The Oiling of the Liver: The Good & Bad Short- & Long-Term Effects of Tocotrienol + Carotenoid Laden Red Palm Olein, Regular Palm-, Corn- and Refined Coconut Oil

I would not expect "red palm olein wonders", but more RPO and less corn oil in the American diet may at least buffer the liver disease burden in the US (the figure is based on data provided by the American Liver Foundation)

On Turesday, November 19, 2013, you've learned from a study by Subermaniam et al. about the "anti-rust" effects of coconut oil (if you missed that, you can catch up here), today, we are going back to Malaysia and the Universiti Kebangsaan Malaysia and the results of another team of researchers to learn about the effects the various oils have on the "oiling of the liver" (Dauqan. 2013).

I guess most of you will remember my previous comments about the critical role of the liver (and its health or disease) in the development of the metabolic syndrome (read it up). It is thus by no means irrelevant, whether the chronic ingestion of a certain type of oil will result in MDA levels of 92µmol/g or  27.3µmol/g.

Boring!? No, rather surprising!

If you think this sounds boring and are by no means surprised that the malondialdehyde levels of the liver samples the researchers harvested after 4 weeks were 27.3µmol/g, 92µmol/g, 54µmol/g, 47.4µmol/g and 72.6µmol/g for the control diet with mixed fats, red palm oloein (RPO), regular palm oil (PO), corn oil (CO) and the previously celebrated coconut oil (COC), respectively, I would suggest you have a closer look at the the "magic" that happened over the following 4 weeks of on 15% RPO, PO, CO and COC diets.

Figure 1: MDA levels (µmol/g) of liver tissue as a marker of lipid oxidation after four and eight weeks on control diet or control diet with 15% of red palm olein, palm oil, corn oil or coconout oil (Dauqan. 2013)

Well, you see, the way the effects of red palm olein came full circle after another months on the 15% RPO diet is hardly "boring", is it? The MDA levels, a relatively reliable indicator of local lipid oxidation, of the rodents on the 15% red palm olein diet is now, 4 weeks after peaking at 92µmol/g down to 25.2µmol/g, indicating that the level of lipid peroxidation in the livers of the RPO group is now significantly lower than that of any other group (43.4µmol/g, 50.1µmol/g and 48.3µmol/g for control, palm oil, corn oil and coconut oil).

Short term detriments, long term benefits!

I know it sounds more than awkward, but eventually every SuppVersity student should be aware of the fact that the extrapolation of long-term effects from short-term data is a 'risky' business. Unfortunately, even 'experts' often disregard this fundamental rule, when they formulate their recommendations on nutrition, supplementation and exercise.

Table 1: Carotenoid  and vitamin E composition (in %) of crude palm oil and red palm olein; the data is from a different study by Bonni & Choo who tested commercially available products (Bonni. 2000)

The statement, "Prefer coconut oil and avoid red palm olein!", for example would have been a reasonable dietary if we did not know about the turn-around in the second part of the study, when the beneficial effects of the saturated fat content of the coconut oil begin to fade and the absence of natural anti-oxidants in refined coconut oil begins to show its ugly face. At this point, the moderate amount of unsaturated fats in red palm olein (13% omega-6, 0% omega-3; see Bonnie. 2000), of which I am honestly not sure if it is the actual reason of the initial increase in lipid peroxidation (remember: corn oil has more PUFAs!), or whatever other underlying cause of the initial rise in inflammation is overriden by the accumulating amounts of vitamins E and carotenoids from the red palm olein, which rendered the liver of the oxidation-proof, or "rustless" if you will - similarly rustless as the hearts of the rogents in the previously cited study by Subermaniam et al. (learn more).

200g of palm fruits have the same amount of tocotrienols as 4kg of oats. Learn more "tocotrienol" and red palm oil facts in "Tocotrienols: What They Are, What They Do & How They Work + Why the RDA of Palm Olein is NOT 1xCup Per Day " | more

Bottom line: I would like to formulate two take home messages for today's SuppVersity article. Firstly, a theoretical one, which shall remind you of the fact that you can do more harm than good, if you (accidentally) terminate a study in a transitional state and formulate long-term dietary recommendations based on short-term observations, because the study at hand clearly indicates that some effects - in this case the antioxidant effects of the tocopherols, -trienols and carotenoids - take their time to become measurable. And seconfly a very practical one, which is eventually only a reminder of the existence of red palm oil (see article referenced on the right) - an excellent source of dietary antioxidants and probably your only chance to get your tocotrienols and high(er) amounts of some of the rarer carotenoids from regular foods.

