Oregon State University

The

Linus Pauling
Institute
FALL-WINTER 2015 RESEARCH NEWSLETTER

From the Director

Diet and Optimum
Health Conference

Balz Frei, Ph.D.
LPI Director and Endowed Chair
Distinguished Professor of Biochemistry
and Biophysics
Joan H. Facey LPI Professor

I

have decided to step down as LPI director and retire
from Oregon State University at the end of June 2016.
By then, I will have served as director of the Linus Pauling
Institute for 19 years—a very long time for any directorof a center or institute. I have had a very productive andhighly rewarding time here at LPI and OSU, and it has
been a true privilege and pleasure to build the Institutefrom its modest beginnings in Weniger Hall to where weare today in the state-of-the-art Linus Pauling Science
Center. I am proud of our new home in LPSC; all theexcellent faculty, students, and staff I have recruited to
LPI; and the top-notch institute we have built together.
When I joined LPI in 1997, I was the only principal
investigator in the Institute; we now have 12 PIs working
in three major areas of research: Healthy Aging,
Cardiometabolic Disease Prevention, and Cancer
Prevention and Intervention. Together, we have published
933 scientific publications over the past 18 years, whichhave been cited by our peers in their publications over
26,000 times—impressive numbers that attest to our prolificscholarly output and the significant impact of our work.
The most highly cited publication from LPI, with almost
800 citations, is a review article on “Tea catechins andpolyphenols: Health effects, metabolism, and antioxidant
functions,” co-authored by the late Dr. Jane Higdon andmyself. Other highly cited articles focus on the antioxidant
functions and health benefits of vitamins C and E; theroles of oxidative stress and inflammation in aging andneurodegeneration, in particular Lou Gehrig’s disease;the mechanisms by which lipoic acid enhances energy
metabolism and reverses many of the age-related declines
in mitochondrial and cellular functions; the mechanisms
and role of phytochemicals (plant chemicals that may

continued on page 2

Stephen Lawson
Administrative Officer

L

PI convened our eighth Diet and Optimum HealthConference, co-sponsored by the Oxygen Club of
California, on the OSU campus in Corvallis from September
9th to 12th. The conference featured 37 speakers fromaround the world and was organized into three main sessions:• Vitamin E: A Critical Nutrient for Development

and Health• Extending Healthspan: Basic Concepts, Underlying

Mechanisms, and the Role of Diet and Lifestyle• Celebrating the Life and Work of George Bailey

Sixty posters depicting experimental projects were displayed
beginning Thursday evening. Ten of these were selected forconsideration for Young Investigator Awards. Additionally,
ten submitted abstracts were chosen for oral presentations.
Saturday morning featured a public session withpresentations by two LPI Principal Investigators. Balz Frei
discussed “LPI’s Rx for Health—or how to meet yourmicronutrient needs,” while Tory Hagen talked about“Adding life to your years and years to your life.”
Videos of their presentations can be viewed at
http://bit.ly/1LiTnwt.
The Linus Pauling Institute Prize for Health Research
was presented posthumously on Friday evening to George
Bailey of Oregon State University for his seminal work in
toxicology and carcinogenesis.

Vitamin E: A Critical Nutrient forDevelopment and Health, chaired by
Maret Traber (LPI)• Maret Traber (LPI) discussed her latest research on therole of vitamin E (alpha-tocopherol) in zebrafish brain

continued on page 2

Continued from cover — Diet and Optimum HealthConferenceContinued from cover — From the Director

development. Embryos from adult zebrafish fed low
dietary levels of alpha-tocopherol exhibit brain and eye
malformations, and fish die five days after birth. This implies
that vitamin E sufficiency may be very important for thedeveloping fetus in pregnant women even before they knowthat they’re pregnant. Mechanistically, it appears thatvitamin E protects an omega-3 fatty acid (docosahexaenoic
acid, DHA) in the brain from peroxidation and depletion.
Low levels of vitamin E also deplete choline in the brain.
DHA and choline play important roles in neurodevelopment
and in the health of adult brains. Many Americans do
not consume the RDA for vitamin E (22.5 IU/day); foodfortification may be one tactic to improve this.

affect health) found in cruciferous vegetables inpreventing colorectal and other cancers; and how
vitamin D enhances immune function and protects
against infectious diseases—to name just a few.
Our research has been supported by gifts fromdonors like many of you, as well as funding from
Oregon State University, industry, the National
Institutes of Health, and other federal agencies.
Over the past 18 years, LPI researchers have brought
over $50 million from NIH and some other extramural research grants and contracts to the Institute.
In addition, thanks mainly to gifts from planned
giving and bequests from our most generous
supporters, LPI’s endowment has more thanquadrupled since the Institute’s inception at OSU.

• Many people in South Asia suffer from multiple
micronutrient deficiencies (vitamin A, folate, iron, zinc,etc.), resulting in increased infections, impaired growthand cognitive development, and an increased risk fordeath in children and adults. Keith West (Johns Hopkins
University) noted that over 50% of children and womenof reproductive age in Nepal and Bangladesh are vitamin
E deficient, although the situation may be improving as
more vegetable oils—a good source of vitamin E—are
consumed. In Bangladesh, pregnant women with vitamin
E deficiency have a risk for miscarriage about twice thatof women not deficient.

One of the accomplishments I am most proud of
is our Micronutrient Information Center—an online
resource of scientifically accurate and peer-reviewed
information regarding the roles of vitamins, minerals,
phytochemicals, and other dietary factors, including
some food and beverages, in preventing disease andpromoting health. This website has become ourpublic face and is very popular among our peers,
health professionals, the media, and the public,
with over 1.5 million users annually from aroundthe world. It has helped the Institute become oneof the most trusted sources of balanced, evidencebased nutrition information in the world.

• Chandan Sen (The Ohio State University) presented

results from his studies on the protection against stroke
damage by low levels of tocotrienol, a member of thevitamin E family found in oat bran, nuts, wheat, palmoil, and corn. However, alpha-tocopherol did not protect
against stroke-induced neuronal damage in animal models.
Tocotrienol has long-lasting effects because it is stored in
fat tissue and released over time. It works by several
mechanisms, including the support of blood vessel formation,
inhibition of pathways of neurodegeneration, inductionof neuronal survival factors, and enhanced blood flow to
stroke-damaged areas. A large clinical trial to test the effectof tocotrienol on patients at high risk for stroke because
they had already had a small stroke began in 2013.

I am also fond of the Healthy Youth Program(HYP) in the Institute. It is our way to serve thelocal community and instill healthy diet and lifestyle
habits into school children and their families.
As we know, it takes a village to raise a child,and the HYP does an excellent job working withlocal schools, health authorities, and community
organizations to teach children life skills that will
put them on the right health trajectory for the restof their lives. By disseminating their successfulprograms through peer-reviewed publications,
online instruction manuals, and social media,
the HYP does its part to help stem the obesity
epidemic in this country.

• About one-third of Americans are afflicted with metabolic

syndrome, a condition that predisposes to diabetes andheart disease. Weight-loss programs to address thissyndrome have largely failed because long-term complianceis poor. Richard Bruno (The Ohio State University)
discussed how metabolic syndrome lowers vitamin E status.
Metabolic syndrome is associated with inflammation andoxidative stress that decrease intestinal absorption andimpair liver metabolism of vitamin E, suggesting thatvitamin E requirements may be higher for people withmetabolic syndrome. Improved vitamin E status may, inturn, protect against the fatty liver disease nonalcoholicsteatohepatitis (NASH), which often occurs with metabolic
syndrome. In related studies, it was found that the amountof dairy fat consumed with vitamin E had little effect on
its absorption.

