Author Archives: Bill

Fasting, circadian biology, and epigenetics

From the best I can gather, one of the more immediate players in circadian biology is the coenzyme nicotinamide adenine dinucleotide (NAD), which participates in a variety of redox reactions.  Fasting increases the intracellular NAD/NADH ratio, setting off a cascade of events involving epigenetics and the regulation of metabolism.

NAD activates sirtuins, a family of deacetylase enzymes.  This is epigenetics.

SIRT1

 

SIRT1 regulates the activity of BMAL1 and CLOCK, two circadian transcription factors, which target NAMPT, an enzyme that synthesizes NAD.  And in a curious feed-forward mechanism, CLOCK and BMAL1 enhance SIRT1 expression… genetic deletion of any of these players induces insulin resistance (Zhou et al., 2014), and this can be recapitulated with constant darkness: reduced BMAL1 and SIRT1, hepatic insulin resistance; the latter can be reversed with resveratrol (which may or may not be acting through SIRT1; this is controversial).  While alcohol does no great favors for circadian biology, if you’re going to imbibe, perhaps a resveratrol-rich Argentinian malbec served, and this might be the important part, at night, when all of this stuff is going on… coincidentally [fortunately], that’s precisely when most choose to imbibe.

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More on physical performance and ketoadaptation

The various studies on how low carbohydrate diets impact physical performance are very nuanced.  Here’s what I mean by that.

Exhibit A. Phinney 1980

Phinney 1980

In this [pioneering] study, obese patients were subjected to a variety of performance assessments in a baseline period, then after 1 and 6 weeks of weight loss via protein-sparing modified fast (1.2 g/kg ideal body weight from lean meat, fish, or fowl; probably around 80 grams of protein/d, 500-750 kcal/d). They lost a lot of weight, 23 pounds on average, two-thirds of which was body fat. There was no exercise intervention, just the performance assessments.

During the ‘exercise to exhaustion’ treadmill exercise, RQ steadily declined from baseline to week 1 to week 6, indicating progressively more reliance on fat oxidation.  This was confirmed via muscle glycogen levels pre- and post-exercise: during the baseline testing, they declined by 15%; after 6 weeks of ketoadaptation, however, they only declined by 2%, while ‘time to exhaustion’ increased by 55%.  After only 1 week of the diet, time to exhaustion plummeted, as expected, by 20%.

This was, as mentioned above, a pioneering study in the field of ketoadaptation. It also challenges one of the prevailing theories of ‘fatigue’ …while carb-adapted, the subjects fatigued after 168 minutes, with muscle glycogen levels of 1.29 (reduced by 15%); while ketoadapted, they fatigued after 249 minutes with muscle glycogen levels of 1.02 (reduced by 2%).  In other words, they had less glycogen to begin with, used less glycogen during exercise, and performed significantly better (running on fat & ketones).

Exhibit B. Vogt 2003

Highly trained endurance athletes followed a high fat (53% fat, 32% carbs) or high carb (17% fat, 68% carbs) diet for 5 weeks in a randomized crossover study. In contrast to Phinney’s study, these participants were: 1) highly trained; and 2) exercised throughout the study.

Maximal power output and VO2max during a similar ‘time to exhaustion’ test was similar after both diet periods.  Same for total work output during a 20 minute ‘all-out’ cycling time trial and half-marathon running time.  Muscle glycogen was modestly, albeit statistically non-significantly lower after ketoadaption; however, ketoadapted athletes relied on a higher proportion of fat oxidation to fuel performance as indicated by lower RQ at every level of exercise intensity:

Vogt RQ

Again, this is the essence of ketoadaptation. Physical performance as good as or better using fat and fat-derived fuels.

One reason Phinney’s glycogen-depeleted ketoadapted subjects may have done so well is their reliance on ketones (probable) and intramyocellular lipids (IMCL) (possible).  In Vogt’s study, IMCL increased from 0.69 to 1.54% after ketoadaptation…

Also, food intake and body fat declined, and training volume increased in the low fat group; whereas food intake increased, and body fat and training volume declined in the high fat group.  Reminiscent of anything?

