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Oregon State University

The

Linus Pauling
Instituter e s e a r c h

n e w s l e tt e r

From the Director

Healthy Aging

Balz Frei, Ph.D.
LPI Director and Endowed ChairOSU Distinguished Professor of
Biochemistry and Biophysics

An interview with Viviana Perez, Ph.D.
Assistant Professor of Biochemistry
and Biophysics
LPI Principal InvestigatorIt’s been a year since we moved into our new home, theLinus Pauling Science Center at Oregon State University.
We are very pleased to work in such modern, state-of-the-art
facilities, which have transformed the Institute and elevatedit to new heights. The open laboratories and shared corefacilities; the bright, well-designed offices and conference
rooms; and the informal meeting spaces throughout thebuilding have created a stimulating intellectual environment
and facilitated new interactions and collaborations. This
has resulted in new collaborative research projects, joint
publications, and multi-investigator grant applications.

It feels like we have finally arrived––we have achieved
the vision we had when the Institute moved here to OSU
from Palo Alto many years ago. This vision was to havea state-of-the-art research and teaching facility where
top-notch faculty and their students and post-docs,
supported by an efficient administrative team, work
together to discover new ways of solving some of themost pressing health-care problems facing our nation.
We all share a conviction that prevention is the medicineof the 21st century, and that nutrition, micronutrients, andlifestyle are key to improving public health and getting thespiraling health-care costs under control.
The overarching goal of our work in the Institute is tohelp people extend their healthy lifespan (healthspan)––how we can stay healthy and vital up to an old age. Healthin this context is not just defined as the absence of chronic
disease but also the absence of infirmity and deficits ofdaily living as we get older, as well as “a state of complete
physical, mental, and social well-being,” as the World
Health Organization defines it. For example, research in
our Healthy Aging Program is discovering new “age-continued on page 2

Q. You’re from South America. Were you born andraised in Chile?
A. Yes. I was born in Santiago, and I lived there until I
moved to San Antonio, Texas, in 2004.

Q. Where did you attend college?
A. I went to the University of Chile and also did my
Ph.D. there.

Q. In what subject did you earn your Ph.D.?
A. Biomedical science. At that time I was working with

Dr. Sierra investigating the function of one protein thatis overexpressed with age in rats.

Q. What brought you to the United States?
A. Aging research.
Q. Where did you go for that?
A. I came to the University of Texas Health Science
Center at San Antonio.

Q. What attracted you to the Linus Pauling Institute?
A. The diversity, the emphasis on healthy aging andmicronutrients, and the future application to translate
my research to humans.

Q. And you didn’t feel you could do that as
successfully at your institution in Texas?
A. The strength of the Barshop Institute for Longevity

and Aging Studies at the University of Texas, where
I worked, is the study of the basic biology of aging,not on translational research.

Q. Do you continue to collaborate with yourcolleagues there?
A. Yes, I do. I am constantly talking with them, and weare publishing papers together.

continued on page 3~ Designated by the National Institutes of Health as a ~• Center of Excellence for Research on Complementary and Alternative Medicine •Continued from cover — From the Director
essential micronutrients”––compounds like carnitine, lipoic
acid, and coenzyme Q that we synthesize in our bodyand obtain from our diet in sufficient amounts when weare young but have a hard time synthesizing and absorbing
as we get older. To overcome these limitations and assure
optimum functioning of all cells and tissues, we may
have to take supplements of these compounds.
Age also may increase the need for certain vitamins
and nutritionally essential minerals. For example, older
adults often do not get enough vitamin C from their dietand may have lower intestinal absorption of vitamin C
than younger people. The capacity of our skin tosynthesize vitamin D upon sun exposure also declines
with age. Since both vitamins C and D are important forgood immune function, inadequate levels in our bodies
may lead to increased susceptibility to infectious diseases
in older adults. An efficient immune system also is
important for surveillance and elimination of malignant
cells, keeping cancer at bay.
Another interesting connection exists between
vitamin C and carnitine in that vitamin C is required fortwo enzymes in the biosynthetic pathway of carnitine.
Carnitine is required for the transport of fat into mitochondria where it is oxidized (“burned”), releasing energy
required for normal metabolism and good functioning ofour body. Hence, inadequate intake of vitamin C maylead to lack of energy in older adults, and vitamin C or
carnitine supplementation may help overcome this deficit.
Two recent studies related to the topic of healthspancaught my eye. The first one, conducted at the Cooper
Clinic in Dallas, Texas, investigated the relationshipbetween “cardiorespiratory fitness” in middle-aged
adults and the development of chronic diseases later inlife. Treadmill time was measured in over 18,000 healthy
men and women when they were at a median age of 49.
After a median time of 29 years, the prevalence of eightchronic conditions in these individuals was assessed:
congestive heart failure, ischemic heart disease, stroke,
type II diabetes, chronic obstructive pulmonary disease,chronic kidney disease, Alzheimer’s disease, and cancerof the colon or lung, the two most common causes ofcancer deaths in the U.S.
Not surprisingly, theinvestigators found that
individuals with higher bloodpressure, serum totalcholesterol, body mass index,
serum glucose, and smoking
prevalence had a higher riskof developing these chronic
conditions. However,individuals with the highest
level of midlife fitness had thelowest incidence of chronic

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Fall/Winter 2012

conditions and spent the last five years of their lives inmuch better health than people with the lowest level offitness at midlife. The authors concluded that “higher
midlife fitness may be associated with the compressionof morbidity in older age”––in other words, whilelifespan was not extended, healthspan was.
The second study was conducted at the National
Institute on Aging (NIA) and investigated the effect ofcaloric restriction on health and survival in rhesus
monkeys. Restricting caloric intake by 10-40% has beenreported previously to extend lifespan in worms, flies,
rats, and mice. But whether the same approach also can
extend lifespan in primates, including humans, is notclear. In this study at NIA, monkeys of different ages
were fed a diet restricted by 30% in calories andcompared to a non-restricted control group. Interestingly,
the investigators found that caloric restriction did
not extend lifespan. This result is different from thatof another study conducted at the Wisconsin National
Primate Research Center (WNPRC) that found asignificant extension of lifespan in calorically restrictedrhesus monkeys. Nevertheless, the NIA study, like the
WNPRC study, found a lower incidence of cancer
and diabetes in those animals that were calorically
restricted. Measures of “metabolic health and overall
function” were also improved, including lower bodyweight; lower serum triglyceride, total cholesterol, andglucose levels; and less oxidative stress.
In trying to explain the difference in results regarding
lifespan extension between the NIA and WNPRC studies,
the NIA scientists noted that the diets fed to the twogroups of monkeys differed significantly. The WNPRC
monkeys received a diet containing 28% sucrose, compared
to only 4% in the diet given to the monkeys at NIA.
The NIA diet, which has a natural ingredient base, also
contains fish oil and phytochemicals, including flavonoids,and protein derived from wheat, corn, soybean, fish,
and alfalfa. In contrast, the WNPRC diet is a “purified”
diet with no added fish oil or phytochemicals, andprotein from a single source called lactalbumin. Hence,
the authors suggested that the WNPRC monkeys on
caloric restriction lived longer than their non-restricted
counterparts because they got less of a bad diet thatcaused the ad libitum-fed (without restraint) control
monkeys to die prematurely. In contrast, the NIA
monkeys got a healthier diet and lived longer, and underthese circumstances, caloric restriction did not extend
lifespan. Dr. Viviana Perez further discusses caloric
restriction in her interview on page four of this newsletter.
Bottom line: maintaining a healthy body weight isimportant, but you don’t have to starve yourself to getthere. A much better way is to be physically active andmaintain a high level of fitness throughout life. And dietcomposition matters for good health. None of this will
help you live longer (except for the morbidly obese),but chances are you will live a much healthier andmore enjoyable, active life up to an old age. LPI

Continued from cover — Interview with Viviana Perez

Q. How do you like LPI’s new building?
A. It’s beautiful. I like the big windows, and the laboratories
are open, which allows you to interact with people inother laboratories. It’s a very nice building.