The University’s Research Office will initiate anational search very soon for a new director, and in
my final column of the LPI Newsletter next Spring,
I will give you an update on the search. Thank you
for your advocacy and support of me personallyand of our research and outreach programs in the
Linus Pauling Institute over the past 19 years—wecouldn’t have done it without you! I think Linus
Pauling would be proud. LPI

FA L L / W I NT ER 2 0 1 5

2• Mark Levine (National Institutes of Health) talked about
the cellular transport of ascorbic acid (vitamin C) anddehydroascorbic acid (DHA, the oxidation product of
ascorbic acid). Ascorbic acid uses two specific transporters
to enter most cells, whereas DHA uses glucose (sugar)transporters. Mouse experiments revealed that vitamin C
gets into red blood cells as DHA, using the glucose transporters, and then gets reduced to ascorbic acid. Red blood
cells deficient in vitamin C and/or vitamin E become veryfragile and rigid and break apart (hemolysis). Since glucose
competes with DHA for entry into red blood cells, diabetics
may have impaired vitamin C status in these cells, leading
to oxidative stress, hemolysis, and hypoxia (lack of oxygen)in the microvasculature. Additionally, in diabetics low levelsof vitamin C in red blood cells may cause the destruction ofvitamin E in those cells by oxidants, leading to hemolysis.

expectancy and healthspan, particularly in Central Asia,
East Asia, and Central and Eastern Europe. Reducing theintake of salt, processed meat, and sugar and increasing
the consumption of fruit, vegetables, and nuts are therecommended interventions.
• Some animal species live very long lives: the naked molerat has a lifespan of about 30 years compared to about
three years for the ordinary rat, and certain species of batsand mollusks are especially long lived. Steven Austad(University of Alabama) suggested that these animals may
provide insight into extending healthspan in people.
Long-lived animals share an ability to very effectively
maintain protein homeostasis—the resistance to proteinoxidation and aggregation and the means to efficiently get
rid of misfolded and damaged proteins. Understanding themechanisms underlying protein homeostasis may help us
to develop dietary strategies to retard senescence. Ordinary
lab mice are not good models to study healthspan because
they differ so much from wild mice, which are leaner, eat
more sparingly, and consume food with nutrient densities
different from the chow fed to lab mice. Several long-term
studies on caloric restriction in primates to extend lifespanmay have had conflicting results because of significant
differences in the basal diets.

Extending Healthspan: Basic Concepts,
Underlying Mechanisms, and Role of Dietand Lifestyle: Session 1
Risk Factors that Limit Healthspan,chaired by Tory Hagen (LPI) and Viviana Pérez (LPI)• While aging is a major risk factor for chronic disease,most attention has been focused on specific diseases rather
than understanding the basic biology of aging. Felipe
Sierra (National Institutes of Health) urged a new researchemphasis on healthspan (geroscience) rather than disease.
Extending lifespan will delay the onset of morbidities, andextending healthspan will compress those morbidities intoa shorter period. There are problems with using standard
rodent models to study disease because they arehomogeneous and don’t truly mimic human disease.
Older people often have co-morbidities from multiple
diseases or conditions that aren’t represented inanimal models. Also, some cancer immunotherapies
that work in young mice are lethal in old mice.
Research has shown that physiological aging ismalleable, unlike chronological aging.

Inflammation, Redox Biology, and Healthy Aging,chaired by Adrian Gombart (LPI) and Kathy Magnusson (LPI)• Miriam Capri (University of Bologna) addressed the age-

related problem of “inflamm-aging,” which is characterized
by sustained low-level inflammation, chronic activation ofcertain immune cells called macrophages, oxidative damage,and a physiological blurring of “self”
and “non-self.” Inflammation isassociated with many age-relateddiseases like atherosclerosis,
diabetes, and cancer (for moredetails, see http://bit.ly/1HVpG9Z).
While nuclear and mitochondrial
DNA affect longevity, the gut
microbiome also plays a role.
The gut microbiome declines indiversity with age and, incentenarians, exhibits alteredproportions of bacterial phyla.
There are over 100 bacterial phyla
in the gut and over 1,000 species; thebiological function of about half ofthese is still unknown. Inflammation
also occurs in centenarians, but theselong-lived people show resilient adaptionthat increases healthspan.

• Mohammad Forouzanfar (University ofWashington) discussed the Global Burden of
Diseases, Injuries, and Risk Factors study, a
global effort involving millions of subjects andover 1,000 scientists in 188 countries thatbegan in 1990. Dietary intake of thesubjects was assessed every five years,
and data on disease incidence,
disability, and mortality werecollected and analyzed.
Deaths attributable topoor diet increased
during the study
period due topopulation growth
and aging. Poor diet(high intakes of salt,
processed meat, andsugar) reduced life

continued on page 4
The naked mole rat has a lifespan of
about 30 years compared to about three
years for the ordinary rat.

3

THE LINUS PAULING INSTITUTE

Continued from page 3 — Diet and Optimum HealthConference• Senescent cells have stopped dividing, but they continue

to be metabolically active and secrete pro-inflammatory
molecules that help in wound healing and injury repair but
are otherwise damaging. Their accumulation in organs andtissues is associated with age-related diseases. Darren Baker(Mayo Clinic) explained that smoking, excessive alcohol
intake, high-fat diets, and other factors induce cellularsenescence. Vitamin E and other antioxidants, as well as
exercise, may be protective. In specially bred mice, thegenetic removal of senescent cells delays the onset ofage-related diseases like cancer and increases lifespan.
• Dean Jones (Emory University) and Helmut Sies

proposed the “redox code”—a set of principles that
underlie the metabolism of oxygen and the way thatorganisms interact with their environment. The relationshipbetween the cumulative exposure to environmental
factors—called the exposome—and the genome largely
predicts health and disease. Oxygen is central to theinterface between the exposome and genome. Animals
evolved in an oxygen-rich environment, and their cells use
oxygen and reactive oxygen species like hydrogen peroxide
as signaling molecules. Aging is associated with a decline
in the adaptability of the “exposure memory systems”
that govern the response to the environment.

Strategies to Maintain Cognitive Function,

chaired by Kathy Magnusson (LPI) and Adrian Gombart (LPI)• Kathy Magnusson (LPI) discussed the cognitive declinethat accompanies aging: worsening memory, attention,
cognitive flexibility, and executive function (decisionmaking, organizing goal-oriented behavior, andprioritization). World and verbal knowledge, as well as
short-term memory and language, are largely preserved inolder age. Rodents share similar declines in cognition withpeople, so they are good models for research. The aging
brain retains some plasticity—studies have shown thatexercise, an enriched environment, and mental training,as well as caloric restriction, dietary polyphenols, andpolyunsaturated fats, can improve cognitive and motor
functions like balance and gait instability in older age.
Although the number of neurons doesn’t decline much withage, they shrink and the number of synapses decreases.
Exercise increases neurogenesis and brain volume, anenriched environment increases neurogenesis, vitamin D
increases certain brain receptors, lipoic acid improves longterm memory, and a Mediterranean diet that emphasizes
polyunsaturated fats and flavonoids helps old brains.

Vitamin D increases certain brain
receptors, lipoic acid improves long-term
memory, and a Mediterranean diet thatemphasizes polyunsaturated fats andflavonoids helps old brains.

• While long-term caloric restriction has been shown inseveral animal species to extend lifespan, its more important
effect may be in extending healthspan by attenuating agerelated cognitive decline and delaying the onset of chronic
diseases. Because meaningful caloric restriction must be
severe (20-50% reduction in calories) and long term to
FA L L / W I NT ER 2 0 1 5

4

manifest benefits, investigators have searched for
substances that mimic caloric restriction without problemsof compliance. Donald Ingram (Louisiana State University)
discussed some of these compounds, including resveratrol(found in peanuts, grapes, and wine) and rapamycin(a naturally occurring antibiotic found in specific soil).
Caloric restriction mimetics typically affect glucose
metabolism, insulin sensitivity, and/or autophagy, theprocess by which cells digest and recycle damaged
molecules. Many caloric restriction mimetics also improve
motor function and cognition in older animals. Proof ofefficacy in people is very difficult to obtain.

primary reason for emergency room visits in older adults.
Falls often result in fractures of the hip, wrist, or spine,with a higher rate among women. Osteoporosis and low
vitamin D status increase the risk for fractures from falls.
Falls are also the major cause of death from injury. Hip
fractures cause enormous disability and result in high rates
of mortality. Vertebral fractures cause chronic, disabling
pain and are associated with a two-fold increase in the riskfor death. Falls are associated with polypharmacy (patientstaking multiple drugs) and impaired vision, cognition, andphysical function, as well as back pain and urinary urge
incontinence at night. Walking for exercise helps lower therisk for falls. A large Swedish study found that older adults
that consumed less than three servings of fruit and vegetables
per day had an increased risk for fall-related hip fractures.

• Andrew Scholey (Swinburne University of Technology,
Australia) focused on the role of nutriceuticals, such as
theanine from tea, curcumin from the spice turmeric, andpolyphenols from cocoa, in maintaining good cognitive
function throughout life. In clinical trials, curcumin
improved working memory and attention and decreased
fatigue after four weeks of supplementation. Cocoa polyphenols improved cognition, blood flow, and neurogenesis
in the brain. Theanine enhanced task-relevant cognitive
functions, and broad micronutrient supplements improved
cognition, mood, sleep, energy, and alertness, possibly bycorrecting insufficiencies. Many of these functions can
now be studied by neuroimaging techniques that elucidate
effects on the brain vasculature and neuronal networks.

