I’ll zap the entire thingean

9 The effects of ten weeks of lower-body unstable

surface training on markers of athletic
performance.
Cressey EM, et al. J Strength Cond Res. 2007
May;21(2):561-7. [Pubmed]

11 Sorry, fructose... Red meat just stole the spotlight.

By Alan Aragon
Copyright © February 1st, 2012 by Alan Aragon
Home: www.alanaragon.com/researchreview
Correspondence: aarrsupport@gmail.com 14 D-Aspartic acid – finally, a legal alternative to
‘roids?
By Alan Aragon

2 Active joints are healthy joints.

By Matt Perryman 15 Chi L. Chiu comments on my review of Bray et al.

5 An investigative study into the influence of a

commercially available carbohydrate-protein-
electrolyte beverage on short term repeated
exercise performance.
Roberts JD, et al. J Int Soc Sports Nutr. 2012 Mar 9;9(1):5.
[Epub ahead of print] [Pubmed]

6 Sucrose-sweetened beverages increase fat

storage in the liver, muscle, and visceral fat depot:
a 6-mo randomized intervention study.
Maersk M, et al. Am J Clin Nutr. 2012 Feb;95(2):283-9.
Epub 2011 Dec 28. [Pubmed]

7 Different doses of supplemental vitamin D

maintain interleukin-5 without altering skeletal
muscle strength: a randomized, double-blind,
placebo-controlled study in vitamin D sufficient
adults.
Barker T, et al. Nutr Metab (Lond). 2012 Mar 9;9(1):16.
[Epub ahead of print] [Pubmed]

8 L-alanyl-L-glutamine ingestion maintains

performance during a competitive basketball
game.
Hoffman JR, et al. J Int Soc Sports Nutr. 2012 Mar 7;9(1):4.
[Epub ahead of print] [Pubmed]

Alan Aragon’s Research Review – February 2012 [Back to Contents] Page 1


Active joints are healthy joints.
By Matt Perryman
Editor’s note: Special thanks is due to Matt for contributing his
insight. A bounty of excellent information can be found at his
website, myosynthesis.com.
_______________________________________________
Introduction
I'm no stranger to injuries. Just about every joint in my body has
popped, ripped, or torn. If you exercise regularly, chances are
good that you've picked up a nagging ache or three, even if
you've never suffered a catastrophic injury.
In writing this article, my motivations have been selfish. Being
well acquainted with soft-tissue injuries, I've come to realize
how important it is to keep your joints and tendons healthy. You
can't be strong -- or fast, or anything else -- if you're hurt.
Managing injuries is an invaluable skill for a coach or athlete, so
I want to look at a wonderful paper from Michael Kjaer of
Denmark's University of Copenhagen:
Kjaer  M.  Role  of  extracellular  matrix  in  adaptation  of  tendon  whether that's lifting a limit barbell or landing from a jump.
and  skeletal  muscle  to  mechanical  loading.  Physiol  Rev.  2004  That's no easy feat, and tendons display an impressive range of
Apr;84(2):649‐98. [Pubmed]  both strength and spring-like stretching during the course of their
Dr. Kjaer is the head of the Muscle and Matrix group at the
University's Center for Healthy Aging, which means his team is
doing a lot of work on how muscles, and the support tissues
around muscles, adapt with age. While the focus is obviously on
aging, you have to understand how these tissues work under
normal conditions to see how they go wrong, so there's been a
lot of good work coming out of this group.
I found this paper so interesting because, firstly, it challenges
some ideas on recovery that we kick around in our gym-talk, and
also because these findings dovetailed nicely with my own
recent experiences in working around -- and through -- injuries.
This particular paper contains a startling amount of information
on soft tissues and how they adapt to exercise, clocking in at a
monstrous 50 pages, so I'm going to be giving you the barest-of-
bones overview with that in mind -- namely, why injuries
happen and what we can do about them.
Muscles and other stuff
When a muscle contracts, it acts somewhat like a rubber band.
Muscle differs from rubber bands in several key (and obvious)
ways, but the image of a stretchy, squishy elastic material makes
for a good analogy: when you stretch out a rubber band, it
tightens and will quickly snap back to its original length when
you let go of it. Rubber bands and muscles share this property of
viscoelasticity -- that is, their physical properties can change
depending on their length.
Muscle tissue can also contract under its own power, and these
contractions create active tension -- shortening rather than
Alan Aragon’s Research Review – February 2012
lengthening -- which pulls the bone along with it and causes
movement around a joint. That's Biomechanics 101.
Muscles and bones are exposed to extremely high forces, and
from avenues you might not expect. Forget weight training.
While 800 lb squats and 180 kilo clean & jerks look impressive,
you're exposed to far greater peak forces during your morning
jog. Even the simplest of playtime activities like running and
jumping involve surprisingly high, albeit brief, forces. With all
that stretching and rebounding going on, it takes a heavy-duty
adhesive to keep our muscles anchored to our bones.
Muscle tissue is wrapped up in a network of collagen and
support cells called the extracellular matrix (ECM), which works
as a scaffolding for cells to latch on to. The ECM also produces
growth factors and assorted cellular flavorings which keep these
tissues knit together. Where the muscles connect to bones, the
ECM thickens into a tough band of fibers called a tendon.
The collagen in the ECM makes up about 25% of the proteins in
your body, but you probably don't give your poor connective
tissues much thought (until the tendon in your shoulder shoots
searing pains through your arm). Tendons themselves have some
amazing properties, at least as interesting as muscle tissue.
Tendons work like a combination of a rope and a shock
absorber, absorbing and transmitting the forces of daily life,
job.
Active tissues
It wasn't all that long ago that connective tissues, and tendons in
particular, were thought to be little more than dead lumps of
meat. The last few decades have shown us that most biological
systems we thought to be passive lumps are really active, vocal
participants, signaling away with hormones and growth factors
and other assorted chemical messengers, and this includes our
connective tissues.
We all know that muscles respond to tension, which is why we
lift weights in the first place. Living tissues exist in a constant
state of protein turnover, with old and damaged proteins broken
down and replaced with freshly-synthesized siblings. Normally
these processes balance out, but during physical activity, when
stretching or contracting a muscle, we set off a cascade of
chemical signals within the muscle fiber. This barrage of signals,
in part due to the way proteins are broken down by contracting
and stretching, triggers an increase in protein synthesis that
eventually leads to a bigger, stronger muscle.
That's the rough picture of how our muscle tissues grow and
change in response to lifting weights. It turns out that tendons
respond to load-bearing exercise in the same way, by increasing
the rate of protein synthesis – in this case collagen – to offset the
increased rate of break down. The additional collagen proteins
thus reinforce the tendon, making it better able to handle high
loading.
Despite our tendency to zoom in on muscles and forget
everything else, this makes sense. Your athletic career wouldn't
[Back to Contents] Page 2
get very far if your muscles could suddenly lift 300 pounds but
your tendons, unchanged since you first picked up a weight,
could only handle 150. That would be like designing a car with
600 horsepower and leaving off the brakes. Connective tissues
grow and adapt along with your muscles (fascia-stretching myth
notwithstanding).
Since previous findings suggested tendons were basically dead,
it was assumed they were an avascular tissue, lacking any blood
supply. We now understand that tendons do receive a blood
supply, but the amount is paltry compared to the local muscle.
Kjaer notes that during exercise, a tendon receives only a
fraction of the blood flow of the nearby muscle:
"[I]t has been possible to demonstrate in human models that
blood flow within and around tendon connective tissue
increases up to sevenfold during exercise, both in young,
middle-aged, and elderly individuals...This increase is by far
