low-BAT (n = 6) subjects (Yoneshiro 2017).

  Unfortunately, these fat cells which are already scarce in normal-weight human beings are quasi-non-existent (or dysfunctional, that's not 100% clear) in those people who'd need the fat-burning effects of green tea the most: the obese.
  
  Accordingly, it is important to point out that the beneficial effects of chronic GTE + caffeine intake on the thermogenic response to "cold" (19°C) exposure in the 10 normal-weight Japanese subjects may (and I would dare say that it "will") turn out poorly in the average obese individual.

This FDG-PET image shows where the average (lean) human has active BAT stores (van der Lans 2014).

So what's the verdict? While it is nice to know that roasting and (lightly) salting doesn't turn a health-food like nuts to poisonous trash, I guess that Yoneshiro's study sends the more important message to the fitness community. After all, they finally demonstrated that the answer to the question why green tea extracts don't work for you could be as simple as this: "That's because you don't have the brown fat that's responsive to the effects of the catechins."

That's huge because it shows that (a) the usefulness of green tea catechin supplements depends on the individuals and that (b) those who need "fat burners" the most, i.e. those who are still obese or struggling with their weight, are least likely to benefit from it.

After all, previous studies clearly indicate that the majority of morbidly obese (and older | Sacks 2013) subjects have virtually no brown fat/adipose tissue (Vijgen 2011 | or if they have it, it doesn't respond even to treatment w/ cold, insulin or ephedrine | Orava 2013; Carey 2013). In conjunction with the observation that there's a clear correlation between having active brown fat and having no weight problems (ibid.), as well as the realization that the activity of brown fat in the obese only increases after weight loss (Vijgen 2012), it is thus not surprising that many catechin users are disappointed | Comment on Facebook!

References:

• Carey, Andrew L., et al. "Ephedrine activates brown adipose tissue in lean but not obese humans." Diabetologia 56.1 (2013): 147-155.
• van der Lans, Anouk AJJ, et al. "Cold-activated brown adipose tissue in human adults: methodological issues." American Journal of Physiology-Regulatory, Integrative and Comparative Physiology 307.2 (2014): R103-R113.
• Orava, Janne, et al. "Blunted metabolic responses to cold and insulin stimulation in brown adipose tissue of obese humans." Obesity 21.11 (2013): 2279-2287.
• Sacks, Harold, and Michael E. Symonds. "Anatomical Locations of Human Brown Adipose Tissue." Diabetes 62.6 (2013): 1783-1790.
• Tey, Siew Ling, et al. "Do dry roasting, lightly salting nuts affect their cardioprotective properties and acceptability?." European journal of nutrition (2016): 1-12.
• Vijgen, Guy HEJ, et al. "Brown adipose tissue in morbidly obese subjects." PloS one 6.2 (2011): e17247.
• Vosselman, Maarten J., Wouter D. van Marken Lichtenbelt, and Patrick Schrauwen. "Energy dissipation in brown adipose tissue: from mice to men." Molecular and cellular endocrinology 379.1 (2013): 43-50.
• Yoneshiro, Takeshi, et al. "Tea catechin and caffeine activate brown adipose tissue and increase cold-induced thermogenic capacity in humans." The American Journal of Clinical Nutrition (2017): ajcn144972.

Fat-Blocker Effect of Tea Catechins Confirmed (?) in Man - Sign. Abdominal Fat Loss (5-8%) in 12 Weeks W/Out Diet

Tea catechins (which can also be found in black and jasmin tea | see Figure 3) can help you keep particularly unhealthy abdominal fat (Després. 2012) at bay.

It is one thing to have in-vitro and rodent data that green tea can inhibit the digestion of dietary fat (reported previously in the SuppVersity Facebook News); it is another thing, however, to have a human study like the one Makoto Kobayashi and colleagues are about to publish in the peer-reviewed scientific journal Food & Function that shows that the "[i]ngestion of a green tea beverage enriched with catechins with a galloyl moiety (THEA-FLAN 90S) during a high-fat meal reduces body fat in moderately obese adults" (Kobayashi. 2015).

Ok, the abdominal fat loss does not, as the previous quote from the conclusion appears to suggest, occur instantly right after you've consumed your first tea w/ a single meal.

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Rather than that, 124 subjects (two of the initially 126 subjects 2 dropped out for personal reasons unrelated to the trial), 53 men, 71 women, who consumed similar, albeit non-standardized diets (see Figure 1 | note: physical activity was also identical) and began the study with body fat levels of ca. 31-35% had to consume the previously mentioned tea beverage that contained tea catechins (250 mL with 215.3 mg green tea catechins containing 211.0 mg green tea catechins with a galloyl moiety) twice or three times daily during mealtimes for 12 weeks, before the significant reduction in body fat became visible.

Figure 1: Macronutrient composition (in g an % of energy) of the non-energy reduced diets the subjects consumed; the values in the left pie chart represent a group average of all three intervention groups. Since the data is based on food records with photographs, it is probably more reliable than in your average diet study (Kobayashi. 2015).