• Sean Newsom (Oregon State University) addressed theimpact of exercise on insulin resistance, which is common
among the 90 million Americans that are pre-diabetic.
Obesity and diabetes are closely linked—lipid accumulation
impairs insulin action. Exercise protects against diabetes
and improves glucose metabolism and insulin sensitivity,
but the effect is short-lived. A ten-day period without
exercise increases blood glucose, but dramatic improvements
in insulin sensitivity lasting up to three days can be seen
after one exercise session, regardless of age. Weight loss andexercise lower the incidence of diabetes in men and women.

• Risk factors for
cardiovascular disease(CVD) change with age,
but age itself is not
sufficient to describe thesechanges. Michelle Odden(Oregon State University)
discussed these relationships, with an emphasis
Beginning in 1979,
on the impact of frailty,
Bailey developed trout
which is the loss ofphysiological reserve andas a low-cost modelincreased vulnerability towith high statisticalstress, assessed by gait
power for studies in
speed, fatigue, and strength.
A body mass index (BMI)
carcinogenesis.
greater than 35 is associatedwith an increased risk for
CVD in adults younger than 60. Conversely, in adults over
70, a low BMI is associated with a higher risk for CVD.
In adults aged 65 to 84, a low systolic blood pressure isassociated with lower risk for CVD, whereas a higher
systolic blood pressure is protective in adults 85 andolder. Low blood pressure in the very old and frail may
predispose to falls and injuries, and high blood pressure
may compensate for increased arterial stiffness. Optimal
blood pressure varies according to frailty. Reduced kidney
function is also linked to a higher risk for mortality inoctogenarians.

Celebrating the Life and Work of George Bailey,

chaired by Rod Dashwood (Texas A&M University) andTom Kensler (University of Pittsburgh)• George Bailey, who died in October 2014, had a seminal

research career at Oregon State University, where he retired
as a Principal Investigator in LPI (http://bit.ly/1S00JdQ).
Richard Scanlan (Oregon State University) gave abiographical sketch of Bailey’s career, which began at OSU
in the Sinnhuber Aquatic Research Laboratory (SARL).
SARL gained widespread recognition in the 1960s due to• Lynn Marshall (Oregon Health & Science University)

continued on page 7

talked about risk factors for falls and bone fractures.
Low-impact falls (from standing height or less) are the5

THE LINUS PAULING INSTITUTE

Valencia, Spain

Oxygen Club of California
World Congress
he Oxygen Club of California held its annual world congress,
T“Oxidants and Antioxidants in Biology,” in Valencia, Spain,

Oxidized proteins can aggregate and inhibit the cell’s
proteasomes, which are structures that digest and rid cellsof damaged proteins. Such inhibition may contribute to thedevelopment of Alzheimer’s disease. Oxidized proteins also
impair glucose metabolism in cells, contributing to cellularsenescence.

at the end of June. The large meeting featured 45
presentations on nutrients and medicine, inflammation,
aging and longevity, vascular disease, and neurodegenerative
diseases. Some highlights are summarized below.
• Bharat Aggarwal (University of Texas M. D. Anderson
Cancer Center, Houston) discussed the role of spice-derived
phytochemicals, especially curcumin from turmeric (foundin curry), in cancer prevention. Cancer is associated withchronic inflammation and the activation of a DNA
transcription factor called NF-kB, resulting in cell proliferation,
blood vessel formation, and metastases. In cell culture
studies, curcumin inhibits NF-kB through the productionof reactive oxygen species; this inhibition is abrogated bycellular antioxidants like glutathione.

• Leocadio Rodriguez-Mañas (Getafe University Hospital,
Spain) discussed the problems of frailty and sarcopenia inolder adults. Frailty refers to the increased vulnerability anddecreased physiological reserve of strength in older age.
Frailty is associated with inflammation, oxidative stress,
and other dysfunctions that predispose to disability.
An important deficit of frailty is sarcopenia, which ischaracterized by a decrease in muscle mass and anincrease in fat tissue. In sarcopenia, mitochondria, theorganelles in cells that produce energy, decline in number
and function. An effective treatment for frailty involves
aerobic exercise and resistance and strength training(twice a week for 30-45 minutes) and good nutrition.

• Procyanidins are large molecules in cocoa, apples, grapes,
and peanuts that, although not absorbed by intestinal cells,
are associated with decreased risk for colorectal and gastriccancers. Patricia Oteiza (University of California-Davis)
talked about the ways that procyanidins exert beneficial
biological effects. “Lipid rafts” in cell membranes control
many processes, including cell signaling and “tight junctions”
between cells. Procyanidins prevent the disruption of lipid
membranes and help maintain tight junctions between
cells, thereby limiting the inappropriate exposure to certain
molecules. The interactions between procyanidins and cell
membranes is complex and partly depends on the type oflipids in the membranes. Procyanidins are metabolized by
gut bacteria into smaller compounds that can be absorbed
into the blood stream and exert important biological effects.

• Giuseppe Poli (University of Turin, Italy) addressed therole of oxysterols in brain disease. Oxysterols are oxidation
products of cholesterol that, unlike cholesterol itself, freely
cross the blood-brain barrier. In neuronal cell cultures,
oxysterols have been shown to induce the synthesis ofbeta-amyloid, which accumulates in the brains of
Alzheimer’s patients. Chemical modifications of aprotein called Tau are associated with increasing severityof Alzheimer’s disease. Two main types of oxysterols(24OH and 27OH) have conflicting effects on Tau, but the
speaker speculated that the accumulation of 27OH in thebrain may contribute to disease progression.

• Lipoic acid has many beneficial effects in cell cultures androdents: it inhibits inflammatory molecules in endothelial
cells that line blood vessels and arteries, decreases lipid
synthesis, inhibits atherosclerosis, lowers body weight andtriglycerides, and acts as an antioxidant. Balz Frei (Linus
Pauling Institute) presented the results of a recent study
designed to assess the role of lipoic acid on inflammation,oxidative stress, and body weight in overweight and obese
people. Lipoic acid (600 mg/day) or placebo was given to
64 men and women for 24 weeks. Lipoic acid decreased
biomarkers of lipid oxidation and inflammation and, in thevery obese and in overweight or obese women, lowered
body weight.

• Soy isoflavones have been studied for a possible role incancer prevention, although the evidence to date isinconclusive. José Viña (University of Valencia, Spain)
reported remarkable effects of one soy isoflavone, genistein,
fed to mice used as models for Alzheimer’s disease. The
amount of beta-amyloid, a hallmark of Alzheimer’s disease,in the brain was significantly reduced, and that correlatedwith improved cognition, including learning, memory, andodor discrimination. Clinical trials are needed to confirm
this therapeutic approach in humans.
The 2015 Health Sciences Prize was given to Giuseppe
Poli for his extensive and seminal work on oxidized lipids,
such as hydroxynonenal and oxysterols, and their role inhuman diseases like atherosclerosis, liver fibrosis,
inflammatory bowel disease, and Alzheimer’s disease.

• Aggregated or damaged cellular proteins affect cell health.
Several speakers, including Tilman Grune (German
Institute of Human Nutrition, Potsdam) and Bertrand
Friguet (Sorbonne Universités, Paris), addressed the roleof aggregated or damaged proteins in aging and disease.
FA L L / W I NT ER 2 0 1 5

Additionally, a number of awards, including one
sponsored by the Linus Pauling Institute, were made toyoung investigators. LPI

6

• Ashok Reddy (Oregon Health & Science University)
discussed the implications of Bailey’s large trout studies
on risk assessment in people. Typically, human hazards are
estimated by extrapolating the high doses of carcinogens
used in rodent studies to the low doses that people
encounter in the environment. However, this process has
a lot of unavoidable uncertainty, especially since rodents
have a significant incidence of spontaneous tumors. Troutstudies showed a linear relationship between liver tumors
and low-dose aflatoxin exposure, unlike the non-linear
response to low doses of the widespread environmental
carcinogen dibenzo[a,l]pyrene. While these studies cannot
prove a threshold level of exposure, they are useful topredict a practical threshold and should help the EPA
develop reasonable guidelines.