smaller than the 20-fold increase in adjacent skeletal muscle
blood flow under similar exercise conditions."
Some blood does reach the tendon, but it's miniscule and seems
to be regulated by an entirely separate system. Kjaer adds that
tendons have a relatively low metabolic activity, so this tiny
amount is probably good-enough under normal circumstances,
but that might change during physical activity.
As we age, collagen accumulates advanced glycation end
products (AGEs -- yes, that's what they call them). These stiffer
"glycated" tendons can withstand higher stresses, but this has a
price. As Kjaer says, "the accumulation of AGEs with aging thus
indicates a stiffer and more load-resistant tendon and
intramuscular ECM structure, but on the other hand reduces the
ability to adapt to altered loading, as the turnover rate of
collagen is markedly reduced."
Aging, and injuries to the tendons which likewise accumulate
AGEs, makes your tendons stiffer and more resistant to high
loads, but reduces their ability to adapt to any damage. This can
be troublesome, as you'll realize if you've ever injured a tendon.
Overuse injuries
One of the reasons we don't push ourselves in training is overuse
injuries. The harder you go, and more often you go hard, the
higher your risk of injury. Tendons, like muscles, have a
threshold for activity and once you go beyond that, you're
risking problems like tendinitis or worse.
Kjaer notes that although we use the word "overuse" to describe
injuries that happen with repetitive loading, we don't actually
know that this is the case. Some injuries happen due to changes
in the collagen structure, while others involve the tendon
rubbing over bone and fraying like a rope. Overuse may
certainly be an issue, but it's not clear that loading per se is the
cause.
Countering the common belief, he lists several alternatives
which contribute to the development of overuse injuries. Most
important of these are tissue inflammation and the regulation of
blood flow and metabolism within the tissues.
Inflammation, the local immune response, has been a pet interest
of mine in recent months. Chemical messengers of
Alan Aragon’s Research Review – February 2012
inflammation, called cytokines, play a part in all manner of
nastiness, from depression to the feel-like-crap feelings you
experience the day after a nose-bleed workout. It seems that
tendon pain and overuse injuries are another thing we can add to
the list. Two cytokines, TNF-alpha and IL-1beta, directly act
against collagen synthesis, which we definitely don't want. Since
heavy training, especially unfamiliar or soreness-inducing
exercise, dramatically elevates these nasties, we can expect they
might contribute to the accumulation of tendon injuries by
hampering the process of tissue regeneration.
The inflammatory pathway COX-2 seems to have an effect on
tendon healing, as rats given anti-inflammatory drugs heal faster
from tendon injuries. Whether taking a good dose of NSAIDs
helps with overuse injuries is unknown, but prostaglandins --
inflammatory mediators produced by COX-2 -- seem to do bad
things for tendons. Kjaer suggests that anti-inflammatories,
which block the synthesis of prostaglandins, might have a
protective effect. This is an interesting result given that COX-2
and prostaglandins seem to have negative effects on protein
synthesis in muscles, at least at doses of 1000mg (though it
remains to be seen if this effect extends to actual hypertrophy, as
some studies show no differences in actual muscle-growth
outcomes on similar doses).
Lack of blood flow is a feature common in tendon injuries.
Hypoxia (lack of oxygen) triggers degenerative changes in
tendons, which jives with the reduced capillary density observed
in shoulder injuries. Less blood flow, and an overall decrease in
metabolic rate in the muscle, leads to broken-down tendons. The
way to solve this is more obvious: keep the thing moving.
Activity means blood flow and increased metabolic activity,
which means more protein turnover.
Pain is caused by the release of several nociceptive (pain-
causing) substances during both overuse and in plain old
mechanical loading, including the neurotransmitter glutamate.
Pain may or may not have anything to do with injury itself, so
your tendinitis may not be a cause for concern even if it is
aggravating. This, too, seems to trace back to inflammatory
signals, particularly the COX-2 and prostaglandin pathway.
Solutions
These findings get some ideas crackling. Kjaer writes that,
"several conservative treatments such as immobilization,
physical therapy, stretching, and pharmacological treatment
with NSAIDs as well as surgical procedures have by no
means produced impressive results."
Basically, the accepted treatments are barely better than nothing
at all, an observation which squares with my experience. I've
tried all these methods over the years, and while you eventually
get the pain under control, the problem is never fixed.
The pain can start all over again with the slightest aggravation,
and while the "don't do that" strategy might be sufficient for
some, it's not a solution compatible with any future athletic
goals.
Instead, Kjaer suggests we try some alternatives. He points out
that resistance exercise has been shown to improve overuse
injuries (though to be fair, that's talking "average folks" rather
[Back to Contents] Page 3
than lifters who may have caused an injury with weight
training). Loading "of a certain magnitude" combined with
stretching the area "induces increased reorganization of the
collagen structures" which may strengthen the tendon, and
eccentric loading seems to do this better than concentric.
That ties back to an old therapy I heard about years ago, and
which is my fall-back in case of tendinitis: you do high-rep
eccentric-only exercise for the afflicted area. Keep it up 2-3
times a week for a few weeks and your tennis elbow will feel
surprisingly better.
The big thing is to keep the hurt parts mobile, irrespective of
loading or anything else. Tendons receive a woeful amount of
blood while at rest, which means a reduced metabolic rate and
degenerative changes. Rest, while probably feeling good, only
lets the poor tendon rot away while starved of much-needed
blood. Keeping the area mobile and at least somewhat active
encourages blood flow and may encourage healing by way of
increased protein synthesis.
I'm going to ride the fence on the anti-inflammatory suggestion.
In my experience a nice dose of ibuprofen, say 800-1000mg,
does wonders for the soft-tissue pain especially while trying to
lift in agony, and this paper suggests that it might actually
accelerate the healing process. But large doses of ibuprofen,
especially taken often, come with their own risks. Other anti-
inflammatories may not be so brutal, but I have no experience
with them, so I have nothing to add. Your use of even OTC
drugs is a judgment call for you to make, so if this turns out to
be a bad idea, don't say I didn't warn you.
Kjaer's findings fit with my own on-going dealings with injuries.
Not only do the trouble spots seem to heal faster, and with less
chance of re-injury, when I keep them as active as I can, but I've
discovered that regular loading keeps everything feeling better.
The difference between squatting once or twice a week, versus
three or even five to six days a week, is unbelievable. You'd
expect that to increase the injury rate, and yet I've found exactly
the opposite. Mobility really is health. My shoulders, of all
things, seem to respond to benching 5-6 days a week far better
than they ever did with just one or two sessions.
This isn't a call for you to start training your injuries and nagging
tendon-aches under heavy weights every day of the week, but
you might want to consider putting them under some kind of
loading as often as you can. Get some blood in there, stretch it
out, and let it feel a little tension. Do some pushups, drag a sled
around, ride the exercycle, things like that would all qualify. In
Kjaer's words, healing from tendon overuse injuries "requires
adjusted loading rather than absence of loading in the form of
immobilization."
Staying active encourages growth and well-being, and if you've
got a pain that won't leave you alone, you might be better trying
to work through it rather than resting it and making the problem
worse.