High fat, low carb -> eat more, exercise less, STILL LOSE BODY FAT.

Vogt data

Sorcery?  No.  Diet impacts more than just mood and body composition – resting energy expenditure increased in the ketogenic dieters.  This isn’t an isolated finding.

Exhibit C. Fleming 2003 

This was another study in non-trained athletes, consuming high fat (61% fat) or control (25% fat) diets for 6 weeks.  The tests were the 30-second Wingate, to examine supramaximal performance, and a 45-minute timed ride, to examine submaximal performance.

This study differed from the previous two in several significant ways.  For starters, peak power output declined in both groups, slightly more so in the high fat group (-10% vs. -8%).  Furthermore, RQ didn’t wasn’t significantly lower during this test in the high fat group, which possibly suggests they weren’t properly ketoadapted.  In Phinney’s study, the large energy deficit ensured ketoadaptation; this study lacked that aspect, somewhat more similar to Vogt’s, although unlike Vogt’s, these participants weren’t athletes which presumably makes ketoadaptation more difficult.

There are many factors at play… I wasn’t kidding when I said these studies are very nuanced!

Exhibit D. the infamous, Paoli 2012 

These were ‘elite artistic gymnasts,’ who could likely beat you in a race running backwards.  The ketogenic phase consisted of 55% fat and much more protein than the control phase (39% fat; protein: 41% vs. 15%). The significantly higher protein content was modestly offset by slightly more calories in the control phase, which reduces the amount of protein required to maintain nitrogen balance.

In this study, performance was, for the most part, ‘maintained,’ with relative increases in a few of the tests; eg, the “legs closed barrier.”  Changes in body composition were more robust: significantly reduced body fat and increased lean body mass after 30 days of ketogenic dieting (with their normal exercise routine).

Paoli data

The major confounder in this study was the use of an herbal cocktail only in the ketogenic diet group; despite this, the results are largely in line with the other studies.  For more on this study, see here.

Exhibit E. the most dramatic one to date: Sawyer 2013 

Please see here for the details, but in brief, strength-trained athletes showed improvements in high intensity exercise performance after only 7 days of carbohydrate restriction.  The nuances of this particular study are discussed more here.

barbell

Collectively, these studies show that physical performance in both endurance and high intensity realms does not always suffer, can be maintained, and in some cases is improved by ketogenic dieting.  Important factors are duration (to ensure adequate ketoadaptation), energy balance, and regular physical activity (athletes and regular exercisers can adapt to burning fat much quicker than sedentary folks).

 

calories proper

New study: high intensity exercise on a low carb diet.

Switch an athlete from their standard carbohydrate-rich diet to a low carb ketogenic one and suddenly performance tanks.  It is known.  Give them a few weeks to adapt, however, and it recovers.  This much was established for mainly endurance-related performance parameters by Steve Phinney and colleagues in the 1980’s (eg, Phinney et al., 1983).  Then, along came Antonio Paoli, Dominic D’Agostino, and others who showed a similar phenomenon in gymnasts, a population that routinely exercises at higher levels of intensity (Paoli et al., 2012).  Notably, in these studies the athletes were allowed adequate time to adapt to the new metabolic milieu – sometimes referred to as ketoadaptation.  Three weeks appears to be the minimum amount of time required for ketoadaptation; ie, studies of shorter duration generally show: low carb = poor physical performance.

…which is why I was surprised to see this one:

Effects of a short-term carbohydrate-restricted diet on strength and power performance (Sawyer et al., 2014)

These researchers subjected ~30 strength-trained individuals to a battery of performance assessments before and after 7 days of a low carb [ketogenic] diet.  Usually I would’ve stopped reading at this point because 7 days is too short.  But there were some nuances in the way this particular study was designed which piqued my interest.

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Advanced glycation end products (AGEs)

About a decade ago, Michael Brownlee posited that AGEs were one of The Four Horsemen responsible for the microvascular complications of diabetes.