References: 

• Bonnie, T. Y. P., & Choo, Y. M. (2000). Valuable minor constituents of commercial red palm olein: carotenoids, vitamin E, ubiquinones and sterols. Journal of Oil Palm Research, 12(1), 14-24.
• Dauqan, E., Abdullah, A., & Sani, H. A. (2013). LIPID PEROXIDATION IN RAT LIVER USING DIFFERENT VEGETABLE OILS. Malaysian Journal of Analytical Sciences, 17(1), 300-309.
• Valls, V., Goicoechea, M., Muniz, P., Saez, G. T., & Cabo, J. R. (2003). Effect of corn oil and vitamin E on the oxidative status of adipose tissues and liver in rat. Food Chemistry, 81(2), 281-286.

Standard American Diet Has 'Optimal' Fatty Acid Ratio to Induce Diabesity. Plus: Study Shows Doubling Saturated Fats Would Yield More Benefits Than Halving Them

Study confirms: The SAD diet yields 'optimal' results (img. forbes.com)

Since this post is already lengthy enough, I will spare you how saturated fatty acids have long falsely been accused as the sole driving force of the western obesity epidemic and how the tides appear to be slowly yet steadily appear to be turning, as scientists delve deeper and deeper into the interactions of the total fat content in the diet, its fatty acid composition and the interaction of both with the two other macronutrients and their specific forms and get right to the study at hand. A study that appears in the current issue of the Journal of Lipid Science and deals with the first of the aforementioned interactions. The one that focuses on the total fat content and the individual fatty acid make-up of the diet (Enos. 2012).

Fat shoot out: Saturated vs. mono vs. PUFA

As Enos et al. point out, the main purpose of their study was to examine the effects of three high fat diets differing only with respect to the percentage of total calories from saturated fats.

• SFA-6% - contained 6% saturated fats,
• SFA-12% - contained 12% saturated fats, and
• SFA-24% - contained 24% of saturated fats

While the the high fat diets were set to have an identical fat (40% of the energy), carbohydrate (45% of the energy) and protein content, the two control diets were low in total fat (12%/68%/20% of the energy from fat/carbs/protein). They did however likewise differ as far as their fatty acid composition is concerned, with the modified chow mirroring the ratios (!) not the amounts of mono- and polyunsaturated fatty acids of the high fat chow (see figure 1).

Figure 1: Fatty acid composition (left) and their sources (right) that were used in the different diets the rodents were fed for 16 weeks (based on Enos. 2012)

The diets were administered for 16 weeks. Body composition and metabolism (glucose, insulin, triglycerides, LDL-C, HDL-C, total cholesterol) were examined monthly.  Adipose tissue (AT) expression of marker genes for M1 and M2 macrophages and inflammatory mediators (TLR-2, TLR-4, MCP-1, TNF-α, IL-6, IL-10, SOCS1, IFN-γ) was measured and so on and so forth... and the results were... well, not exactly as you may have expected (the latter statement assumes that you expected the SFA to be either the savior or the doom of the human race, depending on which side of the LC/LF divide you are stading).

Figure 2: Body composition (left), adipocyte size (right) and fat pad weight (inset) of the rodents at the end of the study period (Enos. 2012) Values not sharing a common letter (abc) differ significantly over time within the given diet treatment (P≤.05)

If you take closer look at the data in figure 2, there are two things that will probably catch your eye right away. The first 'eye catcher' pertains to the influence of replacing a large amount of the omega-6 fatty acids by monounsaturared fatty acids, as you will find them in olive oil, for example.

• The rodents who received the modified standard chow, with a fatty acid composition identical to the high fat diets (SFA-6%, SFA-12%, SFA-24%) had the exact same body composition as their mates who received the standard chow with its 3.7x higher n6:n3 ratio. The removal of omega-6 fatty did thus not have any beneficial effects on adiposity in the low fat groups.