Q. Functionally, do the laboratories have everything
you need to carry out your research?
A. Oh yes, they have everything that we need to do our work.
Q. What got you interested in aging research?
A. When I was in my second year of graduate school, I didn’tknow what I would like to do. And it was at that time
Dr. Sierra moved to Chile. In a seminar, he talked about
aging and why aging is important to study. I realized that
he was right—we care much about diseases, for example,
heart problems and Alzheimer’s disease, but those diseases
are all associated with aging. If we can delay aging, we candelay not only those diseases but almost all chronic diseases.

Q. So an important risk factor for some of thosediseases is advanced age?
A. Exactly.

A. Exactly! The idea is to delay all chronic diseases until
the end and then die. The goal is to add life to youryears, not necessarily years to your life.

Q. Are there any other rodents that live nearly as
long as the naked mole rat?
A. For rodents, it is the exception. But among othersmall mammals, like bats, we also observe differencesin lifespan—some bats live shorter and some live
longer. For example, evening bats live six years, but
short tail bats live around 30 years.

Q. Where is the naked mole rat found, and how
does it live in its natural environment?
A. It’s found in the northeast of south Africa, around

Kenya. They live underground where the temperature isaround 29-30 degrees Celsius (84-85 degrees
Fahrenheit), with 100% humidity.

Q. Since it lives underground, does its environment
have elevated carbon dioxide levels?
A. Yes, since they are strictly subterranean, but they seemto be insensitive to CO2 levels. Dr. Buffenstein from San
Antonio has data indicating that these animals may haveadapted to their environment by overexpressing Nrf2, atranscription factor that regulates the response to stress.

Q. What do they eat?
A. In the wild, they eat tubers and bulbs. They obtain all
the water that they need through their food.

Q. In your longevity research, you study the naked
mole rat. Why is that rodent especially interestingin research on aging?
A. The naked mole rat is interesting because it has a lifespanmuch longer than expected, considering its body mass.
We expect species that have a similar body mass to have
similar lifespans, but this is not true for naked mole rats.
For example, a mouse with a body mass similar to thenaked mole rats lives, in the best conditions, around three
and a half years. The naked mole rat lives 30 years! We
don’t know the exact mechanisms for that yet, but theoxidative stress hypothesis of aging doesn’t seem to be
applicable to this animal. Interestingly, they live healthily
their whole life until the end, when their physiologicalcondition declines rapidly, and then they die. That is what
we want to achieve in humans—to maximize “healthspan.”

Q. You are a member of the LPI Healthy Aging
Program, which focuses on increasing healthspan.
What exactly is healthspan?
A. Healthspan is the period of your lifespan when you areproductive and without chronic diseases and conditions
that limit your daily living. That is what the Healthy
Aging Program in LPI wants to achieve—an increase inhealthspan. In other words, we would like to compress
morbidity so most of your life can be disease-free and fullof productive vitality.

Q. If you compress morbidity, it would then occur
towards the end of your lifespan instead ofbeing chronic.

Q. Do they get enough calcium for skeletal
development from these tubers? They presumably
don’t need vitamin D.
A. Yes, their vitamin D levels are very low because they
are subterranean. However, when their calcium status
was examined, they looked okay. They are notdeficient in calcium, and calcium metabolism appears
to be independent of vitamin D in these creatures.

Q. You found that the proteasome may play a rolein the naked mole rat’s longevity. What is theproteasome and how does it affect aging?
A. The proteasome is a mechanism or pathway in cells that
degrades abnormal proteins. For example, if you havea damaged protein that got misfolded or aggregatedas a result of an oxidative modification, and it cannot
be repaired, the ubiquitin proteasome system will get
rid of it. If the protein is not in good shape, it will be

continued on page 4

The Linus Pauling Institute3

Continued from page 3 — Interview with Viviana Perezlabeled with ubiquitin, which then will be recognizedby the proteasome, which degrades it, releasing theamino acids that then can be used to make new proteins.

Q. How does the proteasome work in the naked
mole rat?
A. We found that proteasome activity is 30% higher in

the naked mole rat compared to mice. We alsomeasured the ubiquitin protein level, which is anindirect measure of proteasome activity. Incredibly,
the level of ubiquitinated proteins in the naked molerat didn’t change with age and remained very low,
unlike in mice, which is low when young but increases
dramatically with age.

Q. How relevant is that to humans?
A. In studies with human skin fibroblasts, researchers

found that proteasome activity declines with age.
Several publications have shown that proteasome
activity in different species declines with age, so adecline in proteasome activity is associated with theaging process in general.

Q. Why would protein stability be beneficial?
A. Proteins actually carry out all the functions in cells.

For example, most enzymes are proteins, and if they
are not working well, many physiological functions will
decline or fail. If the protein gets damaged a little but
is still working, the cell is not going to spend energy torepair this damage because it’s working adequately. But
if the protein is not functional or starts aggregating, it’sgoing to be degraded because it will cause more damage
to the overall cellular function.

Q. What damages proteins?
A. Oxidative stress is one of the major causes of damage
to proteins.

Q. Is there any method for improving proteasome
activity?
A. Dietary restriction increases proteasome activity. Other

strategies have been tried in animals, tissues, or cells
that increase proteasome activity, but you don’t know
if they will be safe for humans. There is anothermechanism called autophagy that also degrades proteins.
It’s more versatile than the proteasome. Autophagy not
only takes care of damaged proteins but damaged cellorganelles as well. This seems more important because,
for example, when the mitochondria get old, we need toreplace them, and autophagy will accomplish this. It may
be better to find ways to improve autophagy rather thanproteasome activity in humans.

Q. You’ve done several studies on thioredoxin,
a sulfur-containing antioxidant enzyme, in mice.
Does thioredoxin affect lifespan in mice?
A. Yes. We used a transgenic mouse model that has
increased levels of thioredoxin. We found thatthioredoxin was one of the few enzymes that increased
median lifespan in mice. We didn’t find a significant

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Fall/Winter 2012

increase in maximum lifespan, but the median lifespan
was extended, meaning that morbidity was compressed
and the healthy part of the aging curve was extended.

Q. The free radical or oxidative stress hypothesis ofaging postulates that free radicals causingoxidative stress damage biomolecules like DNA
and proteins, and this damage accumulates withtime, causing age-related disease and leading todeath. How well do you think this hypothesis
holds up in light of all the accumulated evidence?
A. During my post-doctoral studies in San Antonio, Texas,

I spent most of my time trying to test the oxidative stresstheory of aging. From these studies we can say thatoxidative stress plays a role mostly in age-related diseases
rather than in longevity itself. This was a very important
observation. Therefore, based on these and other results,
it is necessary to modify this hypothesis. Why? If oxidative
stress is really the key for aging, then if you increase
antioxidant enzymes that are going to get rid of oxidants
like hydrogen peroxide and superoxide, then that should
translate into a longer lifespan, but it doesn’t. Even though
there is less damage to macromolecules like DNA,
proteins, and lipids, animals don’t live longer if weincrease the antioxidant status. So that is telling us thatfor lifespan itself, oxidative stress is not the problem.
But feeding mice a high-fat, unhealthy diet will increase
the rate of metabolic disease, and mice deficient inantioxidant enzymes will develop disease earlier thanwild-type mice. Transgenic mice that have elevatedlevels of antioxidant enzymes will be more protected.
So antioxidants can help protect against some of the
age-related diseases, even if they don’t increase lifespan.

Q. You are also interested in the possible link
between caloric restriction and lifespan.
What is caloric restriction?
A. Caloric restriction is a paradigm that was proposed along time ago, around 1930 or so. It aims to decrease
the amount of calories that an animal consumes, but it
is not malnutrition—there are sufficient vitamins andminerals, so it is only the amount of calories that isdecreased. This has been well studied in mice and rats
and is so far the only paradigm that will increase
lifespan and healthspan in several species.

Q. Why would caloric restriction affect lifespan?
A. Nobody knows the specific mechanism, but we knowthat dietary restriction is an intervention that affects
several parameters of aging. For example, dietary
restriction decreases oxidative stress and oxidative
damage, improves mitochondrial function, increases
proteasome activity and autophagy, and improvesprotein homeostasis. Dietary restriction also improves
several physiological parameters, such as insulin
sensitivity. In other words, the whole system works better.

Q. In a controlled environment, it is very easy tocontrol the diet and limit the amount of calories
that the animals get. Would it be practical for
people to adopt this kind of strategy inanticipation of increased lifespan and/or
protection against age-related disease?