Alan Aragon’s Research Review – February 2012 [Back to Contents] Page 4

 strength/power or hypertrophy-focused training. Furthermore,
the testing protocol was a single-day/repeated-bout endurance
An investigative study into the influence of a event, so its relevance could be limited to this type of scenario.
commercially available carbohydrate-protein-
electrolyte beverage on short term repeated exercise Comment/application
performance.
Roberts JD, et al. J Int Soc Sports Nutr. 2012 Mar 9;9(1):5.
[Epub ahead of print] [Pubmed]
BACKGROUND: The purpose of this study was to undertake an
independent investigation into the effects of ingesting a
carbohydrate-protein-electrolyte (CPE) beverage on repeated
submaximal and time-trial cycling performance. METHODS:
Sixteen recreationally trained males (height: 1.76 +/- 0.07 m;
weight: 70.05 +/- 7.90 kg; VO2max: 49.69 +/- 4.19 ml.kg1.min1)
performed two exercise trials separated by 7 days. Each trial
comprised two bouts of 90 minutes exercise separated by a 2 hour
recovery period. Each bout comprised 45 minutes exercise on a
cycle-ergometer at 60%VO2max (ST), followed immediately by a
45 minute performance test (PT). Participants were randomly
assigned an 8% CPE beverage or colour/taste matched placebo (PL)
prior to each trial. Participants consumed 100 ml of the assigned
beverage every 10 minutes during each ST, and 500 ml at 0 and 60
minutes into recovery (total caloric delivery per trial: 617.6 kcal for
CPE and12.8 kcal for PL). Mean power output (W), speed (km.hr1)
and distance covered (km) were assessed throughout both trials.
Expired air was sampled at 10 minute intervals throughout ST.
Blood glucose and lactate were assessed during ST and recovery.
RESULTS: Distance covered during ST was significantly reduced
with PL by 9.12% (20.18 +/- 0.28 km in ST1 v 18.34 +/- 0.36 km in
ST2; P = 0.0001). With CPE, distance covered, power output and
average speed were maintained between ST1 and ST2. Oxygen
uptake was not significantly different between ST1 and ST2, or As depicted above, the CPE beverage better preserved
conditions. Respiratory exchange ratio (RER) values decreased performance on the second 90-minute bout. It’s notable that both
from 0.98 +/- 0.02 in ST1 to 0.91 +/- 0.02 in ST2 for PL (P = groups consumed a protein bar (206 kcal, 21.6 g P, 17.0 g C, 5.7
0.003), supporting reduced total carbohydrate oxidation rates (P = g F), at the midpoint of the 2-hour recovery period between
0.007). Mean blood glucose was maintained in CPE across ST trials, bouts. The bar was given to “avoid any unnecessary risks of
and was significantly greater than PL in ST2 (4.77 +/- 0.09 severe hypoglycaemia occurring during the placebo trial.” It’s
mmol.L1 for CPE compared with 4.18 +/- 0.06 mmol.L1 for PL, P likely that without this bar, the performance decrements in the
< 0.001). Mean distance during PT2 was 2.96 km (or 17.1%) further
placebo condition would have been even greater.
with CPE than PL (P = 0.003). Mean power significantly decreased
across PT with PL (134.21 +/- 4.79 W and 106.90 +/- 3.25 W, Notice how in the first 90-minute bout, no ergogenic advantage
respectively; P < 0.04). CONCLUSIONS: The use of a CPE was seen in the CPE condition compared to non-caloric placebo
beverage improves short-term repeated exercise and subsequent in either the submaximal phase or the time trial phase. The
performance compared to PL. Higher rates of carbohydrate authors cited research with contrary results, where carbohydrate
oxidation, maintenance of plasma glucose, and decreased levels of consumed before & during 1-hour bouts enhanced performance
fatigue may be beneficial for secondary bouts of performance and beyond placebo.1,2 They speculated that the disparate results
faster recovery turnover. SPONSORSHIP: Maxinutrition Ltd. were due to a higher pre-exercise carbohydrate dose (67.1 g
versus 35.4 g in the present study).2
Study strengths
Keep in mind that the protocol in the present study was
From a conceptual standpoint, there’s still a relative scarcity of undergone by casual trainees. Desbrow et al saw no effect of
research examining the effect of carbohydrate-protein-electrolyte carbohydrate-electrolyte ingestion during a 1-hour time trial
(CPE) beverages on endurance performance, so any study in this done after a carefully controlled 24-hour period where well-
vein is a welcome addition. 72-hour food records were taken trained subjects were put on a diet designed to maximize
prior to each trial in order to control intake variability. endurance capacity.3 Another thing to keep in mind is that the
lack of performance difference in the first 90-minute bout was
Study limitations
seen despite the consumption of 617.6 kcal in the CPE beverage,
Subjects were “recreationally active,” so the results may or may versus 12.8 kcal in the placebo, so it’s not as if an insignificant
not apply to populations on the far ends of the continuum. Also, amount of fuel was provided. Therefore, while pre- &/or during-
the results could be limited to the endurance-focused testing exercise fueling can benefit exhaustive repeated-bouts, single
protocol, and might not necessarily be applicable to bouts lasting less than 2 hours might not reliably be enhanced.
Alan Aragon’s Research Review – February 2012 [Back to Contents] Page 5
Sucrose-sweetened beverages increase fat storage in
the liver, muscle, and visceral fat depot: a 6-mo
randomized intervention study.
Maersk M, et al. Am J Clin Nutr. 2012 Feb;95(2):283-9. Epub
2011 Dec 28. [Pubmed]
BACKGROUND: The consumption of sucrose-sweetened soft
drinks (SSSDs) has been associated with obesity, the metabolic
syndrome, and cardiovascular disorders in observational and
short-term intervention studies. Too few long-term intervention
studies in humans have examined the effects of soft drinks.
OBJECTIVE: We compared the effects of SSSDs with those of
isocaloric milk and a noncaloric soft drink on changes in total fat
mass and ectopic fat deposition (in liver and muscle tissue).
DESIGN: Overweight subjects (n = 47) were randomly assigned
to 4 different test drinks (1 L/d for 6 mo): SSSD (regular cola),
isocaloric semiskim milk, aspartame-sweetened diet cola, and
water. The amount of intrahepatic fat and intramyocellular fat
was measured with (1)H-magnetic resonance spectroscopy.
Other endpoints were fat mass, fat distribution (dual-energy X-
ray absorptiometry and magnetic resonance imaging), and
metabolic risk factors. RESULTS: The relative changes
between baseline and the end of 6-mo intervention were
significantly higher in the regular cola group than in the 3 other
groups for liver fat (132-143%, sex-adjusted
mean; P < 0.01), skeletal muscle fat (117-
221%; P < 0.05), visceral fat (24-31%; P <
0.05), blood triglycerides (32%; P < 0.01), and
total cholesterol (11%; P < 0.01). Total fat
mass was not significantly different between
the 4 beverage groups. Milk and diet cola
reduced systolic blood pressure by 10-15%
compared with regular cola (P < 0.05).
Otherwise, diet cola had effects similar to
those of water. CONCLUSIONS: Daily
intake of SSSDs for 6 mo increases ectopic fat
accumulation and lipids compared with milk,
diet cola, and water. Thus, daily intake of
SSSDs is likely to enhance the risk of
cardiovascular and metabolic diseases. This
trial is registered at clinicaltrials.gov as NCT00777647.
SPONSORSHIP: Supported by grants from The Danish
Council for Strategic Research, The Food Study Group/Danish
Ministry of Food, Agriculture and Fisheries, Novo Nordic
Foundation, and Clinical Institute at Aarhus University,
Denmark.
Study strengths
This the first controlled study to compare these 4 sociologically
relevant conditions (milk, regular soda, diet soda, water) over a
long-term period of 6 months. In addition to dual X-ray
absorptiometry (DXA) was used to measure gross body
composition, and H-magnetic spectroscopy was an extra step to
measure intrahepatic (liver) fat and intramuscular fat.
Accumulation of fat in muscle & liver is called ectopic fat
deposition. This was the first study to examine the effect of
beverages on ectopic fat deposition beyond 10 weeks.
Compliance checks and anthropometric measures were done
every 1.5 months. A standardized evening meal was consumed
the night before each test.
Alan Aragon’s Research Review – February 2012
Study limitations
The authors acknowledge that the main limitation of the study
was the small sample size, which compromised statistical power.
Additionally, the conditions were not blinded, so this could have
influenced the level of expectation bias (& thus behavior) of the
participants and the investigators. On average, the subjects were
36.7% body fat at 96.6 kg of total bodyweight. This clearly
indicates that this sample does not reflect the trained or
physically active population, who are far less susceptible to the
adverse effects of sugar-sweetened beverages. Along these lines,
the dose of the treatments was large enough to markedly limit its
applicability. Consuming 1 liter of regular soda per day (430
kcal, 107.5 g sugar) might be normal for certain populations (ie,
writers, software engineers, Walmart center-aisle shoppers), but
has little relevance to populations remotely oriented toward
fitness. Tracking of both dietary intake and physical activity
were minimal. A 7-day record of both of those variables was
submitted at the 3-month point, and again at the end.
Disappointingly, the authors did not specify the macronutrient
intake of any of the conditions. This is a major omission of
critical data for properly interpreting the results.
Comment/application
The main finding of this study – regular soda’s ability in large
doses to increase ectopic fat & visceral fat – was not too
surprising, based on the consistent correlations seen in previous
research.4-7 Other adverse effects of excess regular soda (its
fructose content in particular) were also predictable, such as
increased triglycerides and decreased insulin sensitivity.8,9 So,
for any sedentary person planning on drinking a liter of cola per
day, let this study be fair warning that it’s a bad idea.
Facetiousness aside, there definitely were interesting outcomes.
First off, changes in total fat mass & total bodyweight were not
significantly different between groups. This indicates a
compensatory decrease in energy intake in the milk & regular
soda groups. Secondly, contrary to recent concerns about the
weight gain potential of diet soda,10 its effects were virtually
identical to water. Perhaps the most interesting outcome was the
milk group’s marked decrease in subcutaneous & visceral fat,
despite bodyweight stability. No other groups lost fat from these
depots. This lends support to previous research suggesting that
dairy consumption preferentially reduces abdominal fat.11,12
[Back to Contents] Page 6