The second 'eye catcher' is the non-linear increase in adiposity with increasing amounts of saturated fatty acids in the diets. This does not mean that the expected increase in obesity and adipocyte size was totally absent (read the latest "Get Lean & Stay Lean" item for more information about the association of large fat cells and metabolic syndrome), though:

• The mice in the SF-6-24% did all gain significantly more body weight and body fat than their peers on the low fat diets, but there appears to be a turning point, when the saturated fat content exceeds 12%. After all the mice in the SFA-24% group had almost the same body composition as their peers on the SFA-6% diet.

So, what do we make of these 'eye catchers'? The first one, you could argue, shows that "omega 6 overload" is not a problem, as long as you are consuming a low fat diet, in the first place. Even with the major part of those 12.2% of energy your diet provides in form of various fatty acids belonging to the potentially inflammatory omega-6 fatty acids, that's still way too low to do any harm. It does, by the way, yet explain why low fat diets work so well in a society, where most high fat foods the public consumes are laden with omega-6 fatty acids - not an insignificant result, I would say.

The 12%-SF diet, most closely mimics the standard American diet

Apropos public, the second 'eye catcher' is even more telling in term of public health,... wait, I should write sickness. Why? Well, the 12%SFA high fat diet, which supplies ...

• 47% of energy in form of carbohydrates (380g sucrose, 100g maltodextrin, 50g cornstarch per 1kg of diet; identical for all SFA groups),
• 40% of energy in form of fats (of which 12% were saturated fats), and
• 13% of energy in form of protein (from casein),

... mimics, as the researchers point out, "most closely" (Enos. 2012) the standard American diet (SAD). And the result is obvious: Diabesity!

It's a fat balancing act of macro and micro ratios  - complex and far from being understood 

What's intriguing though, is that the adipogenic effects of the diet were ameliorated, when the SFA content was further increased and the diet contained 68.6g of lard per kg chow instead of just 35.4g and 96.7g of coconut oil instead of just 30g. Since this increase in SFA was at the expense of both mono- and omega-6 fatty acids, you could of course also argue that replacing at least the latter of the two with SFAs must be healthy. Unfortunately, even a brief glance back at figure 2 reveals that this is not necessarily correct. After all, the SFA-6% group was still better off than the SFA-24% group, although they had the highest amounts of oleic and omega-6 fatty acids in the diet.

By now you should actually have realized that this is once more a difficult balancing act. Where different baseline intakes of dietary fat and carbohydrates (total) are pair of setscrews and the individiual fatty acid composition of the diet is another one. And the way these setscrews are set will not just influence the body composition:

Figure 3: Serum IL-6, MCP-1, adiponectin and leptin levels, TNF-alpha mRNA expression in the adipose tissue (left), adipose tissue sample form the rodents receiving standard chow, the SFA-12% and the SFA-24% diet (Enos. 2012). The fat cells of the SFA-6% animals looked similar to those on the SFA-6% diets.

Based on the body composition data presented in figure 2 the marked increases in serum leptin and TNF-alpha mRNA expression in the adipose tissue of the rodents in figure 3 (left) should be about as unsurprising as the fact that the adipocytes of the SFA-12% group show the greatest macrophage infiltration and subsequent necrotic tissue.

If anything is surprising, it is the non-significance of the peak in IL-6 in the SFA-24% group (this was due to a very high standard deviation) and the fact that the serum level of MCP-1 a marker of increased macrophage activity was not elevated, while the adipose tissue mRNA expression was significantly higher (5-8x) in all SFA groups compared to both of the control diets. In the end this is yet only another clear sign that far more processes than we have previously thought happen locally and do not depend on circulating and thus endocrine signaling molecules.

Figure 4: Blood glucose and insulin levels of the mice over the course of the study period (Enos. 2012)

If you take the data from figure 4 into account as well, you will certainly agree with the statement Enos. et al. make pertaining to the negative effects of the SFA-12% diet, which is - just to remind you - the mirror image of the standard American diet:

"The 12%-SF diet, most closely mimicking the standard American diet, led to the greatest adiposity (absolute fat mass), macrophage infiltration, and IR [insulin resistance]." (Enos. 2012)

Figure 5: Total  cholesterol (TC, top) and LDL-C to HDL-C (bottom) ratios (Enos. 2012)

And I guess it would actually be about time to get to the bottom line, here, if it was not for the sentence that follows this assertion:

"Although the 24%-SF diet increased adiposity and produced IR, it did not significantly increase macrophage infiltration, it led to a lesser degree of AT inflammation, and it did not raise the TC/HDL-C ratio." (Enos. 2012)

Yep, you are reading right, as the data in figure 5 shows the total to HDL ratio of the SFA-24% group, which were those rodents who consumed the largest amount of "bad" saturated fat, was virtually identical to the one of the rodents on the standard and the modified standard chow and significantly lower than in those rodents who 'lived the American way of life' (SFA-12%). A similar trend was seen in the LDL:HDL radio and the triglyceride levels.