A. No, I think it would be impractical for most people,

mainly because it’s not enjoyable. Also, as you
mentioned, it is an intervention that has been done in aclean environment that minimizes exposure to pathogens.
This is important because caloric or dietary restriction
can impair some responses of the immune system.
For example, dietary restriction protects against some
infections but increases the risk for others. Furthermore,
to get the maximum beneficial effect, you have to
drastically limit calories, which is not acceptable tomost people.

Q. Is that why scientists have looked for chemicalcompounds that might mimic calorie restriction
by engaging similar molecular mechanisms?
A. Yes, that was the goal several years ago when people

started looking for mimetics of dietary restriction.
Studies in invertebrates that can be easily manipulated
genetically have established molecular pathways that arealtered by dietary restriction. Certain chemicals or
supplements, including resveratrol, metformin, andrapamycin, are being cataloged as dietary restriction
mimetics. Of these three, the only one that increased
lifespan and healthspan in mice was rapamycin.

Q. What is rapamycin?
A. Rapamycin is an antibiotic found in the soil of Easter

Island. It’s a natural compound used clinically for organtransplants and for cancer patients because it has
immunosuppressive and antiproliferative effects.

Q. What effect does rapamycin have on mice?
Are there any side effects?
A. Rapamycin extends lifespan not only in mice but also

in invertebrates, such as fruit flies. And in mice, it not
only increases maximum lifespan but also increases
healthspan. The side effects in mice are cataracts andtesticular atrophy.

Q. Does it affect body weight, insulin sensitivity,
or antioxidant status?
A. Studies in mice, rats, and humans have shown that

rapamycin induces glucose intolerance and insulin
resistance. No effects on antioxidant status have been
published, and some studies suggest that long-term
rapamycin use reduces body weight.

Q. Have people been given rapamycin long enoughto observe any adverse side effects like cataracts
observed in rodents?
A. Not that I’m aware of. In humans, rapamycin is only

given to patients with cancer or used in combination
with other drugs for tissue transplantation, and it may
be that the treatment is not long enough to observe
those side effects.

Q. Why is rapamycin used in those particular
medical conditions?
A. For its immunosuppressive and antiproliferative effects.
For example, in organ transplant patients, rapamycin

is used in combination with cyclosporine. Rapamycin
decreases rejection of the new organ, but the mechanism
is not really clear. In cancer, it also reduces theproliferation of cancer cells.

Q. What about effects on gene expression?
Have they been studied in mice?
A. We have done one study on gene expression in mice,and our results showed that rapamycin has somesimilarities to dietary restriction. However, dietary
restriction has a massive effect on gene expression
compared to rapamycin.

Q. You also mentioned resveratrol and metformin.
Resveratrol has attracted a lot of attention
because it’s present in wine and available as asupplement. How effective is resveratrol on someof these parameters compared to rapamycin?
A. Resveratrol mainly works when the animal is under
metabolic stress, so it only extends lifespan andhealthspan when the animal is fed a high-fat diet.
Resveratrol enhances lipid metabolism and othermitochondrial functions in normal mice, but publisheddata show that it doesn’t affect lifespan.

Q. There has been some concern about dosage.
For instance, when you try to extrapolate tohumans, the resveratrol dose that might beneeded to achieve results that you observe inrodents would be impossibly high. Is that true?
A. Yes, this is true. The Intervention Testing Programfrom the National Institute on Aging (NIA) has been
testing the effect of resveratrol on lifespan in mice, andso far, they didn’t find an effect on lifespan even withhigh doses.

Q. What do you think about the recently publishedstudy by scientists at the NIA that reported that
long-term caloric restriction for about 25 years inmonkeys did not affect lifespan?
A. It doesn’t surprise me! Dietary restriction works inorganisms ranging from yeast to mice. The question iswhether it will work in our closest biological relatives,
the primates. A previous study done in monkeys in
Wisconsin had published results different from those
we observed in this study from NIA. Why? Well, I think
there are several possible reasons. For example, the dietused in the Wisconsin study was high in sucrose, which
can be considered a metabolic stress. Also, the genetic
background of the monkeys used in the NIA study isdiverse compared to the homogenous genetic backgroundof monkeys used in the Wisconsin study, and we know
from previous studies done in mice that the benefits of
dietary restriction depend on the genetic backgroundof the animal. For example, it was observed that dietary
restriction had a deleterious effect in some animals. The“ad libitum” control monkeys in the NIA study were notreally fed without restraint; there were some restrictions.
So there are differences, including diet, genetic

continued on page 6

The Linus Pauling Institute5Continued from page 5 — Interview with Viviana Perez
background, and other factors, between these twostudies—and, therefore, they produced different results.

Q. Where is your research heading?
A. I am very involved trying to establish whether rapamycin

is a dietary restriction mimetic. If it is, it has the potential
for translation to humans because it is already usedclinically. If we can identify molecular mechanisms andways to diminish side effects, it would be a good option
for increasing healthspan in humans.

Q. Obesity is associated with increased oxidative
stress and inflammation, both of which arehallmarks of chronic age-related diseases likeheart disease, cancer, and diabetes. Does thecurrent obesity problem in the United Statesconcern you with respect to healthspan?

Briefly. . .
Giana Angelo, Ph.D.
LPI Research Associate
Micronutrient Information Center

A. I agree that obesity is a big concern for our society andas a researcher, I think we can do something to improvehealthspan in this group of people. However, I thinkit’s important to create public consciousness about thisproblem. For example, we have to educate society
about the importance of eating healthful food andexercising.

Q. What do you like to do when you are not in
the lab?
A. I take care of my daughter. She keeps me busy!
My family likes outdoor activities, and we often
ride our bikes. LPI

strongly influence the results of the meta-analysis andrepresent an important methodological limitation.
In the present analysis, the authors pooled data on major
cardiovascular events (all-cause mortality, cardiac death,sudden death, myocardial infarction, and stroke) from 20
RCTs of omega-3 PUFA administration with a combined
total of 68,680 patients.

icosapentaenoic acid (EPA) and docosahexaenoic acid
(DHA) are very long-chain, highly unsaturated fatty
acids derived from alpha-linolenic acid (ALA). ALA is an
essential fatty acid and the parent compound from which
all omega-3 polyunsaturated fatty acids (PUFAs) are made.
Rich dietary sources of ALA are canola oil, flaxseeds, andwalnuts. Though EPA and DHA are generated in the bodyfrom dietary ALA, they can also be obtained directly fromfood (especially oily fish) and supplements. The advantage
of increasing EPA and DHA content in cells and tissues toimprove health and prevent disease is an area of intense
research. In particular, the C20-22 omega-3 fatty acids
EPA and DHA, often referred to as “fish oils,” benefit
cardiovascular health by modulating blood lipid levels,
inflammation, and the function of endothelial cells (cells thatline blood vessels) and cardiomyocytes (heart muscle cells).

An emerging theme from the JAMA study, as well as
several other reviews on this topic, is that omega-3
supplementation appears to be better at prevention(“primary prevention”) than therapy (“secondary
prevention”) of cardiovascular diseases (CVD). Notably,
the vast majority of the pooled RCTs were secondary
prevention trials, meaning that the recruited subjects had
pre-existing CVD or were at increased cardiovascular risk.
Thus, supplementation with omega-3 PUFAs may not
reduce the risk of cardiovascular events in patients witha history of CVD, particularly when used in combination
with drug therapy, such as statins, aspirin, or anti-hypertensive medications. However, observational epidemiologic
studies have consistently found that increased fishconsumption or higher omega-3 PUFA blood levels areassociated with a significantly lower risk of cardiovascular
events in healthy adults.