Different doses of supplemental vitamin D maintain
interleukin-5 without altering skeletal muscle strength:
a randomized, double-blind, placebo-controlled study
in vitamin D sufficient adults.
Barker T, et al. Nutr Metab (Lond). 2012 Mar 9;9(1):16. [Epub
ahead of print] [Pubmed]
BACKGROUND: Supplemental vitamin D modulates
inflammatory cytokines and skeletal muscle function, but results are
inconsistent. It is unknown if these inconsistencies are dependent on
the supplemental dose of vitamin D. Therefore, the purpose of this
study was to identify the influence of different doses of
supplemental vitamin D on inflammatory cytokines and muscular
strength in young adults. METHODS: Men (n = 15) and women (n
= 15) received a daily placebo or vitamin D supplement (200 or
4000 IU) for 28-d during the winter. Serum 25-hydroxyvitamin D
(25(OH)D), cytokine concentrations and muscular (leg) strength
measurements were performed prior to and during supplementation.
Statistical significance of data were assessed with a two-way (time,
treatment) analysis of variance (ANOVA) with repeated measures,
followed by a Tukey's Honestly Significant Difference to test
multiple pairwise comparisons. RESULTS: Upon enrollment, 63%
of the subjects were vitamin D sufficient (serum 25(OH)D [greater
than or equal to] 30 ng/ml). Serum 25(OH)D and interleukin (IL)-5
decreased (P < 0.05) across time in the placebo group. Supplemental
vitamin D at 200 IU maintained serum 25(OH)D concentrations and
increased IL-5 (P < 0.05). Supplemental vitamin D at 4000 IU
increased (P < 0.05) serum 25(OH)D without altering IL-5
concentrations. Although serum 25(OH)D concentrations correlated
(P < 0.05) with muscle strength, muscle strength was not changed
by supplemental vitamin D. CONCLUSION: In young adults who
were vitamin D sufficient prior to supplementation, we conclude
that a low-daily dose of supplemental vitamin D prevents serum
25(OH)D and IL-5 concentration decreases, and that muscular
strength does not parallel the 25(OH)D increase induced by a high-
daily dose of supplemental vitamin D. Considering that IL-5
protects against viruses and bacterial infections, these findings could
have a broad physiological importance regarding the ability of
vitamin D sufficiency to mediate the immune systems protection
against infection. SPONSORSHIP: This study was funded in part
the Deseret Foundation(Intermountain Healthcare, Salt Lake City,
UT USA) and by the ARUP Institute for Clinical and Experimental
Pathology (Salt Lake City, UT USA).
Study strengths
This is the first study to ever examine the effect of a low (200
IU) and high dose (4000 IU) of vitamin D (cholecalciferol; also
called D3) on circulating inflammatory cytokines and muscular
strength. The trial was randomized, double-blinded, and
placebo-controlled. Subject characteristics were very similar
between groups (<6% difference in body mass, <1% difference
in BMI).
Study limitations
The authors diligently acknowledge a number of limitations,
including a small sample size, short trial length, lack of physical
activity tracking, limited dosing protocols, insufficient frequency
of blood draws, variability across time in the cytokine data, and
the inherent limitations of the subject characteristics’
extrapolability to other populations such as smokers, obese,
critically ill, and elderly.
Alan Aragon’s Research Review – February 2012
Comment/application
The main findings were twofold: First, muscle strength (single-
leg peak isometric force & peak power output) was unaffected
by vitamin D supplementation. The authors’ sensationalistic
delivery is worth quoting (bolded emphasis by me):
“Additionally, despite concentrations correlating with muscle
strength, our shocking data reveal that muscular strength does
not parallel the increase in serum 25(OH)D concentrations
induced by supplemental vitamin D at 4000 IU/d.”
This outcome puts a damper on the hope that vitamin D
supplementation could be used as an ergogenic aid for
strength/power athletes & bodybuilders. Part of this momentum
is attributable to Pilz et al, who found that subjects taking 3332
IU of vitamin D3 for 1 year had statistically significant increases
in testosterone.13 Total testosterone increased from 10.7 to 13.4
nmol/l, and free testosterone levels increased from 0.222 to
0.267 nmol/l. Statistical significance is one thing, but clinical
relevance is another thing altogether. As it stands, the present
study does not support vitamin D supplementation as a special
agent of strength/size gains, despite hormonal implications.
Secondly, both doses of supplemental vitamin D prevented
serum 25(OH)D and interleukin-5 (IL-5) decreases in vitamin D
sufficient adults during the winter, but the higher dose (4000 IU,
right bar in the chart above) was not more effective than the
lower dose (200 IU, middle bar in the chart above) at preserving
IL-5 status.
There are a couple of inferences that can be drawn from this.
First off, more isn’t always better, since a low dose of vitamin D
actually outperformed a 20x higher dose for raising IL-5 levels.
The authors were stumped about how this happened. Secondly,
the rise in IL-5 could have important clinical implications. IL-5
stimulates the growth of B cells which aid in adaptive immunity
by forming antibodies that protect against pathogenesis. Rodent
cells incubated with 1,25(OH)D increased the expression of
GATA-3, which is a transcription factor that promotes IL-5
expression. Given this, the authors speculate that maintaining
serum vitamin D levels is necessary for the expression of
GATA-3, which in turn would prevent drops in IL-5. In doing
so, IL-5 would bolster immunoglobin production and protect
against a wide range of viruses and infectious diseases.
Vitamin D is one of the few supplements (like fish oil) that has
emerged as an effective agent for alleviating a broad range of
physiological concerns, as well as optimizing disease-free
conditions. The current evidence continues to mount in its favor,
but thus far, more still is not necessarily better.
[Back to Contents] Page 7
L-alanyl-L-glutamine ingestion maintains performance after an overnight fast, or what. An ideal design would involve a
during a competitive basketball game. standardized final evening meal and/or a standardized breakfast
before testing. The latter would more closely mimic real-life
Hoffman JR, et al. J Int Soc Sports Nutr. 2012 Mar 7;9(1):4.
conditions since competitive basketball players rarely if ever hit
[Epub ahead of print] [Pubmed]
the court on an empty stomach upon waking. No tracking or
BACKGROUND: The purpose of this study was to examine the reporting of dietary intake was mentioned at all. This leaves the
efficacy of L-alanyl-L-glutamine (AG) ingestion on basketball performance tests open to confounding variability.
performance, including jump power, reaction time, shooting
accuracy and fatigue. METHODS: Ten women (21.2 +/- 1.6 Comment/application
years; height: 177.8 +/- 8.7 cm; body mass: 73.5 +/- 8.0 kg), all
scholarship NCAA Division I basketball players, volunteered for
this study. Subjects participated in four trials, each consisting of
a 40-min basketball game with controlled time-outs for
rehydration. During the first trial (DHY) subjects were not
allowed to rehydrate, and the total weight lost during the contest
was used to determine fluid replenishment during the subsequent
three trials. During one trial subjects consumed only water (W),
while during the other two trials subjects consumed the AG
supplement mixed in water using either a low dose (1 g per 500
ml) (AG1) or high dose (2 g per 500 ml) (AG2) concentration.
All data assessed prior to and following each game were
converted into a [increment] score (Post results - Pre results). All
performance data were then analyzed using a one-way repeated
measures analysis of variance. RESULTS: During DHY
subjects lost 1.72 +/- 0.42 kg (2.3%) of their body mass. No
differences in fluid intake (1.55 +/- 0.43 L) were seen between
rehydration trials. A 12.5% (p = 0.016) difference in basketball
shooting performance was noted between DHY and AG1 and an
11.1% (p = 0.029) difference was seen between AG1 and W.
Visual reaction time was significantly greater following AG1 (p
= 0.014) compared to DHY. Differences (p = 0.045) in fatigue,
as determined by player loads, were seen only between AG2 and
DHY. No differences were seen in peak or mean vertical jump
power during any trial. CONCLUSION: Rehydration with AG
appears to maintain basketball skill performance and visual
reaction time to a greater extent than water only.
SPONSORSHIP: This study was supported by a grant from
The main findings were that compared to water, L-alanyl-L-
Kyowa Hakko USA, New York, NY.
glutamine (AG) significantly improved shooting accuracy (top
Study strengths chart) and visual reaction time. Fatigue, measured by a satellite-
based movement tracking system, was less in the higher-dose
The majority of sports supplementation & sports nutrition AG (AG2). Notably, the lower-dose AG (AG1) outperformed
studies to-date are done on men, so it’s refreshing to see an all- AG2 for enhancing shooting accuracy, and also for visual
female sample. The subjects were trained competitors, so this reaction time. This is another example of more not being better.
minimizes the chance for exaggerated & variable effects seen in As seen in the bottom chart, there was no significant difference
newbies. Hydration state prior to testing was controlled by in lower-body reaction time between AG and water. Predictably,
measuring urine specific gravity. A broad battery of functionally subjects performed consistently worse in dehydrated conditions
relevant tests were done, including performance, power, and compared to either water or AG conditions, with the exception
reaction tests. All tests were supervised by certified strength and of the vertical jump, which was not affected by any of the
conditioning specialists. Two dosing levels were tested, treatments. Interestingly, this lack of effect happened despite a
providing useful data for establishing potential effectiveness loss of 2.3% of total bodyweight during the game.
thresholds & dose-response relationships. A crossover was done
The authors speculate that AG may have increased the efficiency
so that each subject underwent each treatment, minimizing the
of fluid uptake from the gut, thereby minimizing the ergolytic
confounding variability of inter-individual response.
effects of dehydration on nerve conduction and brain function.
Although AG’s results in this study were not all that impressive,
Study limitations
they nonetheless support previous research by Hoffman et al,
The sample was small (10 subjects), but as mentioned, this showing that AG increased time to exhaustion during mild
inherent compromise in statistical power was partially alleviated hypohydration.14 It’s worth mentioning that the AG research
by the crossover. Nowhere in the manuscript was the time of thus far has been sponsored by the maker of Sustamine, the
testing reported. There’s no indication of whether it was done brand name of the AG product tested in this study.