Bottom line: So, does that mean that we would just have to fry our potato chips in lard and all will be good? Not really, no. If we keep munching tons of plain sugar, even a saturated fat only diet is not going to save us from doom (I suspect there will be another inflection point at levels which exceed 50% SFA, anyway). What the study results do yet clearly implicate is that the macronutritent and fatty acid composition of the standard American diet is downright conspicuously obesogenic, pro-diabetic, inflammatory.

While the macronutrient ratio (high carb + high fat) appears to set the body into fat storage mode, the individual ratios of the fatty acids determine the efficacy of body fat storage, the negative effects on blood glucose management, and the degree of adipose tissue inflammation - and the standard American diet excels in all these disciplines.

As far as the saturated fats go (I wonder if it also plays a role that one of the main sources was coconut oil), the study suggests that you can achieve ameliorations of adiposity on both sides of the 'obesogenic optimum' of 12% saturated fats. If you take a last look at the data in figure 4, you will yet have to concede (or triumph?) that eating more not less saturated fat and thus frying your potatoes in lard, appears to be the more promising modification you could make, if the saturated fat content of the diet was your only set screw. Feels good to know it isn't right?

References:

• Enos RT, Davis JM, Velazquez KT, McClellan JL, Day SD, Carnevale KA, Murphy EA. Influence of Dietary Saturated Fat Content on Adiposity, Macrophage Behavior, Inflammation, and Metabolism: Composition Matters. J Lipid Res. 2012 Oct 28.

+87% Increase in Testosterone Within 21 Days from a 100% Natural Supplement? Study Shows: Soy Bean Extract Can Do Just That While Wreaking Havoc on Your Testes. Plus: Corn Oil Reduces Testosterone to Estrogen Ratio by -50%!

Image 1: I guess the feed of those boars does not contain any corn oil and is spiked with both bisphenol A and soy bean extract - I mean, how else could you possibly explain those balls? (img dirtybutton.com)

Let me start today's post with a few questions: Would you buy a 100% natural product that can lower your estrogen levels by up to -98%, increase the weight of your testes by ~30% and, above all, boost your testosterone levels by a whopping +87%? I guess, at least all those of you who either have not read or not understood the Intermittent Thoughts episode on estrogen's role in skeletal muscle hypertrophy are just sitting there, nodding their heads... I would yet also venture the guess that this nodding will end pretty abruptly, now that I am about to tell you that this all natural testosterone booster is derived from the powder of 2kg of Glycine max soy beans via methanol extraction, subsequently freeze dried and capped into 600mg caps of which the average adult (~80kg body weight) is supposed to ingest two per day.

Chose your poison: BPA, soy, or maybe just some governmentally subsidized corn oil?

The preceding paragraph was an ironic, yet as far as the underlying facts and figures are concerned 100% accurate introduction to today's post which revolves around a study Evanski from the Mind&Muscle forum has brought to my attention (Norazit. 2012). The authors, a group of scientists from the University of Malaya in Kuala Lumpur, Malaysia, had set out to investigate the purportedly negative effects of what they call "soya bean extract" (interestingly this spelling of "soy", which is identical to the German version is probably the reason the study did not appear on my "interesting stuff for the SuppVersity radar", before ;-), bisphenol A, 17β-estradiol and "harmless" corn oil on the testis and endocrine system of juvenile rats.