A recent systematic review and meta-analysis,
published in the Journal of the American Medical
Association (JAMA), concluded that supplementation
with omega-3 PUFAs is not associated with a statisticallysignificant reduction in the risk of major cardiovascular
events. A meta-analysis is a statistical method thatcombines the results from several randomized, clinicalintervention trials (RCTs) that address similar questions.
Combining multiple trials increases the probability ofdetecting smaller group differences if present. However,selection criteria for including or excluding studies can

Beyond cardiovascular health, EPA and DHA are
important for visual and neurological development, exertanti-inflammatory effects, and may slow cognitive declinewith aging (see lpi.oregonstate.edu/infocenter/othernuts/
omega3fa). If you do not regularly consume fish, LPI
recommends a two-gram fish oil supplement several times
per week (see lpi.oregonstate.edu/lpirx2). Consumptionof fish or fish oil may not be suitable for all individuals,
such as vegetarians, vegans, or those with seafood allergies.
Alternative sources of C20-22 omega-3 fatty acids, producedin yeast or algae, are commercially available. LPI

E

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Coenzyme Q10
Victoria J. Drake, Ph.D.
Manager
Micronutrient Information CenterCoenzyme Q10 is a member of the ubiquinone family ofcompounds. Humans can endogenously synthesize
ubiquinones; therefore, coenzyme Q10 is not an essential
nutrient. Coenzyme Q10 is a fat-soluble compound that is
located in cell membranes and lipoproteins. The chemical
structure of coenzyme Q10 allows it to accept and donate
electrons. Coenzyme Q10 is part of the mitochondrial electron transport chain, which is a group of electron carriersin mitochondria that transport electrons to and from eachother in a sequence in order to generate adenosine triphosphate (ATP)—the energy currency of cells. Coenzyme Q10
is also important to help maintain an acid pH in lysosomes,
allowing for normal function of enzymes within that organelle that digest cellular debris and waste. Along with vitamin
E, coenzyme Q10 is an effective antioxidant in cell membranes and lipoproteins.
Although coenzyme Q10 is synthesized by the body, it
can also be obtained from the diet, with the richest sources
being meat, poultry, fish, soybean and canola oils, andnuts. However, the contribution of dietary coenzyme Q10
to tissue levels is not clear. Deficiency symptoms have notbeen reported among healthy individuals, and primarycoenzyme Q10 deficiency is a rare, autosomal recessive
genetic disorder caused by defects in the compound’s biosynthetic pathway. This condition is characterized by low
tissue levels of coenzyme Q10 and, consequently, compromised neuronal and muscular function. Clinical symptoms
in some patients with primary coenzyme Q10 deficiency
improve upon oral supplementation with coenzyme Q10.
Some studies have shown that tissue levels of coenzyme
Q10 decline with age. Decreased plasma levels of coenzyme
Q10 have also been observed in patients with diabetes,
congestive heart failure, and some cancers. Thus, there isconsiderable interest in whether oral supplementation withcoenzyme Q10 can help prevent or treat these and otherconditions. Oral supplements have been shown to increase
coenzyme Q10 levels in plasma and lipoproteins, but it is not
yet clear whether oral supplementation in humans increases
coenzyme Q10 concentrations in other tissues of individuals
with normal endogenous coenzyme Q10 biosynthesis.

There is little evidence on the effect of coenzyme Q10
supplementation on slowing the aging process or
preventing chronic diseases. In rodents, lifelong dietary
supplementation with coenzyme Q10 increased tissue
concentrations of the compound and decreased age-related
DNA damage; however, lifespan was not extended.
To date, research has mainly focused on whether oralcoenzyme Q10 supplementation could treat various
conditions, including genetic mitochondrial disorders,cardiovascular diseases, and neurological diseases.

Mitochondrial encephalomyopathies result from genetic
defects in the mitochondrial electron transport chain.
Symptoms are improved in some patients with mitochondrial encephalomyopathies after coenzyme Q10
supplementation, although patients with genetic defects incoenzyme Q10 biosynthesis experience the most dramatic
improvement. A phase III clinical trial is presently under
way to investigate whether coenzyme Q10 might betherapeutic in patients with other mitochondrial disorders.
Because coenzyme Q10 levels are decreased in the heart
muscle of patients with severe heart failure, several smallintervention trials have examined whether coenzyme Q10
supplementation improves cardiac function in such patients.
A 2006 meta-analysis of ten randomized, controlled trials inheart failure patients found that coenzyme Q10 supplementation (99-200 mg/day for one to six months) resulted in asignificant, 3.7% improvement in left ventricular ejection
fraction, a measure of the heart’s pumping ability. Overall,studies to date are mixed, and large-scale, clinical trials areneeded to determine whether coenzyme Q10 supplementation
has utility as an adjunct to conventional therapy in thetreatment of congestive heart failure. One such large trial
is currently being conducted.
Other studies have looked at whether coenzyme Q10
supplementation prevents the free radical-induced heartdamage that occurs following a heart attack or duringsome types of heart surgery, such as coronary artery bypass
graft (CABG) surgery. Small, placebo-controlled trials havefound that pretreatment with coenzyme Q10 (100-300 mg/
day for 7-14 days before surgery) improves various shortterm outcomes following surgery, such as left ventricularejection fraction, cardiac output, and postoperative
recovery. However, such trials have included relatively
few people and have been limited to CABG surgicalpatients. Studies have also investigated whether coenzyme
Q10 supplementation (60-600 mg/day) in addition toconventional therapy might help patients with angina
pectoris, a condition characterized by chest pain, especiallyduring exercise, due to inadequate oxygen supply to theheart muscle. Most trials have reported improvements inexercise tolerance and electrocardiograms (recordings ofthe electrical activity of the heart, used to diagnose
cardiac arrhythmias, myocardial ischemia, and heart
attack) compared to a placebo. Yet, only two studies todate have found coenzyme Q10 supplementation to reduce
symptom frequency and nitroglycerin consumption.
Hypertension (systolic blood pressure ≥140 mm Hg or
diastolic blood pressure ≥90 mm Hg) affects about onethird of Americans, and nearly as many have prehypertension, defined as a systolic blood pressure of120-139 mm Hg or a diastolic blood pressure of 80-89 mm
Hg. A 2007 meta-analysis of 12 clinical trials in hypertensive patients found that coenzyme Q10 reduced systolicand diastolic blood pressure by 11-17 mm Hg and 8-10 mmcontinued on page 10

The Linus Pauling Institute7

7th international conference onDiet and Optimum HealthMay 15-18, 2013 • Linus Pauling Institute, Oregon State University, Corvallis, OR • USA
Wednesday, May 15
2:00 pm Registration begins at the CH2M Hill Alumni Centeron the Oregon State University campus
3:00

Welcome and opening remarks
Balz Frei, Conference Chair, Linus Pauling
Institute, Oregon State University

OMEGA-3 FATTY ACIDS and FATTY ACIDOXIDATION in CARDIOVASCULAR
and Brain HEALTH
Chairs: Maret Traber and Donald B. Jump
Linus Pauling Institute, Oregon State University

8:30-9:00

Vitamin D requirements for
optimum healthRobert P. Heaney, Creighton University

9:00-9:30

Vitamin D and the brain
Thomas H. J. Burne, The University of
Queensland, Australia

9:30-10:00

Vitamin D and cancer prevention
David Feldman, Stanford University Schoolof Medicine

Coffee/Tea Break

10:00

HEALTH EFFECTS AND MECHANISMS
OF ACTION OF XANTHOHUMOL

3:15-3:45 Health benefits of omega-3 fatty acids
Richard Deckelbaum, Columbia University

Chair: Fred Stevens
Linus Pauling Institute, Oregon State University

3:45-4:15 Omega-3 fatty acids in the modulation of the

10:30-11:00

inflammatory response and neuroprotection
Nicolas G. Bazan, Louisiana State University
Health Sciences Center
4:15-4:45 Stearidonic acid, omega-3 index andcardiovascular diseases

Peter Prehm, Muenster University Hospital,
Germany
11:00-11:30

function and heart failure: From lipotoxicity
to lipoprotection

11:30-12:00

Welcome Reception

Thursday, May 16
6:00 am Organized Walk/Run
7:30-8:30 Breakfast provided

VITAMIN D – HEALTH BENEFITS
BEYOND BONE
Chair: Adrian “Fritz” Gombart
Linus Pauling Institute, Oregon State University

8

Fall/Winter 2012

Xanthohumol for treatment of
metabolic syndrome
Fred Stevens, Linus Pauling Institute,Oregon State Unversity

William C. Stanley, University of Maryland
6:30

Hops, xanthohumol and women’s healthRichard van Breemen, University of Illinois
at Chicago