Alan Aragon’s Research Review – February 2012 [Back to Contents] Page 8

 Comment/application

The effects of ten weeks of lower-body unstable

surface training on markers of athletic performance.
Cressey EM, et al. J Strength Cond Res. 2007 May;21(2):561-7.
[Pubmed]
BACKGROUND: Initially reserved for rehabilitation programs,
unstable surface training (UST) has recently grown in popularity in
strength and conditioning and general exercise scenarios.
Nonetheless, no studies to date have examined the effects of UST
on performance in healthy, trained individuals. PURPOSE: The
purpose of this study was to determine the effects of 10 weeks of
lower-body UST on performance in elite athletes. DESIGN:
Nineteen healthy, trained members (ages 18-23 years) of a National
Collegiate Athletic Association Division I collegiate men's soccer
team participated. The experimental (US) group (n = 10)
supplemented their normal conditioning program with lower-body
exercises on inflatable rubber discs; the control (ST) group (n = 9)
performed the same exercises on stable surfaces. Bounce drop jump
(BDJ) and countermovement jump (CMJ) heights, 40- and 10-yard
sprint times, and T-test (agility) times were assessed before and
after the intervention. RESULTS: The ST group improved
significantly on predicted power output on both the BDJ (3.2%) and
CMJ (2.4%); no significant changes were noted in the US group.
Both groups improved significantly on the 40- (US = -1.8%, ST = -
3.9%) and 10-yard sprint times (US = -4.0%, ST = -7.6%). The ST
group improved significantly more than the US group in 40-yard
As seen above, stable training significantly improved the
sprint time; a trend toward greater improvement in the ST group
performance of jumping, while unstable training did not (Table
was apparent on the 10-yard sprint time. Both groups improved
2). Both improved for 40-yard sprinting, but stable training did
significantly (US = 2.9%, ST = -4.4%) on T-test performance; no
so to a significantly greater degree (Table 3). Stable training was
statistically significant changes were apparent between the groups.
also better at improving 10-yard sprinting, but not to a degree of
CONCLUSION: These results indicate that UST using inflatable
rubber discs attenuates performance improvements in healthy,
statistical significance. Agility improved significantly in both
trained athletes. Such implements have proved valuable in
groups, with neither showing a significant advantage. It’s
rehabilitation, but caution should be exercised when applying UST notable that for jumping & sprinting, stable training did not
to athletic performance and general exercise scenarios. merely outperform unstable training, but the latter actually
SPONSORSHIP: None listed. appeared to prevent improvements from occurring in these
particular tests.
Study strengths At least two studies comparing stable versus unstable training
have been published after the present one.15,16 Both showed
This was the first study to ever compare the effects of stable
equal effectiveness of each type for improving performance. But
versus unstable surface training in trained athletes with no recent
unlike the present study, the testing included direct challenges to
history of injury. All subjects were familiar & proficient with
balancing ability, and used untrained subjects, who are more
each of the tests, which reduced the confounding potential of
prone to benefit from a wider variety of stimuli. The more highly
learning effects. The training protocols were designed to be
trained someone is, the greater the need to match the training
identical in volume. The unstable training was deferred to the
stimuli with the intended adaptations.
end of the training bouts. This better reflects its use in real-world
protocols, where it’s typically implemented as assistance work The authors of the present study speculate that unstable training
rather than primary work. can undermine the specificity of training by hindering
adaptations of activity that occurs on stable surfaces (which
Study limitations includes most major sports). Although unstable training might
benefit individuals with injuries or compromised proprioception,
6 of the 10 weeks were offseason conditions while the final 4 it can also impair the stretch-shortening cycle through both
weeks coincided with the subjects’ competitive season. This mechanical & psychological means. This quote sums it up well:
could have introduced rogue variables. It would have been more
ideal from a control standpoint if a single/static set of conditions “Effectively, by training slowly and tentatively, the athlete may
was maintained through the full length of the trial. Sample size be conditioned to perform in the same slow manner when faced
was small (10 subjects in the unstable group, 9 in the stable with athletic challenges. Antagonist activity is heightened during
group). Dietary intake – both in general, and relative to the tests UST to maintain joint stability, so it is not unreasonable to
– was not mentioned at all in the manuscript. This opens the conjecture that such a training effect could be detrimental to
possibility for confounding variability, since nutrition is an optimal rate and magnitude of force production when applied
important factor in performance testing. for an extended training period.”
Alan Aragon’s Research Review – February 2012 [Back to Contents] Page 9
1. el-Sayed MS, et al. Carbohydrate ingestion improves
endurance performance during a 1 h simulated cycling time
trial. J Sports Sci. 1997 Apr;15(2):223-30. [Pubmed]
2. Rollo I, Williams C. Influence of ingesting a carbohydrate-
electrolyte solution before and during a 1-hr running
performance test. Int J Sport Nutr Exerc Metab. 2009
Dec;19(6):645-58. [Pubmed]
3. Desbrow B, et al. Carbohydrate-electrolyte feedings and 1 h
time trial cycling performance. Int J Sport Nutr Exerc
Metab. 2004 Oct;14(5):541-9. [Pubmed]
4. Nseir W, et al. Soft drinks consumption and nonalcoholic
fatty liver disease. World J Gastroenterol. 2010 Jun
7;16(21):2579-88. [Pubmed]
5. Lim JS, et al. The role of fructose in the pathogenesis of
NAFLD and the metabolic syndrome. Nat Rev
Gastroenterol Hepatol. 2010 May;7(5):251-64. Epub 2010
Apr 6. [Pubmed]
6. Abid A, et al. Soft drink consumption is associated with
fatty liver disease independent of metabolic syndrome. J
Hepatol. 2009 Nov;51(5):918-24. Epub 2009 Aug 21.
[Pubmed]
7. Assy N, et al. Soft drink consumption linked with fatty liver
in the absence of traditional risk factors. Can J
Gastroenterol. 2008 Oct;22(10):811-6. [Pubmed]
8. Stanhope KL. Role of fructose-containing sugars in the
epidemics of obesity and metabolic syndrome. Annu Rev
Med. 2012;63:329-43. Epub 2011 Oct 27. [Pubmed]
9. Stanhope KL, et al. Consumption of fructose and high
fructose corn syrup increase postprandial triglycerides,
LDL-cholesterol, and apolipoprotein-B in young men and
women. Clin Endocrinol Metab. 2011 Oct;96(10):E1596-
605. Epub 2011 Aug 17. [Pubmed]
10. Fowler SP, et al. Fueling the obesity epidemic? Artificially
sweetened beverage use and long-term weight gain. Obesity
(Silver Spring). 2008 Aug;16(8):1894-900. Epub 2008 Jun
5. [Pubmed]
11. Poddar KH, et al. -fat dairy intake and body weight and
composition changes in college students. J Am Diet Assoc.
2009 Aug;109(8):1433-8. [Pubmed]
12. Zemel MB. The role of dairy foods in weight management.
J Am Coll Nutr. 2005 Dec;24(6 Suppl):537S-46S. [Pubmed]
13. Pilz S, et al. Effect of vitamin D supplementation on
testosterone levels in men. Horm Metab Res. 2011
Mar;43(3):223-5. Epub 2010 Dec 10. [Pubmed]
14. Hoffman JR, et al. Examination of the efficacy of acute L-
alanyl-L-glutamine ingestion during hydration stress in
endurance exercise. J Int Soc Sports Nutr. 2010 Feb 3;7:8.
[Pubmed]
15. Sparkes R, Behm DG. Training adaptations associated with
an 8-week instability resistance training program with
recreationally active individuals. J Strength Cond Res. 2010
Jul;24(7):1931-41. [Pubmed]
16. Kibele A, Behm DG. Seven weeks of instability and
traditional resistance training effects on strength, balance
and functional performance. J Strength Cond Res. 2009
Dec;23(9):2443-50. [Pubmed]

Alan Aragon’s Research Review – February 2012 [Back to Contents] Page 10


Sorry, fructose... Red meat just stole the spotlight.
By Alan Aragon
_________________________________________________________________
Introduction
A recent study by Pan et al has caused more panic and chaos
than I’ve seen in a very long time.1 I received plenty of requests
for my take on it, and was tempted to blog about it. But let’s face
it – I can give a much nerdier analysis here, without having to
field comments from every walk (& crawl) of life. Several
health/fitness writers have already commented on the study, and
I was surprised to see that no one has yet discussed the data
collection limitations or the counter-evidence to any degree of
depth. In the following discussion, I’ll look at the study’s key
strengths, limitations, & related research. But first, here’s the
abstract to provide a snapshot of the details:
Strengths
It’s important to note that this study had several design strengths,
although the weight of each strength is considerably less than
that of its limitations, which I’ll get to. A common strength of
epidemiological studies is their ability to examine massive
groups of people (in this case a total of 121,342 subjects) over
long periods of time (in this case 28 years, collectively).
Interventional designs can rarely come close to epidemiological
research in terms of trial duration & sample size due to the
financial and logistical constraints involved with maintaining
control of all the necessary variables.
Extra measures were taken in attempt to suppress confounding
variables. The authors conducted sensitivity analyses in order to
account/adjust for what they felt were the major dietary & non-
dietary variables. Efforts were made to correct for measurement
error, using energy density of red meat intake instead of crude
intake (which ignores the food’s context within total calories
consumed).

