Particle Size & LCAT Analysis Shows: Three Whole Eggs per Day Improve Lipid Profile in Men & Women W/ MetSyn. Plus: Up to 700% Increased Lipid Oxidation in Hardboiled vs. Fresh Omega-3 Eggs From Hens on Fish Oil Enriched Diet

"Not the yellow part of the egg!", is the literal translation of "Nicht das Gelbe vom Ei!", which is German and means "not exactly brilliant" - telling isn't it?

As a regular here at the SuppVersity you will probably have come across a line like "make sure to get at least 20g of quality protein (meats, poultry, eggs, fish, dairy) with every meal!" at least once. If we discard the fish, which is, outside of the occasional discussions about mercury and other heavy metals in our food chain, about the only of these highly nourishing foods, the various experts appear to agree is healthy for you, all of them are to be consumed only "on occasion", "in moderation" and preferably in their "lean" or "low-fat" varieties. And while you (should ;-) have read about the beneficial effects of full-fat dairy, yesterday, and about the actually not so bad "bad meats" in the "Meat-O-Logy" post from August 17, 2013, I thought I'd take the forthcoming publication of a paper by Christopher N. Blesso, Jeff Volek et al. as incentive to remind you not to make the mistake and flush the yolk, the best part of the egg, down the toilette.

Recipe for disaster? Take 20 men and women with metabolic syndrome, 5040 whole eggs...

With 40 men and women aged 30–70 years who had been classified with metabolic syndrome the researchers from the University of Connecticut and University of Antioquia in Medellin, Columbia, intentionally picked a representative sample from the ever-increasing number of people with metabolic syndrome for their 12-week dietary intervention trial. I mean, who if not the men and women in this high risk group should suffer from the negative side effects of the cholesterol-laden yellowy, orange-yellow heart killers in disguise?

AHA Definition of Metabolic syndrome

according to Grundy. 2004

Abdominal obesity (waist)
Men	>102 cm
Women	>88 cm
Triglycerides	≥150 mg/dL
HDL cholesterol
Men	40 mg/dL
Women	50 mg/dL
Blood pressure	≥130/≥85
Fasting glucose	≥110 mg/dL

The assumption that subjects with established metabolic syndrome (for the "official" criteria check out the table on the right) is however one of the very few things the study at hand and studies like the one that triggered such an upheaval in August (see Sciencedaily. 2013) have in common. Instead of relying on total, low density and high density lipoprotein levels in the sera of their study participants, Blesso et al. conducted a differential analysis that included paricle number, size (measured by nuclear magnetic resonance spectroscopy), apolipoproteins (apos), oxidized LDL (oxLDL), cholesteryl ester transfer protein (CETP) and lecithin-cholesterol acyltransferase (LCAT) activities at baseline and after 12-weeks on a carbohydrate-restricted diet (25%–30% energy) that contained either  3 regular whole eggs/day (EGG, n = 20) or the volume equivalent in form of a yolk-free egg substitute (manufactured by Sysco Corporation, Houston, TX).

Accordingly, the subjects in the "real egg group" (EGG) consumed approximately 534 mg cholesterol, 0 g carbohydrate, 16 g protein, 12 g fat per day (186 kcal) from hole eggs, while a single serving of the cholesterol and fat-free substitute (SUB) contained approximately 2 g carbohydrate, 14 g protein, no fat, no cholesterol and only 60 kcal.

To minimize possible confounding factors and allow for "blinding" (you can't tell me you don't taste the difference between egg whites and whole eggs, but alas...), the eggs / egg supplements were prepared in advance.