William S. Harris, Health Diagnostic Laboratory, Inc.
4:45-5:15 Omega-3 fatty acids, cardiac mitochondrial

Xanthohumol inhibits hyaluronan
export and prevents osteoarthritis

12:00-1:30 pm Lunch provided (LaSells Stewart Center)

DIET AND EPIGENETIC IMPACTS ONDISEASE AND AGING
Chairs: Rod Dashwood and Tory Hagen
Linus Pauling Institute, Oregon State University
1:30-2:00

Bioactive food compounds andepigenetic mechanisms in breast cancer
Donato Romagnolo, University of Arizona2:00-2:30

Methylation profiling in breast cancer
prevention

10:30-11:00

Helene McNulty, University of Ulster,
Northern Ireland

Clarissa Gerhäuser, German Cancer Research Center(DKFZ), Germany
2:30-3:00

Modulation of microRNA profiles by
dietary chemopreventive agents
Alberto Izzotti, University of Genoa, Italy

3:00-3:30

Coffee/Tea Break

3:30-4:00

Folic acid deficiency, aortic plaque formation
and DNA methylation in vascular tissue
Susan Duthie, University of Aberdeen, UK

4:00-4:30

11:00-11:30

11:30-12:00

5:00-7:00

Poster Session

12:00-1:05 pm Lunch provided (LaSells Stewart Center)
1:10-4:15

6:00 am
7:30-8:30

Presentation by LPI Director, Balz Frei

4:15

Plenary Lecture by Awardee

Chair: Balz Frei
Linus Pauling Institute, Oregon State University
Chemical biology of pharmacologicalascorbate to treat cancer
Garry Buettner, The University of Iowa
9:00-9:30

Vitamin C supplementation effects on
blood pressure
Edgar R. Miller III, The Johns Hopkins University
School of Medicine

9:30-10:00

Saturday, May 18

PUBLIC SESSION: WHOLE FOOD
APPROACHES TO DISEASE PREVENTION
Co-sponsored by the Moore Family Center for
Whole Grain Foods, Nutrition and Preventive Health,Oregon State University

Chairs: Emily Ho and Balz Frei
Linus Pauling Institute, Oregon State University
9:00-9:30

9:30-10:00

Chair: David E. Williams
Linus Pauling Institute, Oregon State University

Effects of whole grains and nuts on

cardiometabolic syndrome risk factors
Penny Kris-Etherton, Penn State University
10:00-10:30

TBA
Eric Rimm, Harvard School of Public Health

Coffee/Tea BreakMICRONUTRIENTS IN FERTILITY AND
PREGNANCY

Healthy dietary patterns in the preventionof breast cancer and metabolic syndrome

Cynthia A. Thomson, University of Arizona

Vitamin C in brain development andcognitive function
Fiona Harrison, Vanderbilt University

10:00

Reception
Banquet Dinner

6:30
7:30

Organized Walk/Run
Breakfast provided

HEALTH BENEFITS OF VITAMIN C:
BEYOND SCURVY

8:30-9:00

Oral abstracts

AWARD CEREMONY
Linus Pauling Institute Prize for
Health Research(hors d’oeuvres and beverages provided)

Friday, May 17

Micronutrients and the developing brain
Victoria Hall Moran, University of Central
Lancashire, UK

Sang-Woon Choi, Tufts University

TBA

Role of micronutrients in human fertility
Kelton Tremellen, Repromed, Australia

Nutritional influences on epigenetics
and age-related disease

4:30-5:00

Folic acid: Fortification versus
supplementation

10:30-11:00

An integrative, family-based approach
to childhood obesity
David Ludwig, Harvard University

11:00-12:00

Panel discussion/Questions and answers
Preliminary Program (as of October 26, 2012)

For more information about the Conference,
please visit the LPI Web site at
http://lpi.oregonstate.edu/conf2013 orphone the Institute at 541-737-5075

The Linus Pauling Institute9Continued from page 7 —Coenzyme Q10
Hg, respectively. The randomized controlled trials includedin this analysis used doses of 100-120 mg/day. In addition,
several small, randomized, controlled trials have investigated whether coenzyme Q10 might improve vascular endothelial function (blood vessel dilation); the ability of bloodvessels to relax or dilate is compromised in individuals withatherosclerosis, coronary artery disease, or diabetes. A 2011
meta-analysis examined the results of five such trials in thosewith type 2 diabetes mellitus, coronary artery disease, or
endothelial dysfunction. Supplementation with coenzyme
Q10 (150-300 mg/day for four to 12 weeks) was associatedwith a 1.7% increase in flow-dependent endothelialmediated dilation, which may be clinically significant interms of reducing risk for heart attack and stroke.
Because impaired mitochondrial function and oxidative
stress contribute to the pathogenesis of neurodegenerative
diseases, coenzyme Q10 has been studied as a potential
therapy for Parkinson’s disease, Huntington’s disease, andFriedreich’s ataxia. Compared to control subjects, studies
have documented decreased ratios of reduced to oxidized
coenzyme Q10 in platelets (irregularly shaped cell fragmentsthat assist in blood clotting) and higher concentrations ofoxidized coenzyme Q10 in cerebrospinal fluid of patientswith Parkinson’s disease. Additionally, one postmortem
study found lower levels of total coenzyme Q10 in the cortex region of the brain compared to age- and sex-matched
controls; however, no differences were observed in theother brain regions examined, including the substantia
nigra—the site of neuron cell death in the brain stem of
Parkinson’s patients. While results of an early clinical trialof high-dose coenzyme Q10 (1,200 mg/day) were promising,more recent trials have generally not reported beneficial
effects in patients with Parkinson’s disease. In fact, a phase
III trial of coenzyme Q10 supplementation (1,200-2,400 mg/day), in combination with vitamin E, was recently halted
because it was deemed unlikely to be effective in treating
Parkinson’s disease.
Coenzyme Q10 has also been investigated as a potential
treatment for inherited neurodegenerative diseases like
Huntington’s disease and Friedreich’s ataxia. Huntington’sdisease is characterized by selective degeneration of striatal
spiny neurons. Disease symptoms, which include movement
disorders and impaired cognition, typically develop in the
fourth decade of life and progressively deteriorate overtime. Research in animal models of Huntington’s disease

10

Fall/Winter 2012

implicates impaired mitochondrial function and glutamatemediated neurotoxicity in disease pathogenesis. Some studies
in mouse models of the disease have found that dietarycoenzyme Q10 supplementation improves motor performance,
reduces various hallmarks of Huntington’s disease (i.e.,
brain atrophy, ventricular enlargement, striatal neuronalatrophy), and extends survival. Several studies in mice havealso found that co-administration of coenzyme Q10 withremacemide (a drug that antagonizes the neuronal receptor
activated by glutamate) results in greater improvements
in most measured parameters. However, a 30-monthrandomized, placebo-controlled trial in 347 patients withearly Huntington’s disease found that coenzyme Q10(600 mg/day)-racemide co-supplementation did not slow
the functional decline associated with the disease. A phase
III clinical trial administering a much higher dose of coenzyme Q10 (2,400 mg/day) or placebo to Huntington’s disease patients is currently under way.
Coenzyme Q10 has also been investigated as a potential
therapy for Friedreich’s ataxia, an inherited neurodegenerative disease that causes iron accumulation within the
mitochondria, resulting in increased oxidative stress anda decline in mitochondrial function. A pilot study in ten
patients with the condition found that co-supplementation
with coenzyme Q10 (200 mg/day) and vitamin E (2,100 IU/day) led to improvements in cardiac and skeletal muscle
function and also helped to prevent the progressive declines
in neurological function. Another study found decreased
serum levels of coenzyme Q10 and vitamin E in patientswith Friedreich’s ataxia, suggesting that supplementation
with both compounds may provide therapeutic benefit.
Large-scale randomized clinical trials are needed todetermine their efficacy in Friedreich’s ataxia.
Although the utility of coenzyme Q10 supplementation
for prevention or treatment of various diseases requires
further study, even high doses of the compound appear to
be relatively safe. There have been no reports of significant
adverse side effects of oral coenzyme Q10 supplementation
at doses as high as 1,200 mg/day for up to 16 months and
600 mg/day for up to 30 months. Recently, 1,200 mg/day
was proposed as the observed safe level for coenzyme Q10
supplementation. Gastrointestinal symptoms, such as
nausea, diarrhea, appetite suppression, heartburn, andabdominal discomfort, have been reported, but taking
divided doses throughout the day might minimize these
symptoms. The safety of high doses of coenzyme Q10 inpregnancy has not been established. However, use ofcoenzyme Q10 supplements may interfere with certain drugs.
For example, concomitant use of warfarin (Coumadin)and coenzyme Q10 may decrease the anticoagulatory effectof warfarin. Additionally, HMG-CoA reductase inhibitors(statins) used to reduce cholesterol levels may also decrease
biosynthesis of coenzyme Q10, but it is not clear whether
these drugs decrease coenzyme Q10 levels independent ofa reduction in circulating lipids or decrease coenzyme Q10
concentrations in the body’s tissues. For more information,please see the article on coenzyme Q10 in LPI’s Micronutrient
Information Center (lpi.oregonstate.edu/infocenter/othernuts/coq10). LPI