Red  Meat  Consumption  and  Mortality:  Results  From  2  Prospective studies were examined, which means that they were
Prospective Cohort Studies.  designed prior to any data collection. Retrospective designs
Pan A, et al. Arch Intern Med. 2012 Mar 12. [Epub ahead of print]   would have been more highly subject to broken, inconsistent, or
[Pubmed]  irrelevant data sets. Along these lines, the authors mentioned that
  their study had “high rates of long-term follow-up, and detailed
BACKGROUND:  Red  meat  consumption  has  been  associated 
with  an  increased  risk  of  chronic  diseases.  However,  its  and repeated assessments of diet and lifestyle.” While this might
relationship  with  mortality  remains  uncertain.  PURPOSE:  We  be true compared to retrospective designs, these control
prospectively  observed  37  698  men  from  the  Health  measures have limited reach – as we’ll see in the next section.
Professionals Follow‐up Study (1986‐2008) and 83 644 women 
from  the  Nurses'  Health  Study  (1980‐2008)  who  were  free  of  Limitations
cardiovascular  disease  (CVD)  and  cancer  at  baseline.  Diet  was  The fundamental limitation of epidemiological research is that
assessed  by  validated  food  frequency  questionnaires  and  it’s observational rather than interventional. As such, there is no
updated every 4 years. DESIGN: We prospectively observed 37  true control of the variables. There’s no direct suppression or
698 men from the Health Professionals Follow‐up Study (1986‐
elimination of superfluous factors that can interfere with the
2008) and 83 644 women from the Nurses' Health Study (1980‐
isolation of the causal agents and their effects.
2008)  who  were  free  of  cardiovascular  disease  (CVD)  and 
cancer  at  baseline.  Diet  was  assessed  by  validated  food  Observational research (which includes cohort, cross-sectional,
frequency questionnaires and updated every 4 years. RESULTS:  and case-control studies) is limited to drawing correlational or
We documented 23 926 deaths (including 5910 CVD and 9464  associative relationships. They are good for providing hints,
cancer  deaths)  during  2.96  million  person‐years  of  follow‐up.  questions, and food for thought, but not firm conclusions.
After  multivariate  adjustment  for  major  lifestyle  and  dietary  Observational research is useful for generating hypotheses that
risk  factors,  the  pooled  hazard  ratio  (HR)  (95%  CI)  of  total  can then be subjected to the more rigorous investigation in
mortality for a 1‐serving‐per‐day increase was 1.13 (1.07‐1.20)  Randomized Controlled Trials (RCTs). RCTs are considered the
for  unprocessed  red  meat  and  1.20  (1.15‐1.24)  for  processed  gold standard of research design because it’s the only type of
red  meat.  The  corresponding  HRs  (95%  CIs)  were  1.18  (1.13‐ research that can establish cause-and-effect relationships.
1.23)  and  1.21  (1.13‐1.31)  for  CVD  mortality  and  1.10  (1.06‐
1.14)  and  1.16  (1.09‐1.23)  for  cancer  mortality.  We  estimated  Whenever a headline contains the phrase “associated with,”
that substitutions of 1 serving per day of other foods (including  “correlated with,” “linked to,” or “increases the risk of,” you
fish, poultry, nuts, legumes, low‐fat dairy, and whole grains) for  can bet it’s based on observation rather than controlled
1  serving  per  day  of  red  meat  were  associated  with  a  7%  to  intervention. Observational research makes headlines more often
19%  lower  mortality  risk.  We  also  estimated  that  9.3%  of  than interventional research because their long durations & large
deaths  in  men  and  7.6%  in  women  in  these  cohorts  could  be  subject numbers can track the incidence of emotionally charged
prevented  at  the  end  of  follow‐up  if  all  the  individuals  topics like death and disease. The problem is, observational
consumed  fewer  than  0.5  servings  per  day  (approximately  42  research can be plagued with multiple rogue variables. As such,
g/d)  of  red  meat.  CONCLUSION:  Red  meat  consumption  is  epidemiology is more prone than controlled experiments to
associated  with  an  increased  risk  of  total,  CVD,  and  cancer  suggesting relationships between variables that happened in spite
mortality. Substitution of other healthy protein sources for red  of each other – not because of each other.
meat is associated with a lower mortality risk. SPONSORSHIP: 
This  study  was  supported  by  grants  from  the  National  Aside from these fundamental limitations of observational
Institutes  of  Health  and  the  National  Heart,  Lung,  and  Blood  research, the data collection method can profoundly influence
Institute (Dr Sun).  the validity of the outcomes. The authors assert that the use of

Alan Aragon’s Research Review – February 2012 [Back to Contents] Page 11

food frequency questionnaires (FFQs) to record dietary intake sounds like a picture-perfect recipe for increasing the risk for
data were a design strength. In reality, FFQs are the most early death & disease. This helps illustrate my point that
commonly used means of intake tracking in epidemiology observational research often finds things guilty by mere
because it’s an inexpensive and simple way to collect this data. association rather than direct effects.
Rather than hiring professionals to communicate with subjects
An important methodological slip was the lack of proper
directly, subjects can fill out FFQs themselves. This is a quicker,
delineation between processed and unprocessed red meat.
easier process than maintaining & analyzing ongoing food
Questionnaire items with the unprocessed designation included
records. However, it lacks rigor and is highly subject to error.
hamburger. There’s a wide range of contexts (healthy & not)
The authors tout the frequency & regularity of FFQ follow-ups, that hamburger can be consumed in. However, there likely
but considering that this was done every four years, a huge would be a significant, unaccounted incidence of consumption
amount of unchecked variability is possible. To illustrate these of hamburger within fast food, which often is accompanied by a
limitations, Marks et al found that compared to weighed food host of highly refined, calorically dense foods.
records, FFQs overestimated the intake of most foods,
When proposing possible mechanisms for red meat’s adverse
particularly vegetables & fruits.2 In fact, they found that FFQ-
potential, the authors first point to its saturated fat and
reported vegetable intake was about twice the amount reported
cholesterol content as contributors to cardiovascular disease risk.
in weighed food records. In another example, Schaefer et al
But ironically, in both cohorts studied, there was an inverse
found that FFQs significantly underestimated intakes of intakes
relationship between blood cholesterol levels and red meat
of dietary fats, cholesterol, and protein. 3 In addition, FFQs failed
intake (see the table here). In other words, those who consumed
to reliably estimate carbohydrate intake within a high-fat diet,
the most red meat had the lowest cholesterol levels. This directly
and fat intake within a very low-fat diet. Diet records were found
contradicts the authors’ speculations that reflect the outdated
to be more accurate and reliable than FFQs.
outlook influenced by the lipid hypothesis.
The lack of FFQ reliability becomes particularly visible when
cross-checked with the doubly labeled water (DLW) technique, Recent counter-evidence
which is considered the gold standard of measuring energy
A systematic review & meta-analysis by Micha et al examined
expenditure in free-living conditions.4 DLW is biomarker-based,
the data from 20 studies (17 prospective cohort, 3 case-control)
which makes its accuracy independent of self-reported intake
which collectively contained 1,218,380 subjects.8 They found
error. Therefore, DLW can objectively enable the detection of
that consumption of processed meats, but not red meats, was
reporting bias.5 For example, Mahabir et al found that in
associated with a higher incidence of coronary heart disease and
postmenopausal women, FFQ data underestimated total energy
diabetes. Notably, most of the processed meats were red meats.
intake compared to total energy expenditure assessed via DLW
The authors acknowledge that all the studies in their analysis
by a whopping 42%.6 In another example, Schatzkin et al found
were observational, so the erroneous influence of unaccounted
that when cross-checked with DLW, FFQ was found to have
variables cannot be dismissed.
considerably low validity.7 To quote them,
“Our results indicate that the FFQ cannot be recommended So, what about the more rigorous data from RCTs? A very
as an instrument for evaluating the absolute intakes of energy recent study by Roussel et al examined the effect of varying
and protein in relation to disease.” amounts of lean beef on blood lipids, lipoproteins, and
apolipoproteins of hypercholesterolemic subjects.9 Unlike the
Since recall-based, self-reported data is notoriously inaccurate, subjects of Pan et al, who were likely to be on various unhealthy
the use of FFQs is best viewed as a necessary evil rather than a incarnations of the standard Western diet, Roussel et al’s
design strength. And since the reliability of the data hinges upon subjects were on regimes designed to improve blood lipid
the accuracy of the collection method, any compromise in this profiles. The diets were described as, “rich in fruit, vegetables,
methodology inevitably compromises the validity of the results. and lean meats consistent with food-based dietary
Aside from the inability of observational research to demonstrate recommendations.” Here are the vital stats of each diet:
causation, and the shaky reliability of FFQs, there were a few
other results that challenge the authors’ conclusions. Although
red meat (more so for processed than unprocessed red meat) was
found to be culpable for adverse outcomes even after adjusting
for multiple dietary & non-dietary factors, the following findings
stand out like a sore thumb:
“Men and women with higher intake of red meat were less
likely to be physically active and were more likely to be
current smokers, to drink alcohol, and to have a higher body
mass index. In addition, a higher red meat intake was
associated with a higher intake of total energy but lower
intakes of whole grains, fruits, and vegetables.”
Excessive smoking, drinking, and total caloric intake – coupled
with a lack of physical activity and lower intake of plant foods