Now, in view of the fact that this is not the first study to demonstrate weight loss effects in overweight subjects consuming green tea or, as in most other studies, green tea extracts, the word "during" and thus the fact that the green tea beverage was consumed with at least two of the three meals per day should be highlighted as a specific feature of the study at hand that is highly relevant to its interpretation.

Figure 2: Detailed analysis of the rel. change in fat area in the abdominal depot of the subjects (Kobayashi. 2015)

It is after all the requirement that the green tea beverage had to be consumed with a (preferable high fat) meal that allows the authors to conclude that the significant fat loss Kobayashi et al. measured by the means of computer tomography predominantly in the abdominal area are the result of an inhibition or slowing of the intestinal fat absorption and thus warrant the conclusion that "the ingestion of green tea beverages enriched with CGM together with high-fat meals may be an effective strategy for reducing body fat in moderately obese adults" (Kobayashi. 2015) - an observation of which I would like to add that the underlying mechanism is not 100% certain.

What about weight and, even more importantly, muscle loss? No, losing lean mass was not an issue in, because weight loss (-0.6 and -0.8% in the low and high dose group, respectively | measured by bio-electrical impedance vs. computer tomography as it was the case for the abdominal fat area) was actually not an issue, either. If you want to measure your success on the scale, green tea is thus not going to be the "diet tool of choice" (unless you use it alongside an energy-reduced diet)... however, if you take into account that the placebo group actually did what the average Westerner does, these days, i.e. gain weight and body fat over the 12-week study period, you may argue that you can still see the results on the scale which could finally stand still after years of displaying subtle, but eventually relevant increases in body weight.

The authors base their conclusion that it is "unlikely that absorbed green tea CGM leads to increased energy expenditure, followed by reduced abdominal body fat area" (Kobayashi. 2015) on two reasonable, but experimentally (in this study) not confirmed assumptions which are that little to no catechins actually made it into the bloodstream, because ...

1. the low caffeine content of the beverage limits the bioavailability of EGCG & co (caffeine enhances its bioavailability | Nakagawa. 2009) and
2. the ingestion of the beverage with a meal has been shown to significantly reduce the bioavailability of green tea catechins in comparison to the fasted state (Chow. 2005).

The assumption that its just a blockade of the digestion of fat becomes even more questionable, if you (re-)read my 2014 article on the carb blocking effects of tea... Well, eventually, though, you may argue that it does not matter if the reduction in abdominal fat was due to thermogenic effects, thermogenic and fat-blocking effects or, as the scientists believe, mediated exclusively an "inhibit[ion] or slowing [of the subjects'] intestinal fat absorption" (Kobayashi. 2015). And let's be honest, I guess you're right. What matters is that there were significant reduction ins abdominal fat (visceral, subcutaneous and total abdominal fat area). Reduction of which the data in Figure 2 tells you that ...

1. 

   Table 1: Catechin composition of the test beverages.

   the fat loss in the abdominal area was dose dependent - even if the differences between the low and high dose group did not reach statistical significance (for the exact catechin composition see Table 1 on the right) - and that 
2. roughly 50% of the benefits were lost within only 5 weeks when the subjects stopped consuming the green tea beverage, even though their diet didn't change at all (in fact, they consumed minimally less energy in the withdrawal phase from week 12-17).

Now, (b) is obviously good news for green tea lovers, but bad news for those who cannot imagine consuming green tea containing beverages "for the rest of their lives".

Green tea forever, it is then!? Well, as usual we have to consider what limits the generalizability of the results. Firstly, we are dealing with a group of people who have more than a few pounds of extra-weight on their hips. An abdominal fat loss of 8% in 12 weeks is thus not impossible, but not exactly likely to be seen in someone who starts at a body fat percentage of 15% or less (which is half what the subjects in the study at hand began with).

Figure 3: Catechin content (mg/10ml) of black, green and jasmine tea prepared from commercial tea w/ different infusion times (Bronner. 1998).

The second thing we have to keep in mind is the beverage itself. As you've previously read, it has been enhanced with catechins with a galloyl moiety (CGMs | EGCG, ECG, GCG, CG). Does this mean that you cannot achieve similar results if you simply drink green tea? Luckily, data from Bronner, et al. (1998) suggests otherwise. As you can see in Figure 3, it would take only 100 ml of commercially available freshly brewed (infusion time 3 min) green tea and even less black tea to achieve similar concentrations of EGCG and the other catechins with a galloyl moiety in your tea. Accordingly, the second obstacle to the gene- relizability of the study is actually irrelevant.

Third- and lastly, there's yet still the fast reversal of the effects which suggests that it is necessary to become a habitual tea drinker to see long-term / lasting benefits of green tea (or as the data in Figure 3 suggests even catechin containing tea in general) on your body weight and, more importantly, body fat you're carrying around | Comment on Facebook!