Kill ‘em all

Thereafter, the image below (or a closely related one) appeared in at least one talk at every major diabetes conference for about 5 years.  Then it faded – maybe not because it is wrong, but rather just too simplistic to be useful (similar to CICO & ELMM).

Brownlee

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Skipping meals, intermittent fasting, grazing, etc.

or… Circadian Meal Timing!

They say if you’re going to [intentionally] skip a meal, it should be breakfast – and hey, that’s probably the easiest meal to skip.  However, a recent study showed skipping dinner FTW (well, not exactly).  I’ve never seen a proper study directly comparing the effects of skipping different meals, but here are a few that come close.  The findings may surprise you.

omelette

note: with the exception of Fernemark (Exhibit B), these studies are mostly macronutrient-controlled. That is, protein, fat, and carbs are similar between the groups; the only thing that differs is when they were ingested.  This can be tricky and/or very nuanced in some instances, like if dinner was smaller (fewer calories) but more protein-rich, for example… but in order to include 5 relevant studies and not bore you to death, you’ll have to check the full texts for those details.

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Nutrient Partitioning: …a *very* high protein diet.

Or: what happens when you eat a ton of protein?

RDA: 0.8 g/kg

Active individuals: 1.2-2.0 g/kg (via ISSN)
Comment (1): I think sedentary, physically inactive, and non-exercisers should be in this range to offset disuse atrophy.  And they should exercise.
Comment (2): Do athletes really need more protein than non-athletes?  They have exercise, a powerful anabolic stimulus.  More protein may improve performance or body composition, but they might not *need* it, in terms of nitrogen retention… there’s probably a study on this.

NEED =/= OPTIMIZATION

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Angiotensin: more than just blood pressure.

Pathologically low blood pressure can lead to shock & death.  Angiotensin II is there to prevent that, but it does much more.  A bit non-sequiter, perhaps.

This is what I call teamwork: low blood pressure detected by kidneys –> secretes renin.  Angiotensinogen (liver) is cleaved by renin to Angiotensin I.  Angiotensin Converting Enzyme (lungs [among other tissues]) cleaves angiotensin I into angiotensin II.

RAAS

Angiotensin II increases blood volume and restores blood pressure.  Good if you’ve lost a ton of blood fighting a wild beast; not good if you’re an overweight pen pusher on potato chips.  ACE inhibitors reduce angiotensin II, lowering blood pressure.  ACE is present in lungs probably because it deactivates bradykinin.  ACE inhibitors prevent this which might contribute to one of their side effects, a persistent dry cough which makes these drugs intolerable for many.  One alternative is angiotensin II receptor 1 blockers, or “ARBs.”


If anyone in pharma reads my blog (doubtful, unless they are monitoring for people to polonium-laced blow-dart), this will be their favorite post because I think ARBs are an interesting class of drugs.

If diet and weight loss are inadequate, telmisartan might be the next best thing to manage hypertension in diabetics:  Telmisartan for the reduction of cardiovascular morbidity and mortality (Verdecchia et al., 2011) –> effective at reducing mortality in patients with diabetes.

Efficacy of RAS blockers on cardiovascular and renal outcomes in NIDDM (Cae & Cooper 2012)  –> reduces morbidity and slows progression of renal disease (both hypertension and diabetes contribute to [irreversible] kidney damage, and frequently occur together, which makes this endpoint particularly relevant).  Hyperglycemia should be managed via diet, of course, and ARBs would need to be tested in people following something other than a Western diet (although said people may not even need treatment in the first place) (just thinking out loud here.  Or typing/whatever.)