Alan Aragon’s Research Review – February 2012 [Back to Contents] Page 12

Note that the HAD diet served as a control diet, containing more
total & saturated fat, and less fiber than the other diets. Here are
the blood lipid results of the 5-week intervention:
The DASH, BOLD, & BOLD+ contained 28 g, 113 g, & 153 g
(1, 4, & 5.4 oz) lean beef per day. Nevertheless, as seen above,
all of the lean beef-containing diets had a significantly greater
blood cholesterol-lowering effect than HAD, with no significant
differences between groups. Interestingly, a nonsignificant rise
in triacylglycerol (TG) in the DASH diet, which contained the
least beef. The strongest TG decrease was seen in the BOLD+
diet, with the highest beef dose. But this is unsurprising since it
had a higher protein & lower carbohydrate content than the rest
of the diets. In addition, BOLD+ was the only diet that
significantly lowered the atherogenic apolipoprotein B compared
to the control diet. The take-home message of this study is that
when lean beef is included within the context of a sensible diet
(unlike what largely was examined in Pan et al’s observational
study), health can actually improve. Amazing how context
actually matters. To quote the authors’ concluding statement:
“The results of the BOLD study provide convincing evidence
that lean beef can be included in a heart-healthy diet that
meets current dietary recommendations and reduces CVD
risk.”
There are a couple of things I’d like to add. First off, these
improvements were seen despite the inclusion of a formal
exercise regimen, which undoubtedly would have improved
health indexes even further. And these improvements would
likely be most amplified in the BOLD+ diet if resistance training
was included in the training regimen. Another noteworthy tidbit
is that carbohydrate was the dominant macronutrient in all the
diets, with fat in the middle, and protein in the smallest
proportion. It’s important to note that the current evidence is not
necessarily in favor of carbohydrate-dominant diets for lowering
cardiovascular disease risk, at least in sedentary & non-athletic
populations. A strong illustration is a systematic review of RCTs
by Hession et al, who concluded that low-carbohydrate/high-
protein diets are at least equally effective, if not more so than
low-fat/high-carbohydrate diets for reducing bodyweight and
cardiovascular disease risk.10
Conclusions
The observational data indicting red meat – especially
unprocessed meat – is weak compared to the evidence from
Alan Aragon’s Research Review – February 2012
RCTs showing that red meat has neutral-to-beneficial effects on
health. The main caution with consuming fattier cuts of red meat
rather than leaner cuts is the potential impingement upon total
caloric allotment. But as long as this target isn’t breached, there
simply isn’t enough compelling evidence to avoid or minimize
unprocessed red meat intake.
Even in the case of processed red meats, consumption in
moderation (let’s say, 2-3 servings a week), along with regular
intake of vegetables, fruits, whole grains, legumes, and a variety
of fat sources (including omega-3-rich sources) is not likely to
raise any health risks, especially in normal-weight, physically
active populations. Ultimately, Pan et al’s red meat study is not
completely worthless. It fact, it contains the hidden message that
excessive eating, smoking, drinking, and sitting can hinder the
goal to be healthy.
References
1. Pan A, et al. Red Meat Consumption and Mortality: Results
From 2 Prospective Cohort Studies. Arch Intern Med. 2012
Mar 12. [Epub ahead of print] [Pubmed]
2. Marks GC, et al. Relative validity of food intake estimates
using a food frequency questionnaire is associated with sex,
age, and other personal characteristics. J Nutr. 2006
Feb;136(2):459-65. [Pubmed]
3. Schaefer EJ, et al. Lack of efficacy of a food-frequency
questionnaire in assessing dietary macronutrient intakes in
subjects consuming diets of known composition. Am J Clin
Nutr. 2000 Mar;71(3):746-51. [Pubmed]
4. Plasqui G, Westerterp KR. Physical activity assessment
with accelerometers: an evaluation against doubly labeled
water. Obesity (Silver Spring). 2007 Oct;15(10):2371-9.
[Pubmed]
5. Trabulsi J, Schoeller DA. Evaluation of dietary assessment
instruments against doubly labeled water, a biomarker of
habitual energy intake. Am J Physiol Endocrinol Metab.
2001 Nov;281(5):E891-9. [Pubmed]
6. Mahabir S, et al. Calorie intake misreporting by diet record
and food frequency questionnaire compared to doubly
labeled water among postmenopausal women. Eur J Clin
Nutr. 2006 Apr;60(4):561-5. [Pubmed]
7. Schatzkin A, et al. A comparison of a food frequency
questionnaire with a 24-hour recall for use in an
epidemiological cohort study: results from the biomarker-
based Observing Protein and Energy Nutrition (OPEN)
study. Int J Epidemiol. 2003 Dec;32(6):1054-62. [Pubmed]
8. Micha R, et al. Red and processed meat consumption and
risk of incident coronary heart disease, stroke, and diabetes
mellitus: a systematic review and meta-analysis.
Circulation. 2010 Jun 1;121(21):2271-83. Epub 2010 May
17. [Pubmed]
9. Roussell MA, et al. Beef in an Optimal Lean Diet study:
effects on lipids, lipoproteins, and apolipoproteins. Am J
Clin Nutr. 2012 Jan;95(1):9-16. Epub 2011 Dec 14.
[Pubmed]
10. Hession M, et al. Systematic review of randomized
controlled trials of low-carbohydrate vs. low-fat/low-calorie
diets in the management of obesity and its comorbidities.
Obes Rev. 2009 Jan;10(1):36-50. Epub 2008 Aug 11.
[Pubmed]
[Back to Contents] Page 13