The Oxygen Club of California Meeting:
Oxidants and Antioxidants in Biology
Stephen Lawson, LPI Administrative OfficerT

he Oxygen Club of California (OCC) held its biennial
meeting in Alba, Italy, on June 20-23, co-sponsored,
as usual, by the Linus Pauling Institute. Dr. Maret Traber,
LPI’s Helen P. Rumbel Professor for Micronutrient
Research, is the President of OCC and served as one of theorganizers of the meeting.

induced by oxidized lipids and sulforaphane, a phytochemical in cruciferous vegetables like broccoli. Nrf2 hasbeen found to be inhibited in umbilical cord cells in casesof pre-eclampsia and gestational diabetes, which may
increase the risk of diabetes and heart disease in thoseinfants when adults. Dietary selenium deficiency may resultin increased Nrf2 activity as a compensatory mechanism.

Nutrient-gene Interactions and Epigenetics

The meeting, entitled “Cell Signaling and Nutrient-Gene
Interactions,” featured 32 speakers—including three from
LPI—and covered the following topics: “Nrf-2–driven
Regulation of Antioxidant Defenses,” “Nutrient-gene
Interactions and Epigenetics,” “Novel Roles of Micronutrients,” “Lipid Oxidation and Signaling,” and“Epigenetics, Metabolism, and Aging.” These sessions
were followed by a poster session with over 100 posters
on relevant topics.
In his keynote lecture, Richard Weindruch noted thatstudies in rodents and monkeys have demonstrated thatcaloric restriction leads to an increase in healthspan—the
portion of life free from chronic disease and pervasive
decline in health and vitality. He and colleagues have beeninterested in understanding the molecular mechanisms that
underlie the increase in healthspan in order to discoversubstances that could serve as mimics of caloric restriction
because chronic caloric restriction is extremely difficult forpeople to accept. It’s likely that substances that regulateenergy metabolism may serve as such mimics.

Epigenetics is the study of changes in gene expressionthat don’t change the underlying DNA sequence. Speakers
discussed how chemical modifications of DNA, such as
methylation, or of histones, which are proteins that DNA
wraps around, affect gene expression. Acetylation (additionof acetyl chemical groups) of histones results in genes being
turned on, and the deacetylation of histones causes genes
to be turned off. Histone deacetylase (HDAC) inhibitors,
therefore, keep genes, including tumor suppressor genes,
active. In prostate cancer cells, the phytochemical HDAC
inhibitor, sulforaphane, triggers apoptosis (programmed
cell death). It was also reported that the B vitaminriboflavin lowers blood pressure in heart disease patients
carrying a certain gene, which is prevalent in about 10%of people worldwide. Supplementation with riboflavin
may significantly lower the risk for stroke in such people.

Novel Roles of Micronutrients
New roles for micronutrients and phytochemicals in
health continuously emerge. Pomegranate juice has beenshown to slow the increase in PSA in men after prostatectomy, and cell studies with pomegranate extract suggestthat it is anti-inflammatory. Zinc has an important rolein protecting the developing brain from oxidant injury

continued on page 12

Nrf2–driven Regulation of Antioxidant
Defenses
Nrf2 is a transcription factor (a protein that regulates
gene activity by binding to DNA) typically bound toanother protein called KEAP in the cell’s cytoplasm. Toxins
and reactive oxygen molecules cause Nrf2 to dissociatefrom KEAP and migrate into the nucleus, where it induces
antioxidant and detoxification genes that help protect cells.
However, overexpression of Nrf2 may help cancer cells.
Speakers noted that Nrf2 also attenuates inflammation and is

Helmut Sies presented the Science and Humanity Award to José Viña of the
University of Valencia, Spain (left to right: Maret Traber, Helmut Sies, José Viña)

The Linus Pauling Institute11

Continued from page 11 - Oxygen Club of California
by several mechanisms, and zinc deficiency impairs thesynthesis of the endogenous antioxidant glutathione.
Studies on vitamin E and the risk for heart disease havebeen conflicting, but new evidence suggests that somepatients, particularly diabetics with the haptoglobin 2-2
genotype, may benefit from supplementation. Coenzyme
Q10 may help regulate fat metabolism; supplementation ofobese mice lowers body weight and improves metabolic
parameters. Lipoic acid and its derivatives induce cellularresponses to stress caused by toxins and free radicals.
Lipoic acid has also been found to inhibit atherosclerotic
lesion development in a mouse model of atherosclerosis,
as well as to decrease body weight gain and triglycerides.
Through its effect on transcription factors NF-κB(implicated in inflammation) and Nrf2 (regulator of
detoxification and antioxidant enzymes), lipoic acidreverses the decline in the antioxidant enzyme superoxide
dismutase in the aortas of old rats. Using cigarette smoke
as a source of oxidative stress, scientists showed thatresveratrol found in red wine or curcumin from the
spice turmeric (a major constituent of curry) inhibited
inflammation in mice.

Maret Traber presented the Aging Research Award to Claudio Franceschiof the University of Bologna, Italy (left to right: Lester Packer,
Claudio Franceschi, Maret Traber)

activation and supplementation with nicotinamide, which
is a derivative of the B vitamin, niacin, and the precursor
for the NADH that donates electrons in chemical reactions,improved the survival of neurons and increased the levels of
glutathione, an endogenous antioxidant. Oxidative stressand free radical damage have been associated with aging,but some studies correlate oxidative stress more closelywith frailty in old age caused by declines in skeletal muscle
function and motor neurons. Other studies implicate
changes in sex hormones in age-related frailty. Proteasomes
are cellular structures that degrade oxidized and damaged
proteins. There is an age-related decline in the efficiency
of proteasomes, leading to the aggregation of damaged
proteins associated with the development of age-relateddiseases. LPI

The Lester Packer Young Investigator Award was presented to Insa Ernstof the Christian-Albrechts University, Kiel, Germany(left to right: Maret Traber, Lester Packer, Insa Ernst)

Lipid Oxidation and Signaling
Oxidized fats in the body have a wide array of effects.
While oxidized fats sometimes serve beneficial functions,they are associated mainly with inflammation and damage
to proteins and other biomolecules. Oxidized fats that
accumulate in atherosclerotic lesions may contribute toplaque instability and rupture, which could lead to heart
attacks or strokes. Lipids in immune cells may control
their function and response to inflammatory stimuli.
Oxysterols are formed when cholesterol molecules
become oxidized and are typically involved in pathologicalprocesses like atherosclerosis. In addition to serving as
biomarkers of oxidative stress and inflammation,
oxysterols help regulate lipid metabolism in the liver.