References:

• Bronner, W. E., and G. R. Beecher. "Method for determining the content of catechins in tea infusions by high-performance liquid chromatography." Journal of Chromatography A 805.1 (1998): 137-142.
• Chow, HH Sherry, et al. "Effects of dosing condition on the oral bioavailability of green tea catechins after single-dose administration of Polyphenon E in healthy individuals." Clinical Cancer Research 11.12 (2005): 4627-4633.
• Després, Jean-Pierre. "Body fat distribution and risk of cardiovascular disease an update." Circulation 126.10 (2012): 1301-1313.
• Kobayashi, Makoto, et al. "Green tea beverages enriched with catechins with a galloyl moiety reduce body fat in moderately obese adults: a randomized double-blind placebo-controlled trial." Food & Function (2016).
• Nakagawa, Kiyotaka, et al. "Effects of co-administration of tea epigallocatechin-3-gallate (EGCG) and caffeine on absorption and metabolism of EGCG in humans." Bioscience, biotechnology, and biochemistry 73.9 (2009): 2014-2017.

Chocolicious Statin: One Week "on" 100g/day of 70% Dark Chocolate Improves Cholesterol Profile and Reduces Waist Circumference in "Skinny Fat" Women

With the average German eating 9-10kg of chocolate per year we should be pretty heart healthy... well, if more than 10g of those were dark chocolate, I suppose.

You know what? I am pretty enervated by the reverberations of the notorious "chocolate makes Nobel Laureates" study. Even now, month after the publication, the media hype and the head-shaking on part of anyone who does have the slightest idea of the difference between association and causation, it's still all over the place.

On my part this has lead to a "desensitization" with respect to the word "chocoloate" in medical journals, so that I had almost missed this ground-braking study from the Department of Neuroscience at the University of Tor Vergata in Rome, the National Institute for Mediterranean Diet and Nutrigenomic in Cosenza and the UOS of Pharmacology, Department of Pharmacology at the University Magna Graecia,Roccelletta di Borgia in Catanzaro, Italy (di Renzo. 2013)

Chocolate for the skinny fat

As if it was not already enough that this is a "non-Nobel-Laureate-infected" study on chocolate consumption, it's also study on skinny fats: 15 women in the age of 20-40 years with a normal BMI, but high total body fat mass (FM) percentage (FM% > 30%) and significantly higher values of proinflammatory cytokines, such as IL-1, IL-6, IL-8 and TNF-α. SuppVersity reader what else can you ask for?

Well, ok.... probably you could ask what the study was all about. So, the intervention was pretty simple:After a dark-chocolate-free (DC) washout period, the subjects received 100g of dark chocolate per day for 7-days (50g in the morning another 50g as an afternoon snack or at dinner).The dark chocolate contained (only) 70% cocoa and was provided as an unrestricted gift from Valrhona, Tain l’Hermitage, France. Of the 70% cacao in it, 42.5% were cacao butter with a fatty acid composition of 64.6% saturated fats, 34% monounsaturated fats and only 4.4% PUFA. The sugar in the chocolate was almost exclusively saccharose.

Chronic diet = skinny fat!? (learn more)

Did you know? In previous studies, di Renzo et al. found that 10% of the Italian women fall into the "skinny fat", or as they put it "normal-weight obese" category. These women had similar increased cardiovascular disease (CVD) risk indexes values, as their pre-obese peers, but did not manifest the metabolic syndrome, despite a cluster of metabolic and genetic features associated with increased CV mortality (di Renzo. 2006, 2009, 2010; Marques-Vidal. 2010).

Moreover, the chocolate contained 2% polyphenols, and 0.7% theobromine, as well as 444mg/kg naturally occurring catechins, 908mg/kg epicatechin and 20, 20 and 40mg/kg epigallocatechin gallate epicatechin gallate, Epigallocatechin, respectively.

Improved blood lipids, improved body composition, improved....

The main outcomes of the study were the changes in body composition (DXA), blood pressure, anthropometric measurements, biochemical parameters and plasma levels of some cytokines within the 7-day intervention period.

Figure 1: Changes in lipid and glucose metabolism during the 7-day chocolate intervention (di Renzo. 2013)

And as you can see in figure 1, the results were ambiguous. We have the great and statistical signifcant improvement on the cholesterol side of things, as well as a reduction in specific markers of infallamtion, i.e. IL-6 (-33%), TNF-α (-19%) and IL-1Ra (-33%) and a 1cm reduction in waist circumferene. On the other hand, we do also see increases in fasting insulin and insulin resistance, which did not reach statistical significance, but remind me of the inverse effects on lipid and glucose metabolism we see with so many other agents with cholesterol lowering effects.

Still if we rely on statistics, the overall effects is a beneficial one, but...