But enough about blood pressure (<– boring); on to the more interesting stuff:

It started here: Chronic perfusion of angiotensin II causes cognitive dysfunctions and anxiety in mice (Duchemin et al., 2013)

Then: Candesartan prevents impairment of recall caused by repeated stress in rats (Braszko et al., 2012)

And: Anti-stress and anxiolytic effects of [candesartan] (Saavedra et al., 2005)

[Candesartan] prevents the isolation stress-induced decrease in cortical CRF1 receptor and benzodiazepine binding (Saavedra et al., 2006)

[Candesartan] ameliorates brain inflammation (Benicky et al., 2011)   brain inflammation induced by chronic exposure to artificial lights causes depression-like symptoms (in mice) (probably humans, too)

Finally, a human study: Candesartan and cognitive decline in older patients with hypertension (Saxby et al., 2008)

And then there’s this: Angiotensin receptor blockers for bipolar disorder (de Gois et al., 2013)


No mechanistic stuff because, well, I have no idea how it works.  On one hand, it might seem obvious that stress & anxiety can raise blood pressure, so something that lowers stress & anxiety could lower blood pressure.  Candesartan appears to do both (cause <–> effect?).  There are two unique properties of candesartan to note: 1) it gets into the brain; and 2) it leads to increased levels of angiotensin II (which presumably can’t do much because candesartan blocks the receptor for angiotensin II).  Perhaps angiotensin II targets a different receptor?  ARBs might blunt angiotensin II-induced CRH secretion, leading to anxiolysis, stress-tolerance, and pro-cognitive effects (that speculation was made possible by a thread on Avant Labs’ Forum and a few posts by Jane Plain on CRH [eg, here & here]).

Oh yeah, ARBs also prevent cafeteria diet-induced weight gain, insulin resistance, and ovulatory dysfunction [in rats] (Sagae et al., 2013).  And are sympatholytic like bromocriptine (Kishi & Hirooka 2013).

“The Angiotensin-melatonin axis” (Campos et al., 2013).

just sayin’

calories proper

Melatonin is the chemical expression of darkness.

Melatonin is secreted from the pineal gland, the seat of the soul, the third eye.   Pinealectomy induces circadian arrhythmia and has interesting effects on adipose tissue biology.

Exhibit A.  In 2004, Alonso-Vale and colleagues showed that 6 weeks after pinealectomy, [melatonin-deficient] rats subjected to fasting exhibited an impaired energy conservation response.  That is, they lost more weight and significantly depleted their adipocytes:

pinealectomy

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Pharmaceutical-grade circadian manipulation.

BMAL1 and CLOCK, ‘positive’ regulators of circadian gene expression, activate transcription of the negative regulators Per, Cry, and Rev-erb.  PER and CRY inhibit BMAL1 and CLOCK, whereas Rev-erb inhibits Bmal1.  It is said that Rev-erb is “an important link between the positive and negative loops of the circadian clock.”  You don’t really need to know any of that to follow this blog post.

circadian genes

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Diet study: American Diabetes Association vs. Low Carb Ketogenic

A randomized pilot trial of a moderate carbohydrate diet compared to a very low carbohydrate diet in overweight or obese individuals with type 2 diabetes mellitus or prediabetes (Saslow et al., 2014)

Disclaimer: this study was not ground-breaking; it was confirmation of a phenomenon that is starting to become well-known, and soon to be the status quo. That is, advising an obese diabetic patient to reduce their carb intake consistently produces better results than advising them to follow a low fat, calorie restricted diet.

The two diets:

Moderate carbohydrate diet: 45-50% carbs; 45 grams per meal + three 15 gram snacks = 165 grams per day; low fat, calorie restricted (500 Calorie deficit).  Otherwise known as a “low fat diet (LFD).”

In their words: “Active Comparator: American Diabetes Association Diet.  Participants in the American Diabetes Association (ADA) diet group will receive standard ADA advice. The diet includes high-fiber foods (such as vegetables, fruits, whole grains, and legumes), low-fat dairy products, fresh fish, and foods low in saturated fat.

Very low carbohydrate diet: Ketogenic; <50 grams of carb per day, no calorie restriction, just a goal of blood ketones 0.5 – 3 mM.

In their words: “Experimental: Low Carbohydrate Diet.  Participants will be instructed to follow a low carbohydrate diet: carbohydrate intake 10-50 grams a day not including fiber. Foods permitted include: meats, poultry, fish, eggs, cheese, cream, some nuts and seeds, green leafy vegetables, and most other non-starchy vegetables. Because most individuals self-limit caloric intake, no calorie restriction will be recommended.

Both groups were advised to maintain their usual protein intake.

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