Continued from page 5 — Diet and Optimum HealthConference
the discovery by Sinnhuber that aflatoxin in moldy cottonseed oil fed to hatchery trout caused liver cancer. Beginningin 1979, Bailey developed trout as a low-cost model withhigh statistical power for studies in carcinogenesis. Bailey
was also renowned as a wine aficionado and for his interestin playing bluegrass music, fly fishing, surfing, and restoring
old trucks.
• Roger Coulombe (Utah State University) discussed theattributes of the trout model to study carcinogenesis.
Trout have a low rate of spontaneous tumors, sensitivity
to many carcinogens, and, after exposure to carcinogens,develop tumors caused by gene mutations relevant tohuman tumors. Trout are also very economical, which, asmentioned above, allows the use of thousands of animals
to achieve high statistical power. For example, Bailey used
42,000 trout to determine the effect of very low-dose
exposures to dibenzo[a,l]pyrene, a ubiquitous environmental
contaminant released by the burning of carbon fuels—
similar to real-world exposures in people.

• Rod Dashwood (Texas A&M University) was a postdoctoral fellow in Bailey’s lab who later worked with him
on chlorophyll studies. Chlorophyll, the green pigment inplants, had previously been shown to be antimutagenic, but
new studies demonstrated its anticancer action. Chlorophyllbinds to aflatoxin, which causes liver cancer, and polycyclicaromatic hydrocarbons in the gut, forming complexes
too big to be absorbed into the blood stream and reachtarget organs like the liver. Indole-3-carbinol from cruciferous
vegetables can block liver cancer caused by aflatoxin
exposure by the same mechanism. Freeze-dried spinach,a terrific source of chlorophyll, inhibited colon tumors inrats treated with heterocyclic amines, or “cooked-meatmutagens,” formed from the high-temperature cooking ofproteinaceous food.

• Glucobrassicin is a compound found in cruciferous
vegetables like broccoli, cabbage, and Brussels sprouts.
When these vegetables are chopped or chewed, an enzyme
converts glucobrassicin into indole-3-carbinol (I3C), which
has been shown in trout studies to inhibit liver cancer
caused by exposure to aflatoxin. In the stomach, I3C isconverted into diindolylmethane (DIM). David Stresser(Corning, Inc.) noted that the protective mechanisms of
DIM are still unknown but may involve cell cycle arrest
and apoptosis (programmed cell death), the disruptionof cell signaling, the induction of enzymes that detoxifycarcinogens, and the inhibition of enzymes that activate
carcinogens. Surprisingly, the timing of the dose isimportant—when I3C is given after the initiation of cancer
by a carcinogen, it may actually promote carcinogenesis.
Therefore, supplementation with I3C may have risks.

• Once experiments had established that chlorophyll and its

derivatives could prevent liver cancer in trout and rodents
exposed to aflatoxin, Bailey was anxious to find out if
people would respond the same way. He and ThomasKensler (University of Pittsburgh) conducted a clinical trial
in Qidong, a region in China where people are unavoidably
exposed to aflatoxin in their food and, consequently, havea very high incidence of liver cancer. The participants were

continued on page 8

OCC World Congress 2016

“REDOX MEDICINE AND NUTRITION”

Convention Center, UC Davis, California

Scientific Topics

May 4-6, 2016

• Redox regulation of cell signaling• Nutrition and the genome-epigenome• Regenerative medicine and wound healing• Bioactives: cognition and aging• Microbiome and gut inflammation

Oxygen Club of California
www.oxyclubcalifornia.org
Department of Nutrition,
University of California, Davis

Organizers: Cesar Fraga, Patricia Oteiza, Helmut Sies
Scientific Advisory Committee: Lester Packer, Maret Traber,
Chandan Sen, Gino Cortopassi, Hagen Schroeter, John Maguire

Co-sponsored by the Linus Pauling Institute7

THE LINUS PAULING INSTITUTE

functions are suboptimal. Deficiencies of iron, zinc, vitamin
A, and iodine are widespread globally, and even a large
percentage of Americans do not meet the RDAs for several
micronutrients. Generally, our intake of B vitamins is
adequate because of food enrichment and fortification, but
the intakes of “shortfall nutrients” like calcium, magnesium,
potassium, and vitamins A, C, D, and E are low. Even a“perfect” diet does not provide adequate amount of vitamins
D and E, potassium, and choline. Most people do not meetthe recommended Dietary Guidelines for Americans for
consumption of whole grains, fruit, vegetables, seafood,and fiber, and exceed the recommended intake of saturated
fats, added sugars, refined grains, and sodium. Weight loss,
the DASH diet, and daily aerobic exercise all reduce bloodpressure more than the reduction of dietarysodium. The Physicians’ Health Study II, along-term, large-scale, randomized trial toassess interventions on disease risk, foundthat a daily multivitamin/mineral significantly
reduced the incidence of total cancers among
male physicians, a presumably well-nourishedgroup. It is confidently estimated that a large
percentage of chronic diseases are preventableby diet and lifestyle.

Continued from page 7 — Diet and Optimum HealthConference
given either a placebo or 100 mg of chlorophyllin (a stablecompound derived from chlorophyll) three times a day justbefore each meal for four months. After three months, urine
was collected and examined for markers of DNA damage
that reasonably predict cancer. The level of such markers in
the urine of people who got the chlorophyllin was 55% lessthan those getting placebo, suggesting that their cancer risk
is lower. Current trials in China utilize broccoli preparations
containing glucoraphanin and sulforaphane to evaluate
their effect on the detoxification of air pollutants.
• Dave Williams (LPI) was Bailey’s

colleague at OSU for many years.
They worked together to establish
the pharmacokinetics (absorption,
metabolism, and excretion) of
aflatoxin and to develop a method to
extract chlorophyll from spinach at
low cost. About 95% of the human
exposure to carcinogenic polycyclicaromatic hydrocarbons (PAHs) comesfrom food. Recent studies have used
accelerator mass spectrometry toestablish the pharmacokinetics of a
PAH called dibenzo[def,p]-chrysenethat will enable the assessment ofvery low-dose exposure to PAHs andpossible micronutrient or dietaryinterventions.

Eighty percentof Americans over
65 have at least one
chronic disease, andmany suffer fromage-related declines
in the quality of life.

• Life expectancy was almost constant until
1900. By WW II, it had doubled in the US
and is 82 years today. As Tory Hagen (LPI)
noted, the increase is due largely to better
sanitation, antibiotics, vaccinations, perinatalcare, and drugs. Once-lethal diseases havenow become chronic diseases that limit
healthspan—that portion of your life when
you are free from debilitating disease. Eighty
percent of Americans over 65 have at least one chronic
disease, and many suffer from age-related declines in thequality of life. Neurodegenerative diseases are common inpeople over 80, and older adults often experience weight
loss, exhaustion, weakness, and slowness due to declines
in cardiac function, nerve conduction, hearing, breathing
capacity, and renal blood flow. Only about 15-25% of thelongevity quotient is genetically determined; the majority is
influenced by the environment, including diet. Five modifiablerisk factors decrease longevity: 1) sedentary lifestyle,leading to frailty; 2) hypertension, leading to kidney andheart failure; 3) obesity, leading to cancer and cognitive
disorders; 4) diabetes, leading to cardiovascular andcognitive declines; and 5) smoking. Many factors areassociated with increased longevity, including marriage,
frequent social contacts, moderate alcohol intake, andmoderate physical activity, which is beneficial for all age
groups. Restricting calories improves health and longevity,
but antioxidants don’t significantly extend lifespan or lower
oxidative damage in old animals. Hundreds of candidates
to extend healthspan are being examined, including dark
chocolate, coffee, red wine constituents, berries, sulfurcontaining compounds in cruciferous vegetables, and caloric
restriction mimetics like rapamycin. Even low levels ofstress may be beneficial by inducing a positive biologicalresponse—a concept known as “hormesis.” LPI• John Groopman (Johns Hopkins
University) gave an overview of cancer and Bailey’s workon aflatoxin and chlorophyll. Although about 50% of
global cancer deaths occur in Asia and only 13% in North
America, most research has focused on cancers relevant
to North America and Europe. From 1991 to 2011, theincidence of cancer was lower than expected, primarily due
to the declining use of tobacco. Liver cancer, which mainly
afflicts those under 50 years old, remains the second
leading cause of global cancer deaths. The dietary
substitution of rice for corn (which is more likely tocontain aflatoxin) beginning in about 1985 correlateswith the dramatic reduction in liver cancer, and Bailey
played a central role in developing and understanding
dietary phytochemical interventions.

The Public Symposium on Saturday morning featured

two of LPI’s Principal Investigators: Balz Frei, who discussed
LPI’s “Rx for Health” (http://lpi.oregonstate.edu/rx-health)and Tory Hagen, who talked about increasing healthspan.
• As Balz Frei (LPI) noted, micronutrients themselves provide
neither energy nor calories but are important in energy
conversion, cell and tissue functions, the immune system,
metabolism, and growth and development. There is adifference between frank deficiencies that can lead todisease and death, and inadequacy in which biological
FA L L / W I NT ER 2 0 1 5

8

The Linus Pauling
Institute Prize for
Health Research
The eighth Linus Pauling Institute Prize for HealthAflatoxin is a carcinogenic substance formed by moldthat grows on improperly stored grains. It’s responsible fora high incidence of liver cancer in regions of China where
people are unavoidably exposed dietarily. Aflatoxin also
causes liver cancer in trout. Dr. Bailey and his colleagues
showed that co-administering chlorophyll or its derivative
chlorophyllin with aflatoxin protected trout from liver cancer.
Chlorophyll binds to aflatoxin in the gut, preventing its
absorption into the blood stream and subsequent delivery
to the liver. The chlorophyll-aflatoxin complex is thenharmlessly excreted. These results led to a clinical trial
conducted by Drs. Bailey and Thomas Kensler in China inwhich people exposed dietarily to aflatoxin took a chlorophyllin supplement or placebo three times a day before
meals for four months. Biomarkers of DNA damage—presumably indicative of cancer risk—were measured inurine after three months and found to be decreased byabout half in those who got the chlorophyllin, suggestingthat their risk for liver cancer was substantially decreased.

Research was presented posthumously to Dr. George
Bailey, Oregon State University distinguished professor
emeritus and former LPI principal investigator. Dr. Bailey’s
widow, Carol, accepted the medal from LPI Director Balz
Frei at the Diet and Optimum Health Conference banquet
on Friday, September 11.

The Prize recognizes innovation and excellence in researchon the roles of micronutrients (vitamins and minerals) andphytochemicals in promoting optimum health and preventing
or treating disease and successful efforts to disseminate
knowledge to enhance public health. As can be seen fromthe summaries of presentations at the Diet and Optimum
Health Conference on pages 5, 7, and 8, Dr. Bailey developed
the trout model to enable large-scale and economical
studies with exceptional statistical power to detect theeffects of low-dose exposure to toxins and carcinogens.
Dr. Bailey earned his Ph.D. in biochemistry from the
University of California-Berkeley and joined Oregon State
University in 1979, initially serving as an assistant professorof Food Science and Technology. He was the director of
OSU’s Marine and Freshwater Biomedical Sciences Center,sponsored by the National Institute of Environmental
Health Sciences, from 1985 to 2002. He then became a
Principal Investigator in LPI and continued to work on
dose-response issues in carcinogenesis, especially focusedon aflatoxin and polycyclic aromatic hydrocarbons, which
are ubiquitous environmental carcinogens.

As Balz Frei noted, “George pioneered the use of rainbow
trout as a model for human cancer, which lead to a newparadigm of assessing cancer risk at doses relevant tohuman exposure. And we are also honoring George forhis pioneering work using dietary approaches and phytochemicals such as chlorophyll(in) and indole-3-carbinol
for cancer chemoprevention, which has the potential todramatically reduce rates of hepatocellular carcinoma, thethird leading cause of cancer mortality worldwide, as wellas colon and other cancers in the US and around the world.”
LPI principal investigator Dave Williams, who worked with
Dr. Bailey for many years, said, “For some reason what reallysticks with me is the memory of his hands. I would look at
those long, almost bony, fingers and marvel at the different
tasks they could perform. Taking apart an old beat-up 1940’s
pickup truck engine and transmission, then restoring it topristine condition; picking the banjo at a level good enoughto play with the Sawtooth Mountain Boys; deftly casting a
fly with a rod so that it would drop right in front of a large
trout; hitting a 140-yard 8 iron to within a few feet from thepin; gripping the poles as he glided down the ski slope; and,of course, writing many impactful scientific publications
and successful NIH grants.” LPI

Working with trout, Dr. Bailey and his colleagues were able
to assess the effect of very low doses of carcinogens, a featnot practical with rodents. Trout are relatively easy andeconomical to maintain, and huge numbers—tens of
thousands—can be used in experiments for high statisticalpower. Their metabolism of and response to carcinogens
is similar to people, so results may be reliably extrapolated.
Because of the trout’s sensitivity to carcinogens, low incidenceof spontaneous tumors, and very high statistical power attainedwith large numbers of fish, Dr. Bailey was able to more
precisely estimate human cancer risk from some carcinogens
at the doses we are exposed to environmentally.

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THE LINUS PAULING INSTITUTE

Selected Recent Publications by LPI Scientists
Summarized by Stephen Lawson, LPI Administrative Officer
LPI scientists in boldface
Abstracts of most of these papers can be found in PubMed (http://www.ncbi.nlm.nih.gov/pubmed)

Healthy Aging Program

dyslipidemias. Our circadian clocks that synchronize
behavior and biology with the day/night cycle also becomeless precise with advanced age. The authors found thatlipoic acid fed for two weeks to old rats remediateddyslipidemias, partly through the normalization of thecircadian clock.

Kerr DC, Zava DT, Piper WT, Saturn SR, Frei B, andGombart AF. (2015) Associations between vitamin D levels
and depressive symptoms in healthy young adult women.
Psychiatry Res. 227:46-51
Vitamin D3 levels in serum were measured twice in 185
healthy, young college undergraduates in the Pacific Northwest
during fall, winter, and spring. Almost half of the subjects
were insufficient in vitamin D (<30 ng/ml), and lower
vitamin D levels significantly predicted clinical depression.

Roberts BR, Lim NKH, McAllum EJ, Donnelly PS, HareDJ, Doble PA, Turner BJ, Price KA, Lim SC, Paterson
BM, Hickey JL, Rhoads TW, Williams JR, Kanninen KM,
Hung LW, Liddell JR, Grubman A, Monty J-F, Llanos RM,
Kramer DR, Mercer JFB, Bush AI, Masters CI, Duce JA,
Li Q-X, Beckman JS, Barnham KJ, White AR, and Crouch
PJ. (2014) Oral treatment with CuII(atsm) increases mutant
SOD1 in vivo but protects motor neurons and improves
the phenotype of a transgenic mouse model of amyotrophic
lateral sclerosis. J. Neuroscience 34:8021-8031

Magnusson KR, Hauck L, Jeffrey BM, Elias V, Humphrey A,
Nath R, Perrone A, and Bermudez LE. (2015) Relationships
between diet-related changes in the gut microbiome andcognitive flexibility. Neuroscience. 300:128-140
The authors fed mice either high-fat, high-sucrose, or
normal chow diets for five weeks and then examined their
cognitive and memory performance. Fecal samples werecollected and assayed for bacteria to find out if the dietssignificantly affected the various bacterial populations in
the gut. Long- and short-term memory were impaired in
the mice fed the high-sucrose diet, and the microbiome(the collection of various populations of bacteria) was
significantly altered by both the high-fat and, especially,
the high-sucrose diets, suggesting that high intakes of sugar
or fat may contribute to impaired cognitive function.

A mouse that mimics amyotrophic lateral sclerosis (ALS)
has been used to study the disease and the mutations to an
antioxidant enzyme, copper/zinc superoxide dismutase, thatare associated with a fraction of human cases. The mutatedenzyme loses its affinity for zinc, and the resultant zincdeficient enzyme generates reactive molecules that are toxic
to motor neurons. Superoxide dismutase that is copper
deficient has also been found in mice that mimic ALS. In
this new experiment, when a copper-containing compound,
CuII(atsm), was fed to the mice, the amount of mutant
superoxide dismutase increased and, surprisingly, resulted
in improved physical function and survival. The authors
found that the extra copper normalized the copper
content in the copper-deficient superoxide dismutase inmotor neurons in the spinal cord, thus stabilizing the
enzyme and promoting its normal function.

Pride H, Yu Z, Sunchu B, Mochnick J, Coles A, Zhang
Y, Buffenstein R, Hornsby PJ, Austad SN, and Pérez VI.
(2015) Long-lived species have improved proteostasis
compared to phylogenetically-related shorter-lived species.
Biochem. Biophys. Res. Commun. 457:669-675
Autophagy is the process by which cells get rid of large
molecular aggregations and damaged organelles. It complements
the cellular ubiquitin/proteasome process that removes
damaged and misfolded proteins. The authors examinedthese processes in cells from related species with very long(naked mole rats, sugar gliders, and little brown bats) or
short (mice, opossums, and evening bats) lifespans. Autophagy
was enhanced in the longer-lived species, as was proteasome
activity, except for bats. The authors concluded that maintaining
proteins in their functional state correlates with longevity.

Zamzow DR, Elias V, Legette LL, Choi J, Stevens JF, andMagnusson KR. (2014) Xanthohumol improved cognitive
flexibility in young mice. Behav. Brain Res. 275:1-10
An age-related increase in palmitoylation, a chemical
modification of proteins, results in cognitive deficits.
The authors examined the effect of xanthohumol, aphytochemical from hops used in beer manufacture, on
protein palmitoylation in young and old mice. Althoughxanthohumol did not affect palmitoylation status, it did
improve cognitive flexibility of young, but not old, mice ona phytoestrogen-deficient diet.

Keith D, Finlay L, Butler J, Gomez L, Smith E, Moreau
R, and Hagen T. (2014) Lipoic acid entrains the hepaticcircadian clock and lipid metabolic proteins that have been
desynchronized with advanced age. Biochem. Biophys. Res.
Commun. 450:324-329

Fok WC, Chen Y, Bokov A, Zhang Y, Salmon AB, Diaz V,
Javors M, Wood WH 3rd, Zhang Y, Becker KG, Pérez VI,
and Richardson A. (2014) Mice fed rapamycin have an
increase in lifespan associated with major changes in theliver transcriptome. PLoS One 9:e83988

Lipid metabolism changes with advancing age; thesechanges are associated with metabolic syndrome and other
FA L L / W I NT ER 2 0 1 5

10

grapes and blueberries—synergistically induced thesynthesis of cathelicidin.

Rapamycin is a naturally occurring antibiotic found in thesoil of Easter Island. It’s often used after transplant surgery
to help prevent organ rejection and functions as a caloric
restriction mimetic—it produces effects in animals similar to
those induced by caloric restriction. In this study, theinvestigators fed rapamycin to four-month-old mice and thenanalyzed gene expression in the liver when the mice werevery old (25 months). Rapamycin increased the lifespan ofboth male and female mice and affected gene expression infemales but only in half of the males. Affected genes include
those involved in mitochondrial function (the source of
cellular energy), protein ubiquitination (the process bywhich damaged proteins are targeted for degradation),and, surprisingly and unlike other interventions that extend
lifespan, a reduction in resistance to oxidative stress.

Cardiometabolic Disease Prevention
Angelo G, Drake VJ, and Frei B. (2015) Efficacy of multivitamin/mineral supplementation to reduce chronic diseaserisk: A critical review of the evidence from observational
studies and randomized controlled trials. Crit. Rev. Food
Sci. Nutr. 55:1968-1991
The authors reviewed the scientific literature for randomized
controlled trials and observational studies on the effect ofmultivitamin/mineral (MVM) supplementation on the risk
for chronic diseases like cardiovascular disease and cancer
and age-related eye diseases like cataract and maculardegeneration. Most studies have reported no significant
effect of MVM supplements on chronic disease risk, witha few exceptions. The Physicians’ Health Study II (PHS II),a study of over 14,000 male physicians followed for over
11 years, found that the use of a daily MVM decreased therisk for total cancers by 8%. Another large French study
found that the intake of a daily supplement containing
antioxidant vitamins and minerals for about 7.5 years was
associated with a 30% reduction in the risk for cancer andall-cause mortality in men but not in women. While the
PHS II found that a daily MVM was associated with a 9%
reduction in the risk for total cataract, there was a 38%
increase in the risk for age-related macular degeneration inmen over 70.

Guo C and Gombart AF. (2014) The antibiotic effectsof vitamin D. Endocr. Metab. Immune Disord. Drug
Targets.14:255-266
Heliotherapy—exposing patients to sunlight—has been
employed for centuries, although an understanding of theunderlying molecular mechanism is just emerging. About ten
years ago, it was found that vitamin D is effective in killing
bacteria by stimulating the synthesis of an antimicrobial
peptide called cathelicidin. Subsequently, this mechanism
was found to explain the role of vitamin D in killing M.
tuberculosis. Epidemiological studies have reported thatvitamin D deficiency is associated with an increased risk for
infections, including tuberculosis, respiratory tract infections,and influenza. Cathelicidins are synthesized in a varietyof cells, including immune cells, where cathelicidins
stimulate their activity. Low serum vitamin D levels
hasten the progression of disease in HIV-infected subjects.
A randomized, double-blind, placebo-controlled study
found that 1,200 IU/day of supplemental vitamin D given
to school children decreased the incidence of the flu. In
another study, high levels of cathelecidin in kidney dialysis
patients predicted a decrease in early mortality. However,
several studies have found that vitamin D supplementation
did not affect cathelicidin levels, suggesting that prior
immune activation may be necessary. In healthy humans,
a positive association between vitamin D and cathelicidin
was observed only at levels of vitamin D less than
32 ng/ml in the blood. Also, the regulation of cathelicidin
occurs only in humans and primates, making animalstudies difficult to conduct.

Jump DB, Depner CM, Tripathy S, and Lytle KA. (2015)
Impact of dietary fat on the development of non-alcoholic
fatty liver disease in Ldlr-/- mice. Proc. Nutr. Soc. 18:1-9
Fatty liver disease (steatosis) is quite common and associatedwith abdominal obesity. Simple non-alcoholic fatty liverdisease (NAFLD) can progress to non-alcoholic steatohepatitis(NASH), which causes liver injury, chronic inflammation,oxidative stress, and fibrosis. The authors developed amouse model that mimics human NASH. When these mice
were fed a Western-type diet high in saturated and transfat, sugar, and cholesterol, they developed NASH.

continued on page 12

Heliotherapy—exposing patients tosunlight—has been employed for
centuries, although an understandingof the underlying molecularmechanism is just emerging.

Guo C, Sinnott B, Niu B, Lowry MB, Fantacone ML,
and Gombart AF. (2014) Synergistic induction of humancathelicidin antimicrobial peptide gene expression by vitamin
D and stilbenoids. Mol. Nutr. Food Res. 58:528-536
The cathelicidin antimicrobial peptide is a small molecule
made in the body that attacks pathogens like bacteria. Its
production can be induced by vitamin D, niacin, curcuminfrom turmeric, and other substances. Using cell cultures, theauthors screened a library of 446 molecules to find novel
candidates that stimulate the synthesis of cathelicidin.
When combined with vitamin D, resveratrol—found ingrapes, peanuts, and wine—and pterostilbene—found in

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THE LINUS PAULING INSTITUTE

of their embryos should they become pregnant while
getting insufficient vitamin E. Additionally, a Chinese study
found that infants with higher vitamin E status had better
cognitive function at two years of age.

Continued from page 11 — Selected Publications
Supplementing their diet with the omega-3 fatty acids docosahexaenoic acid (DHA) and eicosapentaenoic aicd (EPA),
found in fish oil, attenuated inflammation, oxidative stress,
fibrosis, and dyslipidemia. DHA was especially effective.

Murer SB, Aeberli I, Braegger CP, Gittermann M,
Hersberger M, Leonard SW, Taylor AW, Traber MG,
and Zimmermann MB. (2014) Antioxidant supplements
reduced oxidative stress and stabilized liver function tests
but did not reduce inflammation in a randomized
controlled trial in obese children and adolescents.
J. Nutr. 144:193-201

Elie MR, Choi J, Nkrumah-Elie YM, Gonnerman GD,
Stevens JF, and Tanguay RL. (2015) Metabolomic analysis
to define and compare the effects of PAHs and oxygenated
PAHs in developing zebrafish. Environ. Res. 140:502-510
Polycyclic aromatic hydrocarbons (PAHs) are ubiquitousenvironmental pollutants and carcinogens formed fromthe burning of carbon-containing fuels. We are exposed tothem in the air and in food. The authors exposed young
zebrafish, a good experimental model for vertebrate
development, to two PAHS and looked for developmental
defects. After five days of exposure, the zebrafish had
impaired protein synthesis, mitochondrial function,neural and vascular development, and cardiac function.

Forty-four obese children and adolescents (average age of
13 years) were given daily antioxidants (400 IU of vitamin
E, 500 mg of vitamin C, and 50 micrograms of selenium)
or placebo for four months. Various metabolic parameters,
including oxidative stress, inflammation, glucose levels,
lipid profiles, and liver enzymes, were measured in blood
samples at baseline and after four months of supplementation. Antioxidant supplementation increased the levelof antioxidants in blood, decreased oxidative stress, andimproved liver function but did not affect inflammation.

Legette L, Karnpracha C, Reed RL, Choi J, Bobe G,
Christensen JM, Rodriguez-Proteau R, Purnell JQ, andStevens JF. (2014) Human pharmacokinetics of xanthohumol, an antihyperglycemic flavonoid from hops. Mol. Nutr.
Food Res. 58:248-255

Traber MG. (2014) Vitamin E inadequacy in humans:
Causes and consequences. Adv. Nutr. 5:503-513
Vitamin E is a family of eight forms, but the humanrequirements are based on alpha-tocopherol, which is the
only form that can reverse deficiency symptoms. Over 90%
of American men and women do not consume 12 mg ofvitamin E daily, the EAR (estimated average requirement).
It’s difficult to assess the impact of vitamin E inadequacy—circulating levels in blood do not correlate well withdietary intakes because of age-related changes in lipoproteinsthat carry vitamin E. Certain genetic defects and fat
malabsorption problems may result in severe vitamin E
deficiency, characterized by cardiomyopathy and neurological abnormalities, including ataxia. Symptoms of mild
deficiency may be subtle, although studies in zebrafish
suggest that vitamin E is crucial for normal brain andneuronal development. Therefore, vitamin E adequacy isimperative for pregnant women. Vitamin E protects fatty
acids in the brain from oxidation and also protects cognitive
function. Some studies found that patients with Alzheimer’s
disease had lower vitamin E plasma levels and that vitamin
E supplementation slowed the progression of the disease.

Xanthohumol, a phytochemical from hops used in the
manufacture of beer, has been shown in rodents to possess
anti-inflammatory, antihyperglycemic, and antioxidant
functions. Its ability to lower blood glucose and attenuate
lipid dysfunction suggests potential therapeutic benefit intype 2 diabetes and metabolic syndrome. The authors gave
three doses (20, 60, or 180 mg) to 24 healthy men and
24 healthy women and then determined maximum bloodlevels of xanthohumol (45, 67, and 133 micrograms/L,
respectively). The concentration of xanthohumol in the
blood peaked about one hour after ingestion and again
after another three to four hours. About half of thexanthohumol was gone from the blood in 18-20 hours.
No adverse effects of xanthohumol were observed, and theresults of this study show that xanthohumol metabolism issimilar in rodents and humans.
Lebold KM and Traber MG. (2014) Interactions between
alpha-tocopherol, polyunsaturated fatty acids, andlipoxygenases during embryogenesis. Free Radic. Biol.
Med. 66:13-19

Michels AJ and Frei B. (2013) Myths, artifacts, and fatal
flaws: Identifying limitations and opportunities in vitamin
C research. Nutrients 5:5161-5192

In this review article, the authors address the importance ofvitamin E, primarily alpha-tocopeherol, in early development.
Vitamin E protects polyunsaturated fatty acids like arachidonic
acid and docosahexaenoic acid—critical for brain development—from oxidation. Vitamin E deficiency depletes vitaminC, and vitamin C supplementation can regenerate vitamin
E from its oxidized form. Alpha-tocopherol deficiency
in zebrafish embryos resulted in impaired neurologicaldevelopment, anatomical malformations, and increased
mortality. Since the USDA estimated that 96% of American
women do not meet the dietary recommended intake forvitamin E, this could have implications for the development
FA L L / W I NT ER 2 0 1 5

Vitamin C has been a focus of research since its discovery
as the anti-scorbutic factor in 1932, yet, surprisingly,
much about its role in health and disease is still unknown.
Vitamin C is difficult to manage in cell culture studies
because media components like metals cause its oxidation.
Most human cells in culture are scorbutic because vitamin
C is not routinely supplied to them, and permanent cell
culture lines have been grown without vitamin C for many
generations, possibly affecting their metabolism and responses
to experimental interventions. Most animals synthesize
vitamin C, so experimental animal models must be chosen

12with care. In humans, age and genetic polymorphisms canaffect how much vitamin C is absorbed into the bloodstream from the gut. Pharmacokinetic studies of vitamin C
have illuminated some issues of absorption and excretionof vitamin C taken orally or intravenously, but these were
performed in a small number of young, healthy subjects andmay not usefully indicate the relationship between intake
and resultant levels in all organs and tissues, especiallyin older or ill subjects. There may be a poor correlation
between assessment of dietary intake of vitamin C, which
is problematic, and plasma levels, further confounding theinterpretation of human studies.

Cancer Prevention and Intervention
Perera T, Young MR, Zhang Z, Murphy G, Colburn NH,
Lanza E, Hartman TJ, Cross AJ, and Bobe G. (2015)
Identification and monitoring of metabolite markers of dry
bean consumption in parallel human and mouse studies.
Mol. Nutr. Food. Res. 59:795-806
The Polyp Prevention Trial (PPT) was a four-year randomized
trial with over 1,900 men and women who had priorcolorectal adenomas removed and either consumed their usual
diet or ate a low-fat, high-fruit and -vegetable, and high-fiber
diet. Participants who reported increased consumption ofdry beans (e.g., baked, kidney, pinto lima, black, and navybeans) had a reduced risk for advanced adenoma recurrence.
The Legume Inflammation Feeding Experiment (LIFE)
study examined the effect of a diet high in dry beans on
inflammation and insulin sensitivity in healthy men. The
authors identified several bean metabolites in the serum ofparticipants in the PPT and LIFE bean consumption studies
and in mice fed a diet enriched with an extract of navy beansthat may serve as biomarkers of bean consumption. Thesebiomarkers will be useful in quantitative and precise studies
on the prevention of colorectal cancer by beans.
Atwell LL, Hsu A, Wong CP, Stevens JF, Bella D, Yu TW,
Pereira CB, Lohr CV, Christensen JM, Dashwood RH,
Williams DE, Shannon J, and Ho E. (2015) Absorption andchemopreventive targets of sulforaphane in humans following
consumption of broccoli sprouts or a myrosinase-treatedbroccoli sprout extract. Mol. Nutr. Food Res. 59:424-433
Sulforaphane is a phytochemical found in cruciferous
vegetables like broccoli, cabbage, and Brussels sprouts thathas anticancer effects in cell cultures and rodents. In thisstudy, the authors determined the absorption of sulforaphane inpeople. Cruciferous vegetables contain glucoraphanin; when
they are chopped or chewed, an enzyme called myrosinase
is released that converts glucoraphanin to sulforaphane,
which is then absorbed into the blood stream. Twenty
healthy adults aged 19 to 50 were given a myrosinase-treatedbroccoli sprout extract or fresh broccoli sprouts. The levelsof sulforaphane metabolites in the urine and plasma of thesubjects who consumed fresh broccoli sprouts were three tofive times higher than those in subjects who got the broccoli
sprout extract, although putative anticancer mechanisms inboth groups were not significantly affected by the doses used.

Sulforaphane is a phytochemicalfound in cruciferous vegetables likebroccoli, cabbage, and Brussels
sprouts that has anticancer effects incell cultures and rodents.

continued on page 14

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Watson GW, Wickramasekara S, Fang Y, Palomera-Sanchez
Z, Maier CS, Williams DE, Dashwood RH, Pérez VI, andHo E. (2015) Analysis of autophagic flux in response tosulforaphane in metastatic prostate cancer cells. Mol. Nutr.
Food Res. 59:1954-1961

Continued from page 13 — Selected Publications
Wong CP, Rinaldi NA, and Ho E. (2015) Zinc deficiency
enhanced inflammatory response by increasing immune cell
activation and inducing IL6 promoter demethylation. Mol.
Nutr. Food Res. 59:991-999

Autophagy is a process in which cellular components likedamaged organelles or proteins are delivered to the lysosome
for degradation. Autophagy is dysregulated in cancer
cells; stimulating autophagy may have anticancer benefits.
The authors treated metastatic prostate cancer cellswith sulforaphane to assess its effect on autophagy.
At physiologically relevant concentrations, sulforaphane
did not affect autophagy in either acute or prolonged
exposures. At much higher concentrations, sulforaphane
did induce autophagy and subsequent cell death.