There is another string attached, though, due to which I still cannot wholeheartedly recommend to add those 100g of chocolate to your diet, because the scientists did not have the funds for a control group... that's a major bummer. After all, we don't really know if the changes were not brought about by the standardization of the diet, which took place at the beginning of the DC abstinence period and of which the authors write:

Did you know? In another study 40g of dark chocolate consumed on a daily basis for 2 weeks were able to reduced the urinary excretion of the stress hormone cortisol and catecholamines and partially normalized stress-related differences in energy metabolism in 30 human subjects, who were classified in low and high anxiety traits using validated psychological questionnaires. (Martin. 2009)

"Total daily energy content of the diet was determined on an individual basis, calculated using De Lorenzo et al. prediction equation for the Italian population. Initial caloric levels were adjusted, when necessary, to maintain the body weight. All subjects received about 1700 kilocalories/day. The recommended composition of the dietary regimen was as follows: carbohydrates, 55% to 60%; proteins, 15% to 20% (of which about 50% was comprised of vegetable proteins); total fat, 25% (saturated fat acids (SFA), less than 10%, and cholesterol consumption, less than 300 mg per day), and 30 g of fibre.

[...] The composition of the diet in terms of foods and food combinations was planned to obtain an animal to vegetable protein ratio as close to 1:1 as possible. The Italian Recommended Dietary Allowances were incorporated to ensure proper vitamin and mineral intake."

Otherwise, the subjects were were advised "not to consume any other chocolate for the duration of the study" and to make "no further changes to their diet and lifestyle habits" (di Renzo. 2013).

---

Bottom line: I do not question the value of adding a chunk of dark chocolate to your diet from now to then and I believe the beneficial effects on your blood lipids is well-established enough to even suggest the supplementation with "pure" chocolate (90%+) just for this porpose.

The EDC Program - Exercise + Diet + Chocolate (learn more)

The question is yet, do you need that? If you are skinny fat, probably. If you work out regularly and have your diet in check, unlikely. So, why would you want to run the risk of being among those of the ladies in the study at hand who must have experienced pretty extreme elevations in fasting insulin. If we were talking about something that happened across the board to all of them, the data would be statistical significant. The way it is, my best bet is that the increase in circulating free fattty acids in response to the combination of catechins + theobromine may be to blame. And this response can very well be very different from one individual to the other. But who knows, maybe it was just the dietary switch (?) to an up to 60% carbohydrate diet that was to blame for these results!?

References:

• Di Renzo L, Bigioni M, Bottini FG, Del Gobbo V, Premrov MG, Cianci R, De Lorenzo A. Normal Weight Obese syndrome: role of single nucleotide polymorphism of IL-1 5Ralpha and MTHFR 677C-->T genes in the relationship between body composition and resting metabolic rate. Eur Rev Med Pharmacol Sci. 2006 Sep-Oct;10(5):235-45. 
• Di Renzo L, Gloria-Bottini F, Saccucci P, Bigioni M, Abenavoli L, Gasbarrini G, De Lorenzo A. Role of interleukin-15 receptor alpha polymorphisms in normal weight obese syndrome. Int J Immunopathol Pharmacol. 2009 Jan-Mar;22(1):105-13.
• Di Renzo L, Galvano F, Orlandi C, Bianchi A, Di Giacomo C, La Fauci L, Acquaviva R, De Lorenzo A. Oxidative stress in normal-weight obese syndrome. Obesity (Silver Spring). 2010 Nov;18(11):2125-30.
• Marques-Vidal P, Pécoud A, Hayoz D, Paccaud F, Mooser V, Waeber G, Vollenweider P. Normal weight obesity: relationship with lipids, glycaemic status, liver enzymes and inflammation. Nutr Metab Cardiovasc Dis. 2010 Nov;20(9):669-75.

Probiotics + Green Tea - Synergistic Superstack or Sciency Non-Sense? Green Tea Alone Totally Blunts HFD Induced Weight Gain, L. Plantarum Does Not Add to Its Effects

L. plantarum may metabolize green tea phenols, but don't add to their anti-diabesity effects 

Green tea has actually never seized being all the rage and probiotics are the sexy new kid on the block right around the corner of the supplement shops and and science laboratories of the western hemisphere. Against that background I guess that the title of a paper that's been published ahead of print on Monday will probably suffice to catch your interest: "Green tea powder and Lactobacillus plantarum affect gut microbiota, lipid metabolism and inflammation in high-fat fed C57BL/6J mice." (Axling. 2012) - and that despite the fact that "mice are no little men" ;-)

'1 + 1 =4' the synergism of green tea and probiotics could make it possible

I guess, the idea sounds logic: Take one thing that has been proven to ameliorate diet induced obesity, namely green tea, and combine that with another one, of which it appears as if it would also exhibit beneficial effects into an even more potent stack. In fact, the scientists' rationale was yet slightly different:

"The species Lactobacillus plantarum (L. plantarum) has the ability to metabolize phenolic acids  and to split up tannins. The metabolites are presumably more easily absorbed and distributed into the tissues where they can act as antioxidants and electron scavengers. Phenolic compounds can also have antimicrobial effects that may affect the composition of the gut microbiota, in favour of polyphenol-metabolizing components of the microbiota. Also, green tea extracts have been shown to selectively inhibit the growth of pathogenic bacteria while either enhancing or not affecting the growth of beneficial bacteria like lactic acid bacteria. To the best of our knowledge, the impact of green tea powder as a prebiotic compound to promote lactobacilli or other health promoting components of the microbiota has not previously been evaluated."