"Compliance was monitored by use of weekly questionnaires and collection of empty product containers. Participants were asked to maintain their normal physical activity, medications, and dietary supplement usage upon starting the 12-week study." (Blesso. 2013)

To be able to track the dietary intake of the subjects, all participants had to fill out 5-day dietary intake records (3 weekdays + 2 weekend days) at baseline, week 6 and week 12 of the study period. Based on the analysis of the respective data, the scientists determined that

• increased relative protein intake from 17.3%± 3.0% to 23.9%± 4.1%
• increased relative fat intake from 38.6%± 6.4% to 45.7%± 7.4%

• reduced total energy intake of -24% for all participants
• reduced relative CHO intake from 40.9 ± 7.4 to 28.3 ± 9.5% of total energy 
• reduced absolute CHO intake from 211.9 ± 51.8 to 114.5 ± 55.0 g/d 

Moreover, there were "significant differences between groups for dietary cholesterol intake (EGG vs. SUB, P < 0.0001) and dietary choline intake (EGG vs. SUB, P < 0.0001)"

• Δ cholesterol intake +106% in EGG vs. -38% in SUB
• Δ choline intake +52% in EGG vs. -12% in SUB

In that, it is important to point out that these changes and their metabolic effect should not be seen in isolation, because choline plays a major role in the metabolism, transport and incorporation of cholesterol into the cell membrane (read more about choline in "Old School Supplements - Choline: Stronger, Faster, Leaner & More Muscular, or Just Another Dumb-and-Barbell Story?").

"So, the men and women in the egg-group did not die?"

Not exactly, no. Rather than being as bad as cigarettes (as the aforementioned "study" that was published roughly 2 months ago would make us believe; Spence. 2013), the consumption of three eggs per day lead to rather favorable changes in the lipoprofile of the EGG consumers:

Figure 1: Lipoprotein serum levels (left), particle size and ratio of LDL to HDL count (right) of the 37 participants who completed the 12-week trial (data expressed relative to baseline; calculated based on Blesso. 2013)

Contrary to the lipoprotein metabolism, which showed differential (and general positive) effects in response to whole egg consumption, the reductions in circulating insulin, HOMA-IR and triglycerides did not show statistical significant inter-group differences:

triglycerides: -29% vs. -18%
• insulin: -21% vs. -14%
• HOMA-IR: -22% vs. -18%

The existing changes were yet on average still somewhat more pronounced in the EGG group, which is an observation you could interpret as joint effect of low carb dieting + egg eating.

The same goes for the increase in LCAT activity the researchers observed only in the EGG group. Since LCAT is critically important in facilitating HDL particle stability and HDL maturation, its elevation in the EGG group

"[...] could be indicative of an enhanced capacity for HDL maturation and may help explain the shift towards larger HDL particles seen with egg feeding" (Blesso. 2013; my emphasis)

and would thus favor enhanced HDL-mediated reverse cholesterol transport from (e.g. the endothelium of your coronary heart arteries) back to the liver.

What about the lipid oxidation?

If we assume that part of the negative effects that have been ascribed to the consumption of egg(-yolks) are brought about by oxidized fatty acids and respective byproducts, it appears wise not to hard-boil your eggs, because the prolonged heat exposure during hard-boiling increases the oxidation of fatty acids. Aside from temperature and duration of the heat exposure, Cortinas et al. also identified the fatty acid composition of the eggs and their vitamin E content as crucial factors that will influence the formation of oxidized lipids. Since the fatty acid composition of the eggs depends on the diet the hens are fed, you would be particularly ill-advised to hard-boil eggs from hens who were fed a diet enriched in fish oil.

As you can see in the figure above the "fish oil eggs" with their readily oxidized long-chain PUFAs had by far the highest  TBA values (expressed as ng MDA/g dry matter) - specifically, if the hens did not receive additional vitamin E that would be transfered to the eggs and reduce the heat-induced oxidation of the long-chain PUFAs in the "omega-3 eggs" during cooking.