Epigenetics, Metabolism, and Aging
Aging is characterized by numerous declines in function
and increased risk for major diseases. In old mice, Nrf2

12

Fall/Winter 2012

(left to right: John Maguire, Maret Traber, Lester Packer, Angela Mastaloudis)

bequests
LPI is grateful for thethe
we have received frompast year:
following friends thisDr. George B. WhatleyMs. Helen P. Rumbel
Mr. Charles R. Lutz
Ms. Ella C. Brauch

Vitamin C, Lysine, and Lipoprotein(a) in
Atherosclerosis and Angina Pectoris
Stephen Lawson, LPI Administrative Officer

Lipoprotein(a)Although lipoprotein(a) [Lp(a)] was discovered about 50
years ago, surprisingly little is known about its cellular andbiochemical functions. Lp(a) consists of a low-density
lipoprotein (LDL) particle containing an apolipoprotein
B-100 molecule linked to another apoprotein molecule
called apo(a), which determines the functional characteristics
of Lp(a) and largely accounts for its heart disease risk.
Blood concentrations of Lp(a) are genetically determined
and generally highest in blacks and lowest in whites. While
concentrations of Lp(a) are stable in individuals over time,they can vary among people by 1,000-fold. There are many
genetic variations in the size of the apo(a) particle in Lp(a),which accounts for the variable association with cardiovascular pathologies and events.
Many large-scale studies have explored the relationshipbetween Lp(a) and cardiovascular diseases, consistently
finding an association between high Lp(a) levels andatherosclerosis, heart attacks, or stroke; Lp(a) has beenvalidated as a significant, independent risk factor for heartdisease. Despite these significant correlations, the preciserole of Lp(a) in elevating the risk for heart disease andstroke remains speculative. Lp(a) seems to be proinflammatory; it activates NF-κB, a transcription factor
that binds to DNA and stimulates inflammatory processes.
As a consequence, Lp(a) promotes the recruitment of
immune cells called monocytes to the intima, the innermost
layer of blood vessels and arteries facing the blood stream.
Once monocytes have migrated to the intima, they can
become engorged with oxidized LDL—becoming “foam”
cells—and initiate atherosclerotic lesion formation.
Lp(a) also has been found to impair fibrinolysis, theprocess by which fibrin in blood clots is broken down byplasmin, the active form of plasminogen. By interferingwith this process, Lp(a) contributes to thrombosis (bloodclot formation) that could result in a heart attack or
stroke. Clot formation in the arterial wall, called “muralthrombosis,” contributes to the growth of atherosclerotic
lesions; “occlusive thrombosis,” on the other hand, blocks
an artery and triggers a heart attack or stroke. Lp(a) may
also enhance coagulation, which could further contributeto thrombotic events.

The Hypothesis
In 1990, Linus Pauling and a cardiologist colleague
published a paper entitled “Hypothesis: Lipoprotein(a) is asurrogate for ascorbate” in the Proceedings of the National
Academy of Sciences (PNAS) on the putative relationshipbetween vitamin C and Lp(a). They hypothesized that
Lp(a) serves as an evolutionary surrogate or substitute forvitamin C in animals that do not endogenously synthesize
vitamin C, such as humans and monkeys. Indeed, Lp(a) isfound in few species, mainly those that do not synthesize

vitamin C. Since the absence of detectable Lp(a) in most
species—almost all of which synthesize vitamin C—does
not seem to be biologically disadvantageous, it was also
proposed that vitamin C serves as a surrogate for Lp(a).
In a subsequent paper, Pauling noted that our ancestors’
loss of the ability to synthesize vitamin C and theacquisition of Lp(a) synthesis both occurred about
40 million years ago.
As a surrogate for vitamin C and owing to its pro-clotting
functions, Pauling and his colleague suggested that Lp(a)might help repair lesions in the arterial wall caused bymechanical stress, free radicals, and/or sub-optimum
collagen synthesis when vitamin C concentrations are
insufficient. However, atherosclerosis results from thechronic, pathological deposition of Lp(a). A corollary isthat adequate vitamin C status in humans, such as that
achieved by sufficient intake, may help prevent arterial
damage and reduce the need for the deposition of Lp(a)and/or lower its levels, thus preventing the developmentof atherosclerotic plaque. Scurvy caused by vitamin C
deficiency results in capillary fragility and hemorrhage.
Lp(a) may help by inhibiting fibrinolysis, thus preventing
hemorrhage. Pauling and his colleague also suggested thatthe apo(a) part of Lp(a), rich in disulfide groups, may
function as an antioxidant, thereby inhibiting LDL lipidoxidation associated with plaque formation. In this way,
Lp(a) also would serve as a surrogate for the premier watersoluble antioxidant—vitamin C—when its levels are low.
In their first PNAS paper, Pauling and his colleague
proposed that Lp(a) would be present in guinea pigs, a
species unable to synthesize vitamin C. In a subsequent
paper in PNAS in 1990, they showed that, indeed, Lp(a) ispresent in guinea pigs. They further found that vitamin C
deficiency in these animals promoted the developmentof atherosclerotic plaque and that supplemental vitamin C
prevented its development and the accumulation of Lp(a)in the arterial wall. They also suggested that depletion ofvitamin C increases the permeability of the vascular wall,
thereby contributing to the infiltration of Lp(a) and leading
to plaque formation and mural thrombosis. In chronic
vitamin C insufficiency, the prolonged action andaccumulation of Lp(a) may result in the development ofplaque (mural thrombosis) and adverse events like heart
attacks and strokes related to occlusive thrombosis.
The PNAS papers were followed up with a seriesof papers published in the Journal of Orthomolecular
Medicine (JOM) further outlining the putative relationshipbetween vitamin C, Lp(a), heart disease, cancer, diabetes,and other diseases, emphasizing, especially, the role ofchronic vitamin C deficiency in their etiology.

continued on page 14

The Linus Pauling Institute13

Continued from page 13 — Vitamin C

Lowering Lp(a)Unfortunately, neither diet nor exercise has been foundto lower Lp(a) levels. There have been at least threerandomized, controlled trials to evaluate the effect of
supplemental vitamin C on Lp(a) levels. A 1994 study
with 124 healthy men and women aged 17 to 74 detected
no significant effect of 1 or 2 grams per day of vitamin C
for one month on Lp(a) levels. Published in 1995, anotherstudy enrolled 44 patients with coronary heart disease who
received either placebo or 4.5 grams per day of vitamin C
for 12 weeks. The authors reported no statisticallysignificant effect of vitamin C on Lp(a) levels. The last
study, published in 2000, enrolled 101 healthy men andwomen who received either placebo or 1 gram per day ofvitamin C for eight months. Again, the authors reported
no significant effect of vitamin C on Lp(a) levels. These
results conflict with an earlier uncontrolled, small study
published in 1992 with 11 heart disease patients given 9grams per day of vitamin C for 14 weeks. In that study,
vitamin C lowered Lp(a) levels by an average of 27%.
Overall, these studies suggest that supplemental vitamin C
does not lower Lp(a) levels in humans.
However, a few strategies to lower Lp(a) levels have beensuggested by some small studies. One study of 37 heartdisease patients in 2002 found that daily doses of 81 mg ofaspirin for six months lowered Lp(a) levels by an averageof 82% in those subjects with high baseline levels of Lp(a)(>30 mg/dl). Another study in 1998 with 73 hypercholesterolemic patients reported that a daily dose of 2,000 mg
of niacin for 96 weeks lowered Lp(a) levels by an averageof 39%. This dose of niacin is almost 60 times higher than
the tolerable upper intake level—the highest daily intake
likely to pose no risk of adverse effects—of 35 mg/day for
adults. The time-release niacin used in the 1998 study was
generally well tolerated; common side effects included skinflushing and slightly elevated liver enzymes.
Other studies have reported that L-carnitine, a nonprotein amino acid, lowers Lp(a) levels. One study in2000 found that 2 grams per day of L-carnitine lowered
Lp(a) levels by nearly 8% in 36 subjects with high baseline
levels of Lp(a) (40-80 mg/dl). In 2003, a randomized,
placebo-controlled study in 94 diabetic patients found that
Lp(a) levels were lowered by 21% in patients taking 2
grams per day of L-carnitine for six months.