In other words, the expected benefits of providing both green tea and probiotics in conjunction were (1) an increased bioavailability of the phenols and tannins from the green tea that would be induced by the probiotics and (2) an increase in the probiotics' survival and ability to modify the gut microbiome that would be brought about by the addition of the green tea.

What looks good on paper does not necessarily work out in a complex organism

Figure 1: Ingredient total amount of Flavan-3-ol, Phenolic acid and Flavenol in water and methanol extracts from the green tea leaves that have been used in the study (Axling. 2012); as you can see the total quantity and the ratios of the bioactive ingredients of the extract actually depend on the extraction method.

Apropos green tea, you can see the exact ingredient profile of the green tea supplement that has been used in the study at hand in figure 1. In view of the fact that the C57BL/6J mice received no extract, but simply powdered green tea leaves, it may not be important in this context, but could be relevant for your future purchases that methanol and water extracts differ not only in terms of the total amount of Flavan-3-ol, Phenolic acid and Flavenol they contain, but also with respect to the ratio of the respective phytochemicals. I guess, those of you who have been around in September 2011, already, will remember that I have discussed the impact these ostensibly negligible differences can have more than a year ago in "-20% Reduction in Serum Testosterone by 5 Cups of Green Tea. Endocrine Effects Depend on Catechin Composition". In case you are one of the many newcomers or have simply forgotten (let alone missed ;-) this post, I suggest you go back and read that up, as it may help you get a better understanding of the underlying reasons due to which quality and quantity of the health effects of green tea (supplements) wary from study to study... but let's now get back to the experimental setup of the Axling study.

Green tea alone already blunts HFD induced weight gain

As mentioned before the extracts were simply mixed with the high fat diet, the mice were consuming in the course of the 22 week study period. With the probiotic supplement that was administered with the drinking water (L. plantarum at 1.5% (v/v) or roughly 3 × 10^9 cfu/ml) we are thus dealing with four different groups:

• Control: High fat chow + no supplement
• LP: High fat chow + L. plantarum
• GT: High fat chow + green tea
• GT + LP: High fat chow + green tea + L. plantarum

If you focus solely on the initially quoted hypothesis about the synergistic effects of green tea + L. plantarum, the actual study outcomes - at least as far as the blood markers in figure 2 are concerned  - are certainly disappointing.

Figure 2: Glucose insulin, fructosamine, cholesterol, triacylglycerol, non-esterified fatty acids and adiponectin levels in the blood of the mice in week 11 and week 22 of the study (Axling. 2012)

It's not like '1+1 would equal 4', but rather like '1 + 1' would just be sufficient to yield '1' not just '0.9' or even less. The in fact, the addition of the probiotics, alone, did very little within the first 11 weeks as far as it's ability to th reduce the diet-induced insulin resistance is concerned and it's addition to the green tea supplement did not improve blood glucose and lipid management, but did in fact diminish the impressive effects the green tea supplement brought about.

Figure 3: Relative change (compared to control) in bacterial diversity and lactobacilli count in response to the supplement regimen (Axling. 2012)

That the probiotic was basically useless, is actually no wonder if you take a closer look at the changes of gut microbiome in figure 3. Aside from an intermediate increase in lactobacilli, it could not boost the amount of these supposedly healthy bacteria in the long term. Rather than that it did induce an allegedly statistically non-significant decrease in the overall diversity (figure 3, left).

Minor differences with quasi-nonexistent real-world effects

At the mRNA level, the addition of L. planatrum counter-acted the anti-obesity effects of green tea, as evidenced by

Figure 4: Body weight and fat levels of the mice (Axling. 2012)

• 20% higher fatty acid synthase levels, an enzyme that's responsible for the synthesis of fatty acid
• the reversal of the statistically significant reduction in acetyl-CoA caroxylase (ACC), an enzyme that's one step ahead of FAS in the cascade of which you could say that it supplies the raw material for fatty acid synthesis, and
• minimally higher PPAR-gamma levels (responsible for fat storage) 

in the LP + GT vs. GT group, respectively. The net effects on body weight and fat mass, on the other hand were negligible. In essence the bulk of the beneficial effects of the green tea extract remained intact. Moreover, the addition of L. plantarum did have two distinct effects, that were not observed in the GT only group:

Figure 5: Liver cholesterol and HMG-CoA-R after 11 (top) and 12 (bottom) weeks (Axling. 2012)

1. a statistically non-significant -20% reduction in the mRNA expression of the inflammatory marker TNF-alpha, and
2. a whopping and surprising increase in HMG-CoA reductase of +50% and +70% increase in HMG-CoA reductase mRNA compared to the green tee only and the control group, respectively

And while there is nothing in the study that would suggest that there were any beneficial effects from the TNF-alpha reduction, the increase in HMG-CoA reductase is in fact an oddity. After all, despite statistically significant increases in the enzyme that's responsible for the synthesis of cholesterol and the main target of statin drugs (Stancu. 2001), the cholesterol levels dropped by 64% and 39% compared to the control group, in weeks 11 and 22, respectively.