So what's the verdict? In conjunction with the aforementioned additive effects on glucose management and the slightly more pronounced decrease in oxidized LDL (both are probably rather a result of the -12% reduction in carbohydrate content of the participants' diets than the effect of the eggs), the statistically significant improvements in LCAT activity in the EGG eater group speak in favor of the notion that three whole eggs a day will still help you keep the doctor away. Whether they will suffice to throw his statins away, as well (see "Eat Whole Eggs All Day and Throw Your Statins Away?"), will yet have to be elucidated and appears overall questionable - and this goes regardless of whether you belong or don't belong to the relatively small subgroup of people who actually benefit from taking a statin, or not.

What stands out of question, however, is that the "side effects" of three egg yolks per day are quite distinct from those of cigarettes, the consumption of which the headline of the aforementioned ScienceDaily.com article implicitly equates with "egg yolk consumption". And just like smoking and eating eggs are two very different pairs of shoes, there is a huge difference between this well-controlled trial, on the one hand, and the undifferentiated observational hokum that got Spence et al. so much media attention back in August, when they concluded "Our findings suggest that regular consumption of egg yolk should be avoided by persons at risk of cardiovascular disease." (Spence. 2013), on the other hand.

References:

• Blesso CN, Andersen CJ, Barona J, Volek JS, Fernandez ML. Whole egg consumption improves lipoprotein profiles and insulin sensitivity to a greater extent than yolk-free egg substitute in individuals with metabolic syndrome. Metabolism. 2013 Sep 26.
• Cortinas L, Galobart J, Barroeta AC, Baucells MD, Grashorn MA. Change in α-tocopherol contents, lipid oxidation and fatty acid profile in eggs enriched with linolenic acid or very long-chain ω3 polyunsaturated fatty acids after different processing methods. J. Sci. Food Agric. 2003; 83: 820–829.
• Grundy SM, Brewer HB Jr, Cleeman JI, Smith SC Jr, Lenfant C; American Heart Association; National Heart, Lung, and Blood Institute. Definition of metabolic syndrome: Report of the National Heart, Lung, and Blood Institute/American Heart Association conference on scientific issues related to definition. Circulation. 2004 Jan 27;109(3):433-8.
• ScienceDaily. Egg yolk consumption almost as bad as smoking when it comes to atherosclerosis, study suggests. August 13, 2013 < http://www.sciencedaily.com­ /releases/2013/08/120813155640.htm > accessed on October 03, 2013.
• Spence JD, Jenkins DJ, Davignon J. Egg yolk consumption and carotid plaque. Atherosclerosis. 2013 Oct;224(2):469-73.

Buffered Alternate Day Fasting + Light Aerobics Cut Body Fat, Maintain Lean Mass & Improve LDL Particle Size. Plus: Conventional Alternate Day Fasting Detrimental for Fertility

This is how the "buffered" alternate day fast works: You eat 1/4 of the White Choc Banana Cream Pie on your fasting day and a whole pie on the next one - well, not really, but the ratios would be right ;-)

I guess at least the SuppVersity facebook friends are probably going to remember the study today's article is dealing with: "Alternate day fasting and endurance exercise combine to reduce body weight and favorably alter plasma lipids in obese humans." To me that sounded like Intermittent Fasting Done Wrong Does Still Produce Great Results, When You Combine It With an Aerobic Exercise Regimen", when I first read the abstract. After taking a brief look at the full-text (thx John!),  however, my perspective on the study changed from "even a dog has its day" to "that oes look interesting, let's see whether this kind of buffered alternate day fasting" is able to take bear up with a classic intermittent fast. 


A brief warning with respect to the data in figure 1: I know that a couple of you are too lazy to actually read the article and do headline + figure hopping. If you intend to to that, please keep in mind that the data in figure 1 is from another recent study, I used to illustrate the negative effects of "real" alternate day fasting.