D-Aspartic acid – finally, a legal alternative to ‘roids?
By Alan Aragon
 
             It  seems   like   the  entire  supplement  forum  is  on  D‐  
             Aspartic  acid.  Does  it  really  work  for  boosting  test  In the non-training group, total & free testosterone levels at
levels? I’d love to hear your take on it, thanks.   baseline were 502 ng/dl & 79 pg/ml, respectively. At 10 weeks
Let me get a little background out of the way first. D-
Aspartic acid (D-Asp) is a naturally occurring amino acid
in neuroendocrine tissues.1 Rodent studies have shown that D-
Asp is involved in the production of luteinizing hormone (LH)
and testosterone, and has recently been shown to function as a
neurotransmitter.2 D-Asp is likely to play an important role in
reproduction, based on its involvement in the synthesis of sex
hormones.
A single human study with borderline relevance
Despite an abundance of animal research, D-asp has one in vivo
human study examining its effect on serum androgen levels. In a
12-day trial on healthy men aged 27-37, Topo et al examined the
effect of a 3.12 g daily oral dose of D-Asp.3 LH levels went from
4.2 to 5.6 mIU/m, which is a 33.3% increase. Three days after
the cessation of D-Asp dosing, LH was still elevated 14% above
baseline levels. Testosterone levels rose from a mean value of
4.5 to 6.4 ng/ml, which is a 42% increase. Three days after
cessation, testosterone was still elevated 22% above baseline
levels. This residual effect suggests that D-Asp accumulates in
the testes during regular dosing.
Topo et al’s study is the most significant piece of evidence that
consumers and sports supplement sellers stake their hopes and
claims upon. However, the study has some important limitations.
First of all, the 12-day duration was very short, telling us next to
nothing about whether androgen levels would continue to rise or
not. The short duration also prevents any assessment of D-Asp’s
effect on practical endpoints like exercise performance or body
composition, which are the main interests of the fitness &
bodybuilding audience. In addition, 12 days is far too little time
to properly test for safety and side effects. A final limitation is
the neglect to measure free testosterone. They only reported total
testosterone, which tells us little about D-Asp’s effect on the
more bioavailable form of testosterone.
Comparison with supraphysiological dosing
The hope of many consumers is that D-Asp supplementation will
have effects similar to anabolic-androgenic steroids (AAS).
However, there’s a widespread misunderstanding of the
difference between the effect of pharmacological doses of AAS
compared to the D-Asp effects seen in Topo et al. Despite the
42% increase in total testosterone, 6.4 ng/ml (640 ng/dl) is still
within normal physiological limits. Normal values in males can
range roughly 300-1200 ng/dl, depending on age and other
factors.4,5 Therefore, while the increase seen by Topo et al
reached statistical significance, it lacks clinical relevance.
A classic example of the powerful effects of supraphysiological
AAS dosing is a well-designed study by Bhasin et al, where men
Alan Aragon’s Research Review – February 2012
aged 19-40 were given intramuscular injections of testosterone
enanthate dosed at 600 mg/week for 10 weeks.6 Substantial
muscular size and strength gains were seen in both the resistance
training and non-training groups on testosterone. The rate of lean
mass gain in the testosterone + training group was roughly
double that typically seen in creatine supplementation research.
of testosterone treatment, those levels hit 2828 ng/dl & 497
pg/ml, respectively. In the resistance training group, total & free
testosterone levels were 431 ng/dl & 90 pg/ml, respectively. 10
weeks of testosterone treatment raised those values to 3244 ng/dl
& 572 pg/ml, respectively. To put this into perspective, D-Asp
caused a 42% increase in testosterone, whereas a
supraphysiological AAS dose increased testosterone by 563% in
non-training subjects, and 752% in subjects on a strength
training program.
600 mg/week is a relatively high testosterone dose, especially in
circles outside of competitive sports. However, much lower
doses have been beneficial for individuals with abnormally low
testosterone. For example, Bhasin et al found that in
hypogonadal males aged 19-47, a replacement dose of 100
mg/week for 10 weeks significantly increased muscle mass and
strength.7 Baseline total & free testosterone levels were 2.5
nmol/l (72 ng/dl) & 66 pmol/l, respectively. At 10 weeks, total
& free testosterone were 26.6 nmol/l (766.5 ng/dl) & 239 pmol/l,
respectively. This represents a total & free testosterone increase
of 1064% & 362%. Notably, at the 15-day mark, total
testosterone was increased by 708%.
Clearly, the 42% testosterone increase by D-Asp is not in the
same ballpark as the effects from pharmacological doses of
AAS. The glaring lack of human safety & efficacy data on D-
Asp make its use very difficult to justify. Could future research
eventually legitimize D-Asp as the next big supplement
breakthrough? Nothing’s impossible, but right now it’s way too
early to make any confident speculations. At the risk of eating
my words, I wouldn’t bet on it.
References
1. D'Aniello A. D-Aspartic acid: an endogenous amino acid with
an important neuroendocrine role. Brain Res Rev. 2007
Feb;53(2):215-34. Epub 2006 Nov 21.. [Pubmed]
2. D'Aniello S, et al. D-Aspartic acid is a novel endogenous
neurotransmitter. FASEB J. 2011 Mar;25(3):1014-27. Epub
2010 Dec 16. [Pubmed]
3. Topo E, et al. The role and molecular mechanism of D-aspartic
acid in the release and synthesis of LH and testosterone in
humans and rats. Reprod Biol Endocrinol. 2009 Oct 27;7:120.
[Pubmed]
4. US National Library of Medicine/National Institutes of Health.
Testosterone. Updated Jan 21, 2010. [MedlinePlus]
5. Multiply any value expressed in ng/dl by 0.0347 if you need to
convert the value to nmol/l. [JAMA]
6. Bhasin S, et al. The effects of supraphysiologic doses of
testosterone on muscle size and strength in normal men. N Engl
J Med. 1996 Jul 4;335(1):1-7. [Pubmed]
7. Bhasin S, et al. Testosterone replacement increases fat-free
mass and muscle size in hypogonadal men. J Clin Endocrinol
Metab. 1997 Feb;82(2):407-13. [Pubmed]
[Back to Contents] Page 14

Chi L. Chiu comments on my review of Bray et al.
Editor’s note: Special thanks is due to Chi L. Chiu for offering
the following feedback. Anyone who wants to discuss this further
may contact him through his Facebook page.
__________________________________________________
Hi Alan,
Some quick thoughts on the Bray GA 2012 review (I did not
read the publication, just your review).
You mention that the first law of thermodynamics suggests, that
the low protein group should add more fat mass than the medium
and high protein group, which it did not. You suggest that it may
come in time, when the gaining of fat free mass wears off, which
sounds reasonable. I do however, have some additional thoughts
on the matter, that can explain this result:
1. I noticed that the amount of carbs are consistent in all groups.
The body tends to burn of glucose first and prefers storing fat as
fat, as demonstrated by several isotope studies (Schwarz JM
2002, Strawford A 2004). This makes perfect sense, because
why would you want to waste energy on converting glucose into
fatty acids, when you can burn it off. The medium and high
protein groups in the Bray study have less fat than the low
protein group. So there is less to store as fat, which leaves carbs.
Why didn't the low protein group store the excess carbs as fat?
2. The increase of calories would almost certainly lead to insulin
resistance (Hivert MF 2007, Salans LB 2007). This impairs
glucose uptake and the excess glucose does not turn to fat, but
leaves the body as expensive urine. This happens with diabetes
mellitus (roughly translated as 'pass-through honey-sweetness'),
which was named after this phenomenon. It's the calorie-out part
of the equation, which can lead up to an estimated 400 kcal per
day with diabetes patients. Although I have no idea how insulin
resistant the participants became, I think that it is a reasonable to
assume that glucose leaked that way.
The two factors combined would explain, at least for the most
part, the weight difference between the groups, suggesting that
the first law of thermodynamics is still valid in this experiment.
I was not impressed by the additional amount of fat free mass
(0.75 kg) in the high protein group. You mentioned that they fed
3 g per kg bodyweight, which is more than what is used in
normal practice. The authors applied the gold standard for
energy measurements with a metabolic chamber and doubly
labeled water. They did not hold the same standard for body
composition with the DXA. The gold standard is the four-
compartment (4C) method which is more expensive and more of
a hassle. To assess the claim, we need to know the deviation of
the DXA from a 4C measurement. This has been tested several
times, and although the DXA is quite accurate, it does get
thrown off a bit with body mass changes, which has to do with
the fluctuation of hydration. Differences of 1 or 2 kg between 4C
and DXA measurements are the rule, not the exception (Prior
Alan Aragon’s Research Review – February 2012
BM 1997, Swe Myint K 2010). An additional increase of 0.75
kg fat free mass, due to more protein, can therefore not be
concluded by this experiment.
References
Bray GA (2012), Smith SR, de Jonge L. Effect of dietary protein
content on weight gain, energy expenditure, and body
composition during overeating: a randomized controlled trial.
JAMA. 2012 Jan 4;307(1):47-55. [Pubmed]
Hivert MF (2007), Langlois MF, Carpentier AC. The entero-
insular axis and adipose tissue-related factors in the prediction of
weight gain in humans. Int J Obes (Lond). 2007 May;31(5):731-
42. Epub 2006 Nov 28. Review. [Pubmed]
Prior BM (1997), Cureton KJ, Modlesky CM, Evans EM,
Sloniger MA, Saunders M, Lewis RD. In vivo validation of
whole body composition estimates from dual-energy X-ray
absorptiometry. J Appl Physiol. 1997 Aug;83(2):623-30.
[Pubmed]
Salans LB (1968), Knittle JL, Hirsch J. The role of adipose cell
size and adipose tissue insulin sensitivity in the carbohydrate
intolerance of human obesity. J Clin Invest. 1968 Jan;47(1):153-
65. [Pubmed]
Soenen S (2010), Westerterp-Plantenga MS. Changes in body fat
percentage during body weight stable conditions of increased
daily protein intake vs. control. Physiol Behav. 2010 Dec
2;101(5):635-8. Epub 2010 Sep 29. [Pubmed]
Schwarz JM (2002), Mulligan K, Lee J, et al. Effects of
recombinant human growth hormone on hepatic lipid and
carbohydrate metabolism in HIV-infected patients with fat
accumulation. J Clin Endocrinol Metab. 2002 Feb;87(2):942.
[Pubmed]
Strawford A (2004), Antelo F, Christiansen M, Hellerstein MK.
Adipose tissue triglyceride turnover, de novo lipogenesis, and
cell proliferation in humans measured with 2H2O. Am J Physiol
Endocrinol Metab. 2004 Apr;286(4):E577-88. Epub 2003 Nov 4.
[Pubmed]
Swe Myint K (2010), Napolitano A, Miller SR, et al.
Quantitative magnetic resonance (QMR) for longitudinal
evaluation of body composition changes with two dietary
regimens. Obesity (Silver Spring). 2010 Feb;18(2):391-6. Epub
2009 Aug 20. [Pubmed]
This clip is genuinely hilarious; ego training at its finest. At least
the guys in that video seem to be only about 75% serious. Credit
goes to Matt Perryman for tipping me off to that precious
footage.
If you have any questions, comments, suggestions, bones of
contention, cheers, jeers, guest articles you’d like to submit, or
any feedback at all, send it over to aarrsupport@gmail.com.
[Back to Contents] Page 15