Chronic inflammation, an abnormally persistent immune
response, is associated with many age-related diseases,
including arthritis, atherosclerosis, diabetes, and cancer.
In this study, biomarkers of inflammation were increasedin zinc-deficient cells in culture and in old mice with agerelated declines in zinc status, indicating abnormal immune
activation. Methylation, a normal chemical modificationof genes and proteins, was also decreased in zinc-deficient
cells and mice.

Atwell LL, Beaver LM, Shannon J, Williams DE,
Dashwood RH, and Ho E. (2015) Epigenetic regulation bysulforaphane: Opportunities for breast and prostate cancer
chemoprevention. Curr. Pharmacol. Rep. 1:102-111
Sulforaphane, an anticancer phytochemical in cruciferous
vegetables, exerts its effects through several mechanisms,
including genetic and epigenetic processes. Epigenetics refers
to changes in gene expression (protein synthesis) not causedby changes to DNA itself. Sulforaphane works against
prostate cancer cells through epigenetic mechanisms,
including activation of tumor suppressor genes (probably
by the inhibition of histone deacetylases [HDACs]),
inhibition of tumor-promoting genes, induction of apoptosis(programmed cell death), induction of cellular antioxidant
defenses, and a reduction of inflammation. In breast cancer
cells, sulforaphane also inhibited HDACs, enhanced antioxidant defenses, and induced apoptosis. More researchis needed to understand fully the effect of cooking on
sulforaphane stability, the distribution of sulforaphane andits metabolites to various tissues, the effect of individual
variability on sulforaphane metabolism, and the role ofgut bacteria in sulforaphane metabolism.

Rajendran P, Dashwood WM, Li L, Kang Y, Kim E,
Johnson G, Fischer KA, Lohr CV, Williams DE, Ho E,
Yamamoto M, Lieberman DA, and Dashwood RH. (2015)
Nrf2 status affects tumor growth, HDAC3 gene promoter
associations, and the response to sulforaphane in the colon.
Clin. Epigenetics 7:102. doi:10.1186/s13148-015-0132-y

Watson GW, Beaver LM, Willams DE, Dashwood RH,
and Ho E. (2013) Phytochemicals from cruciferous
vegetables, epigenetics, and prostate cancer prevention.
AAPS J. 15:951-961

Nuclear factor erythroid 2 (NF-E2)-related factor 2 (Nrf2)
is a transcription factor that migrates into the cell nucleus
when oxidative stress occurs. Once in the nucleus, Nrf2
stimulates the expression of antioxidant genes. The authors
treated normal mice and mice deficient in Nrf2 with a colon
carcinogen, followed by dietary exposure to sulforaphanefrom cruciferous vegetables. As expected, sulforaphaneinhibited colon tumorigenesis and decreased tumor sizein normal mice, accompanied by inhibition of histone
deacetylase (HDAC) activity and increased synthesis of
a tumor suppressor protein called p16. Surprisingly, theNrf2-deficient mice had fewer tumors, less HDAC activity,and lower p16 levels than normal mice through mechanisms
yet to be determined. In people given sulforaphane as abroccoli sprout extract, HDAC activity decreased and p16
levels increased in peripheral blood mononuclear cells.
FA L L / W I NT ER 2 0 1 5

14

Prostate cancer is very common in the United States,
accounting for about 30% of cancer diagnoses. Epidemiologicalstudies suggest that diet is an important determinant of
prostate cancer risk; men who consume lots of cruciferous
vegetables have less risk. Mechanistic research has focusedon two anticancer phytochemicals in cruciferous vegetables:
sulforaphane and indole-3-carbinol (I3C), both of which
are produced from precursors by the enzyme myrosinase,which is released when the vegetables are chopped or
chewed. Sulforaphane induces phase 2 enzymes, which
detoxify and chemically prepare toxins and carcinogens forelimination; suppresses aberrant cell-signaling pathways incancer cells; induces apoptosis (programmed cell death),and inhibits histone deacetylases (HDACs), which areoveractive in cancer cells. I3C also stimulates detoxification
enzymes, induces apoptosis, inhibits androgen signaling,
and indirectly inhibits HDAC activity. LPI

DEVELOPMENTS
A Lasting Legacy of Nutritional ScienceIn my previous column for the LPI research newsletter,
I asked readers to share their stories about Linus Pauling.
I heard over and over how approachable and charming
he was, as well as what a great sense of humor he had.
His impact on the lives of many people around the worldis immeasurable, and we’re fortunate that he was willing topersevere in his vitamin C research despite skepticism fromthe medical community.
What science has shown us in the years since is whatmany of you know first-hand—Dr. Pauling’s revolutionary
views on the importance of orthomolecular medicine arefar from discredited.
Almost 20 years ago, the Linus Pauling Institute of
Science and Medicine moved to Oregon State University,where it became the Linus Pauling Institute. Institute
Director Balz Frei was hired in 1997, and the first threeprincipal investigators arrived at the Institute in 1998,
following a national search for innovative scientists withexpertise in micronutrients and health.
By 2011, when the Institute moved into the Linus Pauling
Science Center, a state-of-the-art research facility thatbrought together all of the Institute’s principal investigators,
three core areas of research had evolved: Healthy Aging(Healthspan), Cancer Prevention and Intervention, andCardiometabolic Disease Prevention. Outreach programs

like the Micronutrient Information Center, the Healthy
Youth Program, and this newsletter allow the Instituteto share the results of that research with people in thecommunity and around the world.
As we look ahead to the Institute’s 20th anniversary
at OSU next year, the Institute has 12 principal
investigators, four adjunct faculty, and 27 researchstaff and graduate students working on research
projects in orthomolecular medicine. From vitaminand supplement studies to investigating the impact ofphytochemicals and the role of the microbiome, ourresearchers are demonstrating just how critical the rightmolecule at the right concentration at the right time canbe for our health. These molecular studies are enabled
by technological advances in mass spectrometry andother instruments and would not have been possible
years ago.
None of these advances at the Linus Pauling Institutewould have been possible without the support of people
like you. As we get ready to celebrate the Institute’s
20th anniversary at OSU, I hope you will consider
making a gift in honor of Dr. Pauling’s vision and tosupport future advances in orthomolecular medicine.
Keep up with news from the Institute between
newsletters on the Linus Pauling Institute’s blog
(http://blogs.oregonstate.edu/LinusPaulingInstitute) andon Facebook at www.Facebook/LinusPaulingInstitute. LPI

Marlys Amundson
Director of Development
Photo by Jim Carroll Photography

See us on Facebook
facebook.com/linuspaulinginstitute

Visit the LPI Blog
blogs.oregonstate.edu/linuspaulinginstitute15

THE LINUS PAULING INSTITUTE

Linus Pauling Institute

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Look for these informative
articles inside!

Create a lasting legacy at the
Linus Pauling InstitutePage 1 ....... From the Director
Page 1 ....... Diet and Optimum HealthConference 2015

A “life-income gift” guarantees anincome stream for life. Additional benefits
may include:

Page 6 ....... Oxygen Club of California
World Congress• Increase your income

Page 9 .......The Linus Pauling Institute Prize
for Health Research
Page 10 ..... Selected Recent Publications
by LPI Scientists
Page 15 ..... Developments

GIVING to theLinus Pauling InstituteGifts in support of research efforts can be madeat any time. Checks should be payable to OSUFoundation for Linus Pauling Institute. Information
on giving is available through the OSU Foundation,
1-800-354-7281, or by writing to the Institute.

(a portion may even be tax-free)• Receive an immediate charitable deduction• Reduce capital gains taxes(if the gift is funded with appreciated property or securities)• Reduce estate tax   Life-income gifts are simple and flexible. You transfer cash, stocks, realestate, or other property to fund the life-income gift. You (or whomever
you designate) receives the income stream for life or for a set number ofyears. Payments may begin immediately or are deferred to a future date,such as retirement. Ultimately, your life-income gift will benefit the Linus
Pauling Institute. Many life-income donors direct their gift to establish an
endowed fund in their name, creating a lasting legacy.
To learn more about gifts that pay income, please click http://bit.ly/
1PAAblM, or call me at 800-354-7281 for a personalized illustration.
Sincerely,
Jeff Comfort, Vice President, Principal Gifts and Gift Planning
Special thanks to Barbara McVicar for editorial assistance and photographs and to authors of signed articles.