What do these Jerusalem artichokes, agave, bananas, burdock, camas, chicory, coneflower, costus, dandelion, elecampane, garlic,jicama, Leopard's-bane, mugwort, onion, wild yams, yacon and a whole host of other foods have in common? Right! They contain inulin. which has only recently been shown to have the ability to ameliorate body weight gains by up to 50%! Intriguing? Go back to my previous post and learn more about inulin, beta-glucans and their anti-diabesity effects.

Bottom line: A non-statistically significant reduction in TNF-alpha and an elevation of cholesterol synthesis in the presence of lower liver cholesterol levels, which would be suggestive of an increased excretion of cholesterol (thus the increased synthesis to come up for the loss), are in my humble opinion nothing that would render the combination of green tea + L. plantarum superior to the provision of green tea alone. The latter on the other hand, appears to be a great tool to keep the damage of the energy-dense Western diet in check - with no added, let alone synergistic benefit of these particular probiotic.

Maybe the provision of another probiotic or even another strain of L. plantaris would yield at least '1 + 1' results. This would yet be a research question for another study (one I would by the way not be willing to finance ;-) and does not change the fact that the original research hypothesis that there would be a potentiating effect due to the synergism of the two supplements is - even if the scientists don't openly acknowledge that - debunked for L. plantaris DSM 15313 and green tea.

References:

• Axling U, Olsson C, Xu J, Fernandez C, Larsson S, Ström K, Ahrné S, Holm C, Molin G, Berger K. Green tea powder and Lactobacillus plantarum affect gut microbiota, lipid metabolism and inflammation in high-fat fed C57BL/6J mice. Nutr Metab (Lond). 2012 Nov 26;9(1):105.
• Stancu C, Sima A. Statins: mechanism of action and effects. J Cell Mol Med. 2001 Oct-Dec;5(4):378-87.

Green Tea Extracts for Building Strength & Size and Losing Weight - Fact or Fraud? Or, Why It is Always Worth Taking a Look at the Data that Is NOT in the Abstract.

Image 1: If the watery green tea is healthy, then a potent extract must be even more healthy, right? This may well be just another instance of "supplementational idiocy"...

I know people love their Green Tea! After all, Camellia sinensis is one of the staples that has not yet been debunked as another hoax of the supplement of pharmaceutical industry - a real healthfood, right!? Well, you will probably be familiar with my skepticisms towards the notion that taking tons of the polyphenols you are "supposed" to get in relatively small quantities from 2-3 cups of green tea in supplemental form must necessarily be a good thing, just because epidemiological data suggests that people who consume green tea (for those who only now this stuff in capped form: Green tea is actually a beverage ;-) in moderation are overall healthier than people who abstain from drinking hot water extracts (=tea) from minimally oxidized (=green) Camellia sinensis leaves.

Did you miss my previous blogposts on the negative effects of high dose green tea extracts on testosterone levels and male fertility? If so, I suggest you read up on that one before adopting the (stupid) more-is-more principal and popping the whole box of green tea caps at once.

Two recent studies do confirm the notion that green tea, even as a supplement, may be a worthwile addition to your dietary (I suggest you drink the tea not use the supplement) or supplemental (if you cannot stand the taste of the brew) regimen. The first one comes from scientists from the University of Warsaw in Poland (Jówoko. 2011). In a small-scale study with 35 subjects, Ewa Jówoko and her co-workers investigated the effect of 1280mg of green tea polyphenols (from a standardized GTE supplement by Olimp Sports) had on the adaptational response to a standardized 4-week strength training regimen (cf. figure 1)

Figure 1: Summary of the study design - participant characteristics, training and supplementation regimen.

If you look at the summary of the study protocol in figure 1, you may notice that there are two factors which contribute to the real-world significance of the study. Firstly, the subjects followed a semi-standardized diet (90g protein; 270g carbs; 104g fat), because they had to eat at the University's cafeteria. Secondly, the training, as well as the supplementation protocol are similar to what a real beginner would be doing in the gym, when he strives to build lean muscle tissue. On the other hand, this also means that we will probably see different (I would bet even less pronounced) results in trained athletes / advanced strength trainees with an optimized diet and a highly sophisticated supplement regimen - so bare that in mind, when you interpret the following results.

Figure 2: Changes in back squat and bench press 1-RM max and repetition max after 4-weeks of strength training with and without GTE supplement (data calculated based on Jówoko. 2011).