Lysine, Lp(a), and Angina
The amino acids lysine and proline are chemicallyconverted to other forms (hydroxylysine and hydroxyproline, respectively) by vitamin C for collagensynthesis. In the case of chronic vitamin C deficiency,
these conversions cannot take place effectively, andcollagen synthesis is impaired, leading to the loss of thestructural integrity of vascular and other tissues andsubsequent hemorrhage, as previously mentioned. In thissituation, fibrin, by promoting clot formation, may bevaluable in protecting against blood loss. Pauling and his

14

Fall/Winter 2012

colleague speculated that Lp(a) binds excessively to fibrin
in clots, impairing fibrin breakdown by plasmin. They
also speculated that lysine may bind to Lp(a). In this case,lysine might help remove chronically deposited Lp(a)or interfere with its deposition. Thus, lysine may havetherapeutic benefit in atherosclerosis and angina, whichis caused by poor blood flow in arteries compromised byatherosclerosis.
Pauling published three case reports in the early 1990s in
the JOM on the rapid relief from severe, exercise-induced
angina in heart disease patients taking 3-6 grams/day eachof lysine and vitamin C. There are some plausible
explanations as to why vitamin C may have that effect,
mainly related to its beneficial effect on endothelial
function, i.e., its ability to relax blood vessels and improveblood flow via stabilization or increase of tetrahydrobiopterin—an enzyme cofactor that controls nitric oxide
synthesis—but the role of lysine remains obscure. In twoof the three case reports by Pauling, the patients had
been taking large doses of vitamin C, but relief was only
observed after the implementation of lysine. Because therelief from angina was so rapid in the reported cases,
Pauling suggested, as previously mentioned, that lysine—by binding to Lp(a)—may quickly remove Lp(a) fromplaque and also prevent its deposition in developing
plaque. Unfortunately, no controlled clinical trials have yet
been published to validate or refute this strategy to control
angina, and the molecular interactions between lysine andLp(a) aren’t known.
Lysine is found in the diet, and it can be used in the
body as a precursor to synthesize carnitine, an amino acidcritical for mitochondrial energy production in cells. In his
first case report on the amelioration of angina with vitamin
C and lysine, Pauling noted that vitamin C is importantin the hydroxylation reactions that synthesize carnitine
from lysine. A number of studies have found that carnitine
supplementation, as an adjunct to conventional therapy,
is useful in treating heartdisease, including heart
attacks, heart failure,angina, and peripheralarterial disease.

Patents
Several relevant patentapplications were filed
by Pauling and his
collaborator, and threewere awarded: “Use of
ascorbate and tranexamic
acid solution for organ
and blood vessel treatment
prior to transplantation”(1993), “Prevention andtreatment of occlusive
cardiovascular disease
with ascorbate and substances that inhibit the binding oflipoprotein (A)” (1994), and “Therapeutic lysine salt
composition and method of use” (1997).

The first of these described the treatment of organs orarteries used in transplantation with a solution containing
vitamin C and tranexamic acid, both of which were postulated to inhibit the binding of Lp(a) to blood vessel walls,
in order to prevent subsequent atherosclerotic complications. The inventors proposed that this method would havegeneral applications for the prevention and treatment ofheart disease, especially atherosclerosis, and utility forcoronary bypass patients, diabetics, and patients undergoinghemodialysis or organ transplants. The patent awarded in
1994 explained the putative use of vitamin C, tranexamic
acid, lysine, and nicotinic acid (niacin) to reduce plasma
levels of Lp(a) and prevent its binding to the arterial wall,
thereby inhibiting atherosclerosis. The final patent addressed
the use of lysine, vitamin C, beta-carotene, N-acetyl cysteine(a precursor for the synthesis of glutathione, an important
endogenous antioxidant), vitamin E, and nicotinic acid toprevent and treat heart disease. The evidence provided insupport of the patent applications was based on experimentswith guinea pigs and human aortas obtained postmortem.
While animal models of human disease are frequently useful,they do not always predict human outcomes. No data fromhuman clinical trials were presented, so the patent provisions

Developments
Michele Erickson
LPI Director of DevelopmentAll of us associated with the Linus Pauling Institutedeeply appreciate your relationship with LPI andyour deep admiration and fondness for our founder, Dr.
Linus Pauling. It is my privilege to travel the country
to meet many of you and hear your personal stories ofinspiration, connection, and affection for Dr. Pauling.
Often, I am asked, “Is there something meaningful I cando for the Institute that would honor Linus Pauling andhis legacy?” With your encouragement, LPI is launchinga new opportunity to do just that!
After much discussion with people who knew Dr. Pauling,followed his career, and were inspired by his mission torelieve human suffering, LPI identified two priorities forfunding that will bring together your philanthropic intent
and provide a meaningful way to honor Dr. Pauling.
Those conversations helped us to focus on two areasof Dr. Pauling’s expertise that were most meaningful tohim: innovative, cutting-edge research and educating
and mentoring promising new student scientists.
Our vision includes two endowed funds:• The Linus Pauling Endowed ResearchInnovator Fund
• The Linus Pauling Endowed Fellowship Fund

The Linus Pauling Fellowship provides two years of
support for a high-achieving graduate student in LPI.

remained largely speculative. It’s not unusual for patents tobe awarded in order for the inventor to establish priority inadvance of actual proof.

Finally…
We know that vitamin C has value in promoting
cardiovascular health. It not only reduces the risk for heartdisease and stroke but also lowers blood pressure andpromotes vasodilation in patients with atherosclerosis,
thus improving blood flow that may also attenuate angina.
To date, the hypothesized relationship between vitamin C
and Lp(a) has been neither proven nor refuted. It’s also not
yet known if lysine itself, rather than serving simply as aprecursor for carnitine synthesis, plays an important rolein preventing or treating heart disease or angina. And
neither the dose-response relationships nor various
combinations of micronutrients have been adequately
addressed experimentally. Controlled clinical trials areneeded to answer these provocative questions. Lastly, there
is no compelling evidence from controlled trials that any
micronutrient or combination of micronutrients reverses
atherosclerosis in humans, although some may inhibit its
development or retard its progression. LPI
Each new Fellow will be selected and honored publicly at
LPI’s Diet and Optimum Health Conference, with special
recognition of Dr. Pauling and his career as a mentor for
young scientists. This enduring fellowship will be uniquein that it not only supports full tuition and provides
a stipend for the honored student but also provides a
private curriculum experience involving the personal andscientific papers of Dr. Pauling. The curriculum includes
a special emphasis on Dr. Pauling’s interest and work inorthomolecular medicine and nutrition.
The Linus Pauling Endowed Research Innovator
Fund will be awarded to a Principal Investigator in
the Institute on a five-year rotating basis. These funds
will support a faculty member’s research that advances
knowledge in areas that were of interest to Dr. Paulingin the last 25 years of his life. The Linus PaulingResearch Innovator Fund encourages faculty investigators
to pursue cutting-edge research and pilot projects thatneed initial support to establish new research directions.
Such creative and innovative ideas lead to the breakthroughs we need most to advance the mission of theLinus Pauling Institute and the vision of Dr. Pauling.
To learn more about how your investment of funds,
stocks, or a bequest can work with the gifts of other
donors to create an opportunity for perpetual recognitionand honor as a legacy for Dr. Pauling, please contact me
at 800-354-7281 or michele.erickson@oregonstate.edu.
I would love to hear YOUR story of Linus Pauling’s
impact on your life and share ideas on how you canbe part of this profoundly meaningful opportunity tocontinue Dr. Pauling’s mission to reduce suffering andimprove human health for our world. LPI

The Linus Pauling Institute15

Linus Pauling Institute

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Oregon State University
307 Linus Pauling Science Center
Corvallis, Oregon 97331

Portland, OR
Permit No. 1006

phone: 541-737-5075
fax: 541-737-5077
email: lpi@oregonstate.edu
Internet Web site: http://lpi.oregonstate.edu

Gifts in support of research
efforts can be made at any time.
Checks should be payable to
OSU Foundation for
Linus Pauling Institute.
Information on giving isavailable through the
OSU Foundation,
1-800-354-7281, or by
writing to the Institute.

Micronutrient
Research for
Optimum Health
Printed on recyclable paperusing soy inks

The Linus Pauling
Science Center

Photo by Karl Maasdam

GIVING
to theLinus Pauling
Institute

Look for these informative articles inside!
Page 1 ..... From the Director
Page 1 .....Healthy Aging Interview
with Viviana Perez
Page 6 .... Briefly. . .
Page 7 ..... Coenzyme Q10

Page 8 ...... Diet and Optimum Health Conference
Page 11 .... The Oxygen Club of California Meeting
Page 13 .... Vitamin C, Lysine, andLipoprotein(a) in Atherosclerosis
and Angina Pectoris

Page 15 .... Developments

Special thanks to Barbara McVicar for editorial assistance and photographs, authors of signed
articles, and Dick Willoughby for the logo photograph of Linus Pauling.