Not all alternate day fasting is created equal

For the course of this 12-week, randomized, controlled, parallel-arm feeding trial the participants, 61 women and only 3 men aged 25 to 65 years, overweight / obese (BMI 30-39.9kg/m²) and weight stable for at least 3 months were randomized to one of the four study arms (unfortunately the data does not allow for any conclusions, whether the men and women reacted differently to the intervention or "real" alternate day fasting the data from the rodent study I discuss in the interlude surrounding figure 1 shows that this is the case, though):

• control group (C): sedentary, no diet

• exercise group (E): 25, 30, 35, 40min at intensities of 60, 65, 70, 56% of the calculated HRmax (intensity and duration were increased in week 4, 7 and 10)

• alternate day fasting group (ADF): after 4-week controlled feeding period the study concluded with an 8-week self-selected feeding period.
  

  

  As long as study subjects just have to eat what scientists serve them, every diet works. When they are however told to eat a high protein diet, for example, even the reported nutrient intake diverges massively from what the scientists had in mind, when they devised the protocol - you don't believe me? Well, then read for yourself, "What Really Happens, When Science Meets the Real World"

  "During the controlled feeding period (week 1-4) participants consumed 25% of their baseline energy needs on the "fast day” (24 h) and consumed food ad libitum on each "feed day” (24 h). [...] The diet consisted of a 3-day rotating menu plan, and all fast day meals were prepared in the metabolic kitchen of the Human Nutrition Research Unit (HNRU). Fast day meals were consumed between 12.00 pm and 2.00 pm to ensure that each subject was undergoing the same duration of fasting.[...] During the self-selected feeding period (week 8-12) subjects continued with the ADF regimen but no the fast day food was provided to them. Instead, each subject met with a dietician at the beginning of each week to learn how to maintain the ADF regimen on his or her own at home." (Bhuatin. 2013)

  During the counseling sessions the participants were also instructed to make (I quote) "healthy foodchoices on the ad libitum feed days by choosing low fat meat and dairy options and
  increasing fruit and vegetable intake." (Bhutani. 2013)

• combination group (ADF + E): combination of alternate day fasting and exercise protocol

Table 1: Nutrient composition of the fast day diet provided to the combination and ADF groups (Bhutani. 2013)

So, as you've learned from the above overview this was no "alternate day fast", as I had expected it to be, when I read the term "Alternate Day Fasting (ADF)" in the abstract of the study. With 450kcal/day and a nutrient composition providing 50% of the energy in form of carbohydrates, and 25% (each) from fat and protein the "fasting days" were characterized by a low caloric intake (see table 1), but should not have triggered a similar starvation response as the classic alternate day fasting regimen in a recently published rodent study by Kamur et al. from which the data in figure 1 is derived.

Let's briefly take a look why "real" alternate day fasting may not be a good idea

As you can see the effect of the alternate day fasting regimen in the Kumar study, in the course of which the rodents simply did not receive any food for a whole day, had profound negative effects on the hypothalamo-gypophysial-gonadal axis - especially in the female rodents.

Figure 1: Effects of every-other day fasting vs.ad libitum feeding on ovarian weight, estradiol, lutenizing hormone, leptin, and testosterone in female and male rodents (Kumar. 2013)

Now the first thing that will jump your eye is certainly the profound increase in serum estrogen in the IF group. Looks strange and certainly not anywhere near what you would have expected right? If you think that this cannot be that bad, you are probably a man, is that right? I see... so what really is bad about the estrogen explosion is actually that it is "living proof" of the total disruption of the cyclic interplay between estrogen and progesterone that's at the heart of female (in-)fertility (on a side note: the male rodents simply got skinny fat on that regimen).

Now why didn't these adverse events occur in the study at hand?

Actually I am only assuming that there were not similar negative effects in the study at hand. After all, we don't have the respective hormonal and even in the rodents it took it's time (2 cycles) until the females were totally infertile. Changes like these would thus probably have gone unrecognized. I do however suspect that the major factors contributing to the overall hormonal decline in the rodent study were the extendended time-span without food and the overall caloric deficit, which is usually ~30-40% of the habitual intake in rodents on alternate day fasting regimen, as they do not compensate for the fasting day on the subsequent day.