Figure 1: Phytoestrogen content (µg/g dry weight; mind the logarithmic scale!) of soy bean extract an standard rat chow measured by LCMS (data adapted from Norazit. 2012)

To this ends, the scientists divided thirty 21-day old juvenile male Sprague-dawley rats (=the standard lab rat) into five groups, receiving either a standard diet (which contained an insignificant amount of soy, cf. figure 1) + 100mg/kg Tween 80 (a standard food emulsifier with derived from polyethoxylated sorbitan and oleic acid; this group served as control for the soy and the bisphenol group) or standard diet +100mg/kg of corn oil (Mazola; this group served as a control for the estradiol group because the 17b-e did not dissolve in the Tween 80), soy extract, bisphenol A (Aldrich Chemical Co.) and 17b-estradiol (the most active form of estrogen, which binds to both the alpha- and beta-receptor) for three weeks.

Note: It is (at least in my view) a lucky coincidence that contrary to the soy extract and the bisphenol, the estradiol did not solve in the Tween 80, so that the scientists had to come up with Mazola corn oil as a "positive control". I mean, if you take a look at the effects this supposedly neutral "solvent" had on the endocrine milieu of the peripubertal rats, it is no wonder that with the average testosterone levels of the male inhabitants of the #1 corn producer of the world, the Unites States of America, is on a constant decline.

At the end of the study period the rats were sacrificed, the testis were excised and their testosterone and estrogen levels were assessed using standardized enzyme immunoessay (EIA) kits from Caymen Chemical.

Figure 2: Section of seminiferous tubules from control Tween 80 group, BPA group and soy bean extract group; (1) maturing spermatids, (2) lumen filled with cellular debris, (3) vacuaolation, (4) interruption of spermatogenesis (data adapted from Norazit. 2012)

Even a layman can see that both the bisphenol A, as well as the soy bean extract treatments induced profound changes in the cell-morphology of the testes (cf. figure 2). Vacualation (3), i.e. formation of vacuoles in cellular tissue, was present in both, only the bisphenol A group showed the characteristic lumen filled with cellular debris (2). Visible signs of spermatogenesis (1) were visible in neither of the groups, a clear interruption of the latter (4), was yet observed only in the soy and the estrogen group (latter not shown in figure 2). Moreover, the estrogen treated animals were the only ones where the testis showed clear signs of apoptosis (cell death).

The "harmless" corn oil shifts the testosterone to estrogen ratio from ~1/1 to 1/2pg/ng

Reckless, as I am I decided to discard Norazit et al.'s distinction into the BPA and soy groups with the Tween 80 group as a control and the estradiol group with their corn oil control and just plotted the total body and total and relative testis weight gain, estrogen and testosterone levels relative to the Tween 80 group. In other words, I treated the corn oil group as if it was just another treatment group. This is obviously somewhat fishy, but if no scientist appears to be willing to investigate the potential negative effects of corn oil on the endocrine system of adolescent rodents (let alone humans), this is the only way for us to get respective data ;-)

Figure 3: Body weight gain, total and relative right testis weight, estradiol and testosterone levels in peri-pubertal rats after 21 days on diets containing 100mg/kg bisphenol A, soy bean extract, corn oil or 17b-estradiol (in soy bean oil); data expressed relative to Tweenn 80 (polysorbate + oleic acid) control (data calculated based on Norazit. 2012)

And if you take a look at the data in figure 3 (the vertical axis of which is by the way discontinuous!) it becomes clear that you better feed your boys a food solvent such as Polysorbate 80 (=Tween 80) than the "healthy" corn oil the US government is trying to con you into. After all, the administration of 100mg/kg corn oil (human equivalent ~16mg/kg) during puberty decreased the testis weight of the rats by -23% it reduced the amount of estradiol by -35% and the amount of testosterone by -66% and thusly shifted the testosterone / estrogen ratio in the peri-pubertal rodents from 1.11pg/ng to 0.57pg/ng!

BPA and soy compete for the title of "most potent endocrine disruptor"

Following the bro-scientific "the more the better" type of reasoning, bisphenol A and soy bean extract are two potential candidates for the "testosterone booster of the year"-award. After all both, the organic solvent bisphenol A, as well as the "natural toxin" (sorry, I just had to write that ;-) soy, exert potent (8x) and ueber-potent (100x) effects on the testosterone to estrogen ratio, which is 8.2pg/ng for BPA and 100.1pg/ng for soy!