It should not surprise you that 4 weeks of training led to increases in squat and bench press performance. The inter-group differences, as well as the increase in maximal repetition number on the bench, however, did not reach statistical significance. In other words, what we are seeing here are effects of the exercise regimen, independent of GTE supplementation.

Figure 3: Changes in blood pH, base excess and lacate subsequent to the initial (Term I) and post (Term II) muscular endurance and max strength tests (data calculated based on Jówoko. 2011).

Similarly, the changes in blood ph, base excess and lacate subsequent to the initial (Term I) and post (Term II) muscular endurance and max strength tests (cf. figure 3), were not statistically different between groups. But if you followed the SuppVersity news lately, you will already know that blood ph is the domain of plain baking soda... so why even bother with green tea in this regards? After all its not even supposed to be a H+ buffer, but a powerful anti-oxidant, so what we should see are decreased rates of oxidation...

Figure 4: Lipid hydroxyperoxides at rest, 5min and 24h after a strength and endurance test before and after the 4-week intervention (data based on Jówoko. 2011).

And in fact, if you take a very close look at the data in figure 3, you may be able to see (I highlighted the bar for you ;-) why the title of the study, "Green tea extract supplementation gives protection against exercise-induced oxidative damage in healthy men", is not totally off: The degree of lipid peroxidation at rest(!) remained constant (within statistical margin) in the GTE group, while it increased by +27% in the non-supplemented group. In view of the non-existent differences in terms of strength or endurance gains between the groups, it is yet very questionable whether this "protective" effect is worth the 16$ (based on the price of the original supplement used in the study that is only available in Europe) it would cost to mimic the supplementation regimen used in the study - especially for someone who does meet the dietary requirements for vitamin E, which is something the study participants with their cafeteria food didn't.

Scientific fraud in the name of marketing

The results of the second study, which investigated the effects of decaffeinated green tea extract (DGE: 2x530mg per day; 800mg catechins per day, total) on body weight changes in 69 overweight subjects (sedentary males, aged 40–69 years, with BMI > 28 and < 38 kg/m²) are similarly dazzling. In their abstract, A. L. Brown and his collegues from Unilever (do I have to say anything else) summarize the results of their 6-week placebo controlled cross-over study as follows (Brown. 2011):

Despite a similar increase in estimated energy intake during intervention period 1, body weight decreased by 0.64 (SD 2.2) kg and increased by 0.53 (SD 1.9) kg in the DGT and placebo groups, respectively (P< 0.025), suggesting a protective effect of green tea catechins on weight gain.

Does not sound earth shattering, but -0.6kg weight loss does at least appear to be more desirable than +0.53kg weight gain. But even if you do not have access to the full-text (as I do) and are thus able to debunk this as a blatant manipulation of the facts (which, by the way, is the result of selecting data from the more favorable 2 weeks of the 6-week study period), the standard-deviations of 2.2kg in the DGE and the 1.9kg in the placebo group should ring an alarm. If you do have the full-text, and take a closer look at table 3 (cf. excerpt)...

Table 1: Excerpt from table 3 in Brown, 2011

... you should start sensing fraud. After all, the table clearly states that the mean body weight loss over the whole study period was -0.038kg for the placebo and -0.327kg for the decaffeinated green tea group. If you now scroll a few pages down and read the last paragraph...

The authors are all employed by Unilever Research & Development, which is a division of Unilever plc, a company which has a significant commercial interest in tea. Unilever plc provided all funding for the study.

... it should become obvious that, even in the case of something as "innocent" as green tea, you better not believe everything you hear and read about "superfood" and respective extracts on the Internet. So, instead of buying decaffeinated capped bullshit from Unilever, you better go to your local grocery store, get yourself some quality green tea and make tea-time a relaxing part of your probably hectic daily routine ;-)

-20% Reduction in Serum Testosterone by 5 Cups of Green Tea. Endocrine Effects Depend on Catechin Composition.

Image 1: Another SuperFood gone bad? More than 2-3 cups of green tea could do more harm than good to health-conscious male tea-consumers.

While I obviously do not know whether you have come in contact with the rumors surrounding the effects of green tea extract (GTE) on the male reproductive system, I assume that many of you (despite the fact that this is obviously not necessary if you read the SuppVersity news everyday ;-) will be active on one of the various health and fitness related bulletin-boards, where - and here I am certain - the issue of reduced testosterone levels from GTE consumption has certainly been addressed at some point. According to a recent study, published in the September issue Indian Journal of Experimental Biology (Chandra. 2011), your favorite anti-oxidant fat burner Camellia sinensis L., in other words, green tea, has in fact to be counted among those therapeutic agents that despite having been used since ancient times, are not as safe as many customers want to believe.

Figure 1: Molecular structure of the four major green tea catechins (from Kao. 2000)

Did you know that a study from the year 2000 by Kao et al. (Kao. 2000) established a differential effect of green tea catechins (cf. fig. 1) on serum testosterone levels of Sprague Dawley (SD), as well as lean (LZ) and obese Zucker (OZ) rats? While EGCG (and ECG) potently suppressed serum testosterone levels (-69% in SD; -72% in LZ; -69% in OZ), the administration of isolated Epicatechin (EC) and Epigallocatechin (EGC) at a dose of 85 mg/kg BW (human equivalent: 14mg/kg) resulted in a statistically significant increase in testosterone levels of +24% (EC) and +31%, respectively.

In view of the fact that a 2006 analysis of 19 commercially available green tea extracts (Seeram. 2006) revealed that major differences in terms of total catechin content and ratios, the individual effect of "your" green tea caps may be either beneficial or detrimental to your testosterone levels depending on the ratio of (EC+EGC)/(ECG+EGCG) - where a higher amount of the pro-testostosterone fraction, i.e. EC + EGC would obviously be preferable.

According to the results of Amar K. Chandra and his colleagues from Calcutta and West Bengal, green tea belongs to the ranks of plants, such as Neem (Azadirachta indica), Tulsi (Ocimum sanctum) and other agents previously mentioned here at the SuppVersity, all of which have conclusively been shown to hamper reproductive functions and that despite the fact that the neuroprotective, cytotoxic and antioxidant effects of Camellia sinensis L. have been well-established.

Figure 2: Reduction in sperm count after 26 days of consumption of green tea equivalent to 5, 10 or 20 cups of green tea (data calculated based on Chandra. 2011)

As the data in figure 2 goes to show, in the course of 26 days, even the consumption of the equivalent of 5 cups of green tea lead to a statistically significant -2% reduction in sperm count in the gonads of the adult rats who were orally administered with standardized doses green tea after it had been steeped for 15 minutes in 100ml of boiling water, then cooled to room temperature and combined with a second infusion from the same 2.5g of green tea, the catechin composition of which I plotted in figure 3.

Figure 3: Catechin composition and pro- to anti-testosterone ratio of the green tea that was used in the study (Chandra. 2011)

The catechin composition of the specific green tea that was used in the study (the green tea came from the Institute of Himilayan Bioresource Technology) is, as you know from my elaborations on the Kao study in the red "Did you know"-box, probably the determining factor for its effect on the rats testosterone levels. Judged by what I called the pro- to anti-testosterone ratio of 0.47, i.e. the ratio of Epicatechin (EC) + Epigallocatechin (EGC), which increased testosterone in the Kao study, to Epicatechin-3-Gallate (ECG) + Epigallocatechin-3-Gallate (EGCG), which decreased testosterone in the Kao study, the overall effect of the green tea on the testosterone levels of the rats should be negative - and in fact, what Chandra et al. found was a decrease in serum testosterone levels in the "GTE treated groups of animals as compared to their respective control", of which the scientists speculate that it was "due to the impaired synthesis of testosterone."

Figure 4: Reduction in serum testosterone after 26 days of consumption of green tea equivalent to 5, 10 or 20 cups of green tea (data calculated based on Chandra. 2011)

These results also shed a different light on the recently reported "anti-obesity" effect of green tea. After all, the lack of androgens could be an alternative explanation for the GTE-induced 7-20% reduction in body weight Pea et al. observed in their 2011 rodent study and the reduced weight gain (-10% and -14% in the 10 and 20 cup groups, respectively) observed in the study at hand.

Weight loss or lack of weight gain, reduced testosterone, increased LH and all the other negative side effects aside, I would be surprised if low to moderate green tea consumption (2-3cups max.) would actually reduce your chance of conception, hamper your gains in the gym or even induce testicular failure. After all, neither Japan nor China or the other Asian Countries, the inhabitants of which have been having their well-deserved daily cup of (green-)tea for centuries, now, seem to have rampant fertility issues ;-)

Brewing the Perfect Green Tea: Best Brewing Time and Temperature for Optimal Potency

Here is something for all of you who have always asked themselves how to prepare their tea to "get the most out of it". Well, I know normal people would go by taste, but are we normal? No! We want maximal antioxidant-content and a decent amount of caffeine to jack us up and this is where the results from a recent study (Ziaedini. 2010) come into play.

The scientists from the Iranian Institute of Research and Development in Chemical Industries investigated the effect of different brewing times and temperatures on the amount of antioxidants and caffeine you'll get in your tea. Here are the results (cf. table 1):

Table 1: Diffusion coefficients of catechins and caffeine obtained for conventional extraction of green tea leaves.

Obviously it takes a decent temperature of ~70°C to achieve maximal diffusion of the constituents in the short and in the long term. In spite of that, a close look at one of the graphs (cf. figure 1)

Figure 1: EGCG content as a function of time for different water temperatures in °C

shows that even with decent temperatures (>70°C) it takes some time for the antioxidant content (in this case the infamous ECGC content) to achieve significant levels - and lets be honest, do you like, or even, can you really drink a tea that has been brewed for 10 or even 20 minutes? If you can, you're the man (or woman) - if not you probably better buy some capped extract ;-)