Figure 2: Changes in body composition in the combined, ADF, exercise and control groups (Bhutani. 2013)

With the extended fast being absent and a ~75% reduction in calorie intake on the fasting days, only (humans are unfortunately much better in "compensating" for a lack of food on a fasting day, anyway ;-) and the huge amount of adipose tissue the rodents in the Kumar study obviously did not have as an "emergency reserve", it is therefore unlikely that the beneficial changes in body composition Bhutani and his colleagues observed in their 64 subjects (see figure 2) were accompanied by hormonal deteriorations (although 61 of them were, as previously mentioned, female).

Figure 3: Changes in lipid profile (left) and glucose metabolism in the combined, ADF, exercise and control groups (Bhutani. 2013)

It is nevertheless interesting to see that there were no significant improvements in fasting glucose, insulin or HOMA-IR (the measure of long-term blood glucose levels; not shown in figure 3) in any of the groups. Moreover, the improvements in total LDL and HDL cholesterol were only significant in the combined group and that the total amount of the inflammatory marker CRP remained essentially the same in all four arms of the 12-week intervention study.

Image 2: Ramadan fasting can serve as a relatively well studied "model" of intermittent fasting. With the additional restriction of water intake and the common practice of rising early to have breakfast there are yet non-negligible differences. You can find more information about the strengths and limitations of this model in Part 2 and Part 3 of the IF series.

A brief reminder for everyone who missed the Intermittent Thoughts on Intermittent fasting series back in the day: If there is one thing we can take away from studies investigating the effects Ramadan fasting (a Muslim fasting ritual, where you eat only when the sun goes down) it is that eating the same amount of food at different times of the day alone is not going to make you lose weight. If you do intermittent fasting in order to lose weight you still have to achieve a caloric deficit. Plus, when you are doing it during a bulk, I personally suspect that you are more likely to gain body fat, simply because your body cannot make "good use" (=muscle glycogen and skeletal muscle protein) from all the food you will be cramming down in a small fasting window. So, if you insist on IF on a bulk, do at least increase your feeding window to 8h to be able to spread your energy intake more evenly.

Based on a comparison of the data from all three groups, we can yet also conclude that it is the alternate day fasting (or probably rather the caloric deficit) that exerts the beneficial effects on what the scientists subsume under the umbrella term "CHD risk indicators":

"the combinationof ADF plus exercise decreased LDL cholesterol (12% from baseline) while increasing HDL cholesterol (18% from baseline); a change that was not noted for any other intervention. The combination group also experienced an increase in LDL particle size, and a  reduction in the proportion of small LDL and HDL particles." (Bhutani. 2013)

Only later in the discussion of their results do they mention that those "CHD risk indicators" improved to the same extend in the ADF only group - with even more significant changes in the particle profile.

---

Learn why breakfast is probably not the "most", but rather the least "important meal of the day" (read more).

Bottom line: The weight and fat loss are certainly intriguing. The regimen appears to be reasonably easy to follow and the fact that the improvements in glucose metabolism were marginal could well be related to the fact that the subjects were - despite being seriously overweight (!) - not diabetic.

Keeping all that in mind I am still not convinced that a regular intermittent fasting routine (16-18h fasting window on every day; learn more about intermittent fasting here at the SuppVersity) and a combined aerobic + strength training regimen would not have yielded even superior results. The average physical culturist probably will be better of with this by now almost "classic" diet routine.

References:

• Bhutani S, Klempel MC, Kroeger CM, Trepanowski JF, Varady KA. Alternate day fasting and endurance exercise combine to reduce body weight and favorably alter plasma lipids in obese humans. Obesity (Silver Spring). 2013 Feb 14.
• Kumar S, Kaur G. Intermittent Fasting Dietary Restriction Regimen Negatively Influences Reproduction in Young Rats: A Study of Hypothalamo-Hypophysial-Gonadal Axis. PLoS ONE. 2013; 8(1): e52416.