Note: Neither I, nor the scientists have any clue as to why the results of this study are diametrically opposed to those of previous studies in which extracts from soy products reduced, not increased, testosterone levels in male rodents and monkeys(!), across-the-board (eg. Sharpe. 2002; Cline. 2004) - and that although Sharpe et al. observed an increase in the testosterone producing Leydig cells in their soy-formula fed monkeys. Whether the rats in the study at hand were in a state where similar effects temporarily increase testosterone output until the Leydig cells literally "burn out", or whether other effects were responsible for the temporary increase in testosterone, would have to be elucidated in future studies, the results of which you will obviously read here at the SuppVersity, first ;-)

So, even if we assume that the data is correct and there were no cross-reactions between components in the soy bean extract and the testosterone anti-body test, I would strongly caution against the use of either of this compounds to boost your testosterone levels - I mean what's the use of a wickedly skewed testosterone to estrogen ratio (which in and out of itself will probably mess up your health and can potentially hinder your gains, cf. "Are You Serming Away Your Gains?"), when, at the same time, your testicles turn into dysfunctional balloons?

Body Fat Modulation with Corn Oil & L-Carnitine: What You Can Learn From Your Schnitzel

Its quite remarkable that, primates aside, swine are among the best models of human metabolism. So, even if you do not feel piggy at all, the fact that pigs just as humans are omnivores, makes them a much better model for metabolic disease than rodents. It is thus not too unrealistic to assume that we can learn something about ourselves from the results of a very recent study published in the Journal of Animal Science (Apple. 2011).

Figure 1: American Pork Cuts; quality is determined by corn-oil and carnitine intake of the swine.
What lessons can you learn from our pink relatives?

Investigating the effects of l-carnitine supplementation on the quality characteristics of fresh pork bellies from pigs fed three levels of corn oil, J.K. Apple and his co-workers observed a linear trend towards decreased belly-firmness with increasing amounts of corn oil (0, 2 or 4%) in the diet. If you look at the average American, his/her high corn oil consumption and their respective (pot-)bellies, this should not surprise you. All aesthetic considerations aside, those feisty pot-bellies are nothing but the outward sign of metabolic derangements that - without appropriate lifestyle interventions - have their owners suffer from diabetes, high blood pressure, chronic inflammation, and all the other players in the (eventually) deadly "game" of metabolic syndrome.

[...] belly firmness decreased linearly (P < 0.001) with increasing dietary OIL, but there was no (P ≥ 0.137) effect of CARN on any belly firmness measure.

Now, did the touted fat-burner l-carnitine prevent these effects? No, it didn't. Yet, what it did do is it increased the amount of saturated (SFA) and mono-unsaturated (MUFA) fatty acids and decreased the amount of polyunsaturated fatty acids (PUFA) in the belly tissue:

Dietary CARN increased (P < 0.05) the proportion of total SFA in the intermuscular fat layer, increased (P < 0.05) the proportion of total MUFA in the primary and secondary lean layers, and decreased (P < 0.05) the proportion of total PUFA in the intermuscular fat and secondary lean layers of pork bellies.

In view of the finding that increasing the amount of corn oil in the diet tended to increase the PUFA content of the belly tissue, while depositing the highly oxidative polyunsaturated fatty acids preferentially in fat and not lean layers, one must acknowledge that L-carnitine, despite not being able to prevent the outwardly visible (and touchable) negative effects of a diet high in omega-6 rich corn oil, was yet able to modulate the effects of excess PUFAs on intra-tissue body fat composition.

Against the background of the recent changes in the scientifically accepted perspective on the previously vilified saturated fatty acids and possible beneficial effects on cell stability and inflammation the significance of these results goes beyond profane insights into the management of pork quality and solidify the foundation of my previous recommendation to avoid omega-6 instead of increasing the overall PUFA load by additional omega-3 supplementation. What's new, however, is the role l-carnitine supplementation may play in your efforts to get rid of overly high tissue levels of omega-6, since the reduced storage in fat tissue and the increased storage in muscle could be able to (a) decrease inflammation of the fat tissue and, at the same time, (b) increase oxidation of PUFAs in exercised muscle tissue. Yet, without appropriate dietary changes and the incorporation of regular exercise sessions into your  new, healthier lifestyle all carnitine in the world won't help you, if you insist on eating too many breaded and fried schnitzel with French fries and a boatload of mayonnaise and ketchup.