JANA Vol 9 #1 - American Nutraceutical Association

The Journal of the American Nutraceutical Association 

Vol. 9, No. 1, 2006 www.ana-jana.org 

IN THIS EDITION 

• Nutritional Genomics: From Basic Science to Clinical 

Application 

• The Physiological and Psychological Effects of Animal 

Therapy: Examining the Healing Power of Animals 

• Microdose DNA for the Treatment of Acute and Chronic 

Respiratory Diseases and Otitis Media 

• Phytonutrients May Decrease Obstetric Complications: 

A Retrospective Study 

• Pilot Study: Flaxseed Supplementation Was Effective in 

Lowering Serum Glucose and Triacylglycerol in Glucose 

Intolerant People 

A Peer-Reviewed Journal on Nutraceuticals and Nutrition 

ISSN-1521-4524

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Calcium, Arterial Compliance, Blood Pressure and Anti-oxidant Status in Human Subjects - Mark C. Houston, MD, 

MSc, FACP, Editor-in-Chief, Journal of the America Nutraceutical Association (JANA), Clinical Professor of Medicine, Vanderbilt University School of 

Medicine, ASH Specialist in Clinical Hypertension, Director, Hypertension Institute and Vascular Biology, Clinical Research Staff Physician, 

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and Director of the Prevention Research Center at the Yale University School of Medicine, Dr. Katz is also Associate Director of Nutrition 

Science at the newly established Rudd Center for Food Policy and Obesity at Yale University. 

The Modern NutraGenomics Practice – Guidelines and Updates for the Clinical Practice - Chris Foley, MD. Member, 

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Association 

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Terri Erickson 

ART DIRECTOR 

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Contents – JANA Vol. 9, No.1, 2006 

P E R S P E C T I V E A R T I C L E 

Nutritional Genomics: 

From Basic Science to Clinical Application ..............................1 

Bernd Wollschlaeger, MD, FAAFP 

The Physiological and Psychological Effects of 

Animal Therapy: Examining the Healing Power of Animals .... 7 

Niki Sepsas, Allen Montgomery, RPh 

R E V I E W A R T I C L E 

Microdose DNA for the Treatment of Acute and 

Chronic Respiratory Diseases and Otitis Media ......................13 

Stephen W. Mamber, PhD, John McMichael, PhD 

C L I N I C A L R E S E A R C H 

Phytonutrients May Decrease Obstetric Complications: 

A Retrospective Study ................................................................23 

C. Doug Odom, MD, Suneet P. Chauhan, MD, Everett F. Magann, MD, 

Rick W. Martin, MD, Carl H. Rose, MD, John C. Morrison, MD 

Pilot Study: Flaxseed Supplementation Was Effective in 

Lowering Serum Glucose and Triacylglycerol in 

Glucose Intolerant People ..........................................................28 

Yeong Rhee, PhD, Ardith Brunt, PhD 

To subscribe to JANA phone 800-566-3622, 

outside USA 205-833-1750. 

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To order reprints of articles, or additional copies of JANA, contact: 

Allen Montgomery, RPh 

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Phone 205-980-5710 Fax 205-991-9302 

E-mail: allenm@ana-jana.org 

Website: www.ana-jana.org


Nutritional Genomics: From Basic Science 

to Clinical Application 

During the entire history of medicine humans have 

known that hereditary factors affect health and determine 

the occurrence of certain diseases. Hereditary connections 

between parents and children were apparent, but the mechanisms 

were not readily understood. Only about 150 years ago 

an Austrian monk, Gregor Mendel, developed the fundamental 

principles that would become the modern science of 

genetics. Mendel demonstrated that heritable properties are 

located on discrete units, and are independently inherited. 

These "units" eventually were called genes. Half a century 

ago, Watson and Crick discovered the structure of DNA and 

its function in encoding and transcribing genetic information. 

In 2003, after a monumental effort spanning over 13 

years, the Human Genome Project (HGP) was completed, 

mapping the entire 3.1 gigabase of the DNA sequence. The 

data indicated that the human genome contains 30,000 to 

35,000 genes, 1 but only about half these genes have recognizable 

DNA-sequence patterns that suggest possible functions. 

It is now also apparent that through the mechanism of alter- 

JANA Vol. 9, No. 1, 2006 

Bernd Wollschlaeger, MD, FAAFP* 

Clinical Assistant Professor – Department of Family Medicine & Medicine 

University of Miami School of Medicine 

Associate Editor, Journal of the American Nutraceutical Association 

AMA Delegate and Chair, AMA International Medical Graduates (IMG) Governing Council 

* Correspondence: 

Bernd Wollschlaeger, MD, FAAFP 

Aventura Family Health Center 

16899 NE 15th Ave 

North Miami Beach, FL 33162-2914 

Phone: 305-342-2522 

Email: info@miamihealth.com 

This article is an excerpt from a book written by the author, 

scheduled to be published and released in July 2006 

native splicing2 more than 100,000 proteins can be derived 

from these 30,000 to 35,000 genes, contradicting once prevailing 

dogma that one gene makes one protein. Furthermore, 

we previously assumed that a complex array of molecular signals 

allows specific genes to be "turned on" (expressed) or 

"turned off" in specific tissues and at specific times and that 

not all genes reside on nuclear chromosomes. 3 Several dozen 

genes are located on the mitochondrial chromosome. 

Since ova are rich in mitochondria and sperm are not, 

mitochondrial DNA is usually inherited from the mother. 

An important characteristic of the human genome is that 

most individuals share over 99.9 percent of their DNA 

sequences. 4 However, given the more than 3 billion base 

pairs that constitute the human genome, this also means that 

the DNA sequences of two unrelated humans vary at millions 

of bases. These variants are commonly referred to as 

"single-nucleotide polymorphisms" (SNPs), and these specific 

genotypic variations correlate with specific phenotypic 

variations with often significant impact on health and the 

development of disease. SNPs, also called "snips," can be 

compared to typographical errors occurring in a word (i.e. 

gene) that may change the entire meaning of the sentence 

(i.e. protein) containing that word. For a variation to be 

considered a SNP, it must occur in at least 1% of the population. 

SNPs, which make up about 90% of all human 

genetic variations, occur every 100 to 300 bases along the 

3-billion-base human genome. Two of every three SNPs 

involve the replacement of cytosine (C) with thymine (T). 

1

SNPs can occur in both coding (gene) and noncoding 

regions of the genome. Many SNPs have no effect on cell 

function, but scientists believe others could predispose people 

to disease or influence their response to a drug. Almost 

500 polymorphisms have been identified as having clinical 

importance, and their penetrance affects the expression of 

disease in the affected individual. 

The mapping of the human genome has provided insight 

into the relationship of genes, enzymes, cofactors, substrates, 

and metabolites. We have now acquired the tools to analyze 

the information flow from the genome (genomics) via protein 

translation (proteomics), and from the metabolite network 

and fluxes (metabolomics) to the human phenotype. 

Combined with the understanding of the biochemical pathways 

in human metabolism, we now can also assess especially 

the impact of nutrients on this interrelated complex 

metabolic network spanning from genome to phenotype. 

Epidemiological studies conducted over the last 25 

years that compare dietary patterns (i.e. intake of particular 

food items) between countries of low and high incidence for 

a particular cancer5-6 provided the first clues that diet and 

the consumption of certain nutrients may be a risk factor in 

the development of cancer and chronic diseases through 

interaction with the individual genome. Almost eighty percent 

of colon, breast, and prostate cancer cases and one 

third of all cancer cases may be influenced by diet and associated 

lifestyle factors. Dietary factors that influence the 

expression of cancer genes include carbohydrates, amino 

acids, fatty acids, minerals, and vitamins. Many other nutrients 

that modulate the genetic expression include phytochemicals 

and other components of a plant and fruit-based 

diet including carotenoids, flavonoids, organosulfur compounds, 

isothiocyanates, indoles, monoterpenes, phenolic 

acids, and chlorophyll. 7 

The penetrance or expression of cancer genes is strongly 

influenced by the interaction of both genetic and environmental 

influences. The evidence for this is based on 

studies of human populations as well as from animal experiments 

that model the process of carcinogenesis. 8 Some 

large-scale intervention trials indicated that single nutrients 

cannot explain the beneficial effect of diet on gene expression. 

For example, strong epidemiologic evidence linking 

consumption of carotenoid-rich fruits and vegetables with a 

reduced risk of cancer could not be verified by two other 

large-scale ß-carotene intervention trials. 9-10 These trials were 

conducted with populations of smokers and asbestosexposed 

individuals, revealing that the risk of lung cancer 

increased rather than decreased in the groups supplemented 

with ß-carotene. The trial outcomes mainly reaffirm the 

hypothesis that multiple phytonutrients and not single nutrients 

affect the genetic expression and penetrance that 

results in the development of cancer and other chronic disease. 

Multicenter and projective cohort studies were conducted 

in Europe to explore the apparent complex relation- 

ship of nutrition and cancer. The European Prospective 

Investigation into Cancer and Nutrition (EPIC), was 

designed to investigate the relationships between diet, nutritional 

status, lifestyle and environmental factors, and the 

incidence of cancer and other chronic diseases. 11 EPIC is 

the largest study of diet and health ever undertaken, having 

recruited over half a million people in ten European countries: 

Denmark, France, Germany, Greece, Italy, the 

Netherlands, Norway, Spain, Sweden, and the United 

Kingdom. Preliminary results of these large and well-conducted 

prospective EPIC studies published in 2005 indicated 

the absence of association between fruit and vegetable 

intake and incidence of breast cancer. 12 The authors emphasize 

that a small benefit can never be excluded and a modest 

benefit could exist for a subgroup of women (perhaps 

defined by genetic factors) or a subset of cases (for example, 

defined by estrogen receptor status). An important limitation 

is the lack of data on diet during childhood. If constituents 

of fruits and vegetables are acting to protect DNA 

from damage during childhood, nutritional intervention in 

utero and during early childhood may be a crucial factor in 

the phenotypic expression of cancer, and published studies 

could have entirely missed the critical periods. 

Although the findings on fruit and vegetable consumption 

and cancer may be disappointing, the epidemiological 

evidence clearly points towards lower risks of cardiovascular 

disease (CVD). The observation that dietary components 

such as saturated fatty acids are strongly correlated with the 

incidence of CVD in certain individuals but not in others 

points towards the genetic variations among individuals. 

CVD is a classic example of a diet-related chronic disease 

claiming more lives each year than the next 4 leading causes 

of death combined, which are cancer, chronic lower respiratory 

diseases, medical accidents, and diabetes mellitus. 13 

Preliminary mortality data show that CVD as the underlying 

cause of death accounted for 37.3% of all 2,440,000 

deaths in 2003 or 1 of every 2.7 deaths in the United States. 

CVD as an underlying or contributing cause of death 

(1,408,000 deaths in 2002) was about 58% of all deaths that 

year. Since 1900, CVD has been the no. 1 killer in the 

United States every year but 1918. Nearly 2,500 Americans 

die of CVD each day, an average of 1 death every 35 seconds. 

In addition, the aging of our population will result in 

an increased incidence of chronic CVD, including coronary 

artery disease (CAD), heart failure (CHF), and stroke. 14 

CVD is now understood to be an inflammatory disorder, 

and the relation between CVD and inflammation demonstrates 

the correlation between genes and nutrition factors. 

15 Current dietary recommendations for the prevention 

of CVD are derived from population-based epidemiological 

and observational studies and focus on the reduction of 

known cardiovascular risk factors such as low levels of 

HDL cholesterol and elevated levels of LDL cholesterol. 

The successful sequencing of the human genome and 

2 JANA Vol. 9, No. 1, 2006

understanding of the interactions between the human 

genome and the expressed phenotype provide us with new 

prevention and treatment strategies in the form of individualized 

dietary recommendation based on individualized 

genetic responses to dietary factors. As follows are several 

examples that validate this hypothesis. 

The enzyme methylene tetrahydrofolate reductase 

(MTHFR) is a crucial factor for the conversion of tetrahydrofolate 

into 5-methyl tetrahydrofolate, which is the active 

methylating form of folic acid. MTHFR is an integral component 

of the homocysteine cycle, providing the proper 

methyl group delivery through S-adenosylmethionine. 

Individuals with a homozygous form of the CT-677 polymorphism 

of MTHFR have, with normal dietary intake of 

folic acid, a decreased ability to produce 5-methyltetrahydrofolate 

from tetrahydrofolate, thereby decreasing the 

metabolism of methionine, an essential amino acid derived 

from dietary protein, and consequently increasing the level 

of homocysteine. In 1969, McCully made the clinical observation 

that linked elevated plasma homocysteine concentrations 

with vascular disease. 16 Hyperhomocysteinemia 

appears to be an independent risk factor for atherosclerosis 

and atherothrombosis. 17-18 Although severe hyperhomocysteinemia 

is rare, mild hyperhomocysteinemia occurs in 

approximately 5 to 7 percent of the general population. 19 

Patients with mild hyperhomocysteinemia have none of the 

clinical signs of severe hyperhomocysteinemia or homocystinuria 

and are typically asymptomatic until the third or 

fourth decade of life when premature coronary artery disease 

develops, as well as recurrent arterial and venous 

thrombosis. Several mechanisms have been suggested to 

explain the atherogenic capability of homocysteine: 

1. Homocysteine suppresses the expression of cellular glutathione 

peroxidase by endothelial cells, and this effect 

promotes lipid peroxidation by the reactive oxygen 

species formed during the oxidation of homocysteine, 20 

resulting in endothelial injury or dysfunction. 

2. Homocysteine is a potent mitogen for vascular smoothmuscle 

cells. 21 Exposure to homocysteine leads to a 

marked increase in vascular smooth-muscle proliferation 

in vitro, an effect that is due in part to an increase in the 

expression of messenger RNA of cyclin D1 and cyclin A. 22 

3. Homocysteine increases nitric oxide production in vascular 

smooth-muscle cells by activating the transcription 

factor NF-B. 23 Since NF-B/rel activity is essential for the 

proliferation of vascular smooth-muscle cells, 24 homocysteine-mediated 

activation of NF-B contributes to the 

mitogenic effect of homocysteine. 

Individuals affected by this CT-677 polymorphism may 

need augmented or higher levels of either 5-methyltetrahydrofolate 

or folic acid to limit their increased vascular risk 

factors and the potential progression to CVD. The American 

Heart Association has not yet recommended this personalized 

supplementation, 25 but the evidence clearly suggests 

that one integrate such a nutragenomic approach into the 

clinical practice. HDL cholesterol (HDL-C) levels play an 

important role in the modulation of cardiovascular disease 

risk profile. With the success of statins in lowering LDL 

cholesterol and reducing cardiovascular disease (CVD) 

event rate, interest is shifting towards HDL-C as a potential 

target for therapeutic intervention. High HDL-C is associated 

with low CVD event rate. Reverse cholesterol transport 

may contribute to this association, and describes transfer 

of cholesterol on HDL particles from peripheral tissues 

to the liver. The transport process involves: ATP-binding 

cassette transporter A1 mediated efflux of intracellular 

unesterified cholesterol to discoidal lipid-poor HDL particles; 

cholesterol esterification by lecithin; cholesterol acyltransferase; 

packaging of cholesterol ester into the particle 

core which becomes more spherical; and delivery of cholesterol 

ester to the liver for excretion in bile. The APOA1 

gene codes for the major protein in HDL-C and a particular 

variant of this gene in which adenosine (A) is substituted 

for guanosine (G) affects how the affected individual 

responds to polyunsaturated fatty acids (PUFAs). Clinical 

research revealed a significant sex difference. 26 HDL-cholesterol 

concentrations in women were associated with a 

significant interaction between polyunsaturated fatty acid 

(PUFA) intake as a continuous variable and APOA1 genotype 

(P = 0.005). When PUFA intake was 8%, HDL-cholesterol concentrations 

in carriers of the A allele were 13% higher than those 

of G/G subjects (P < 0.05). No significant allelic difference 

was observed for subjects in the range of PUFA intake of 

4–8% of energy. These interactions were not significant in 

men. This information can guide the clinician in selecting 

individualized PUFA supplementation in women at risk for 

CVD and to decide who will benefit from a diet rich in 

PUFAs and those who will not. The individualized PUFA 

supplementation is especially important, because some 

studies suggest that high intakes of linoleic acid may have 

adverse effects on proinflammatory cytokines and adhesion 

molecules. 27-28 Therefore the European Commission has 

recommended 4%-8% of energy from n-6 PUFAs, and the 

WHO recommends 5%-8% of energy from n-6 PUFAs. 

While the n-6 PUFA recommendations in North America 

and Europe are comparable, the Japan Society for Lipid 

Nutrition recommends that linoleic acid intake be reduced 

to 3%-4% of energy, and the International Society for the 

Study of Fatty Acids and Lipids (ISSFAL) also recommends 

a low level of linoleic acid intake (2% of energy and 

an upper limit of 3% of calories). 32 

Another important factor in the cholesterol metabolism 

relates to the interaction between animal fat and HDL-C. A 

specific gene (LIPC) determines the activity of the hepatic 

lipase enzyme, which then determines the binding and 

uptake of lipoproteins from the bloodstream. 

JANA Vol. 9 No. 1, 2006 3

A polymorphism in the hepatic lipase gene (LIPC), 

–514 (C/T) is associated with decreased hepatic lipase activity. 

33 Those individuals homozygous for this variant (TT 

genotype) have higher HDL-C levels than those with the 

normal gene expression (CC-genotype). 34-35 In examining 

the effects of the –514 CT LIPC polymorphism dietary fat 

interaction on HDL in 2,130 men and women participating 

in the Framingham Study, 36 the researchers found that the 

rarer TT genotype was associated with significantly higher 

HDL-cholesterol concentrations only in subjects consuming 

• Genetics: the study of the patterns of inheritance of specific 

traits, i.e. the study of single genes and their effects. 

• Genome: all the genetic material in the chromosomes of 

a particular organism; its size is generally given as the 

total number of base pairs. Explores which genes and proteins 

in the body are activated under different conditions 

and the influence of external factors (i.e. environment) on 

gene expression. 

• Genomics: the study of the functions and interactions of 

all the genes in the genome, including their interactions 

with environmental factors. 

• Genome projects: research and technology development 

efforts aimed at mapping and sequencing some or all of 

the genome of an organism. 

• Genotype: the genetic makeup of an organism, as characterized 

by its physical appearance or phenotype 

• Homozygous: having identical alleles for a single trait. 

• Metabolome: the sum of all metabolites. 

• Nucleotide: a unit of nucleic acid composed of phosphate, 

ribose or deoxyribose, and a purine or pyrimidine base. 

• Nutrigenetics: how genetic variations affect the interaction 

between nutrients and the health and disease potential of 

individual persons. 

• Nutrigenomics: the study of effect of nutrients on the 

genome, proteome, and metabolome. 

• Penetrance: probability of a gene or genetic trait being 

expressed. 

• Phenotype: the clinical presentation or expression of a 

specific gene or genes, environmental factors, or both. 

• Polymorphism: difference in DNA sequence among individuals. 

• Proteome: the sum total of proteins. 

• Single-nucleotide polymorphism (SNP): a common variant 

in the genome sequence; the human genome contains 

about 10 million SNPs. 

REFERENCES 

1. Lander ES, Linton LM, Birren B, et al. Initial sequencing and 

analysis of the human genome. Nature 2001;409:860-921. 

2. Graveley BR. Alternative splicing: increased diversity in the 

proteomic world. Trends Genet 2001;17:100-107. 

3. Wallace DC. Mitochondrial diseases in man and mouse. Science 

1999;283:1482-1488. 

4. Venter JC, Adams MD, Myers EW, et al. The sequence of the 

human genome. Science 2001;291:1304-1351. 

5. National Academy of Sciences. Diet, Nutrition and Cancer. 

National Academies Press: Washington, DC, 1982. 

6. World Cancer Research Fund and American Institute for Cancer 

Research. Food, Nutrition and the Prevention of Cancer: A 

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7. Davis CD, Milner J. Frontiers in nutrigenomics, proteomics, 

metabolomics and cancer prevention. Mutat Res 2004;Jul 

13;551(1–2):51-64 Review. Erratum in: Mutat Res 2005 Mar 

1;570(2):305 

8. Harris DM, Go VLW. How dietary components protect from 

cancer. Awad AB, Bradford PG, eds. Nutrition and Cancer 

Prevention. New York:Marcel Dekker, Inc:1991, 27-58. 

9. Omenn G S, Goodman G E., Thomquist MD, Balmes J Cullen, 

MR, Glass A, et al. Risk factors for lung cancer and for intervention 

effects in CARET, the Beta-Carotene and Retinol 

Efficacy Trial. J Natl Cancer Inst 88:1996;1550–1559. 

10. Albanes D, Heinonen OP, Hultunen JK, Taylor PR, Virtamo J, 

Edwards BK, et al. Effects of alpha-tocopherol and betacarotene 

supplements on cancer incidence in the Alpha- 

Tocopherol Beta-Carotene Cancer Prevention Study. Am J 

Clin Nutr 61:1995;1427S–1430S. 

11. European Prospective Investigation into Cancer and Nutrition, 

http://www.iarc.fr/epic/Sup-default.html. 

12. van Gils CH, Peeters PHM, Bueno-de-Mesquita HB, et al. 

Consumption of vegetables and fruits and risk of breast cancer. 

JAMA 2005;293:183-193. 

13. American Heart Association, Heart Disease and Stroke 

Statistics – 2006 Update. Circulation published online 

January 11, 2006. http://circ.ahajournals.org/cgi/reprint/CIR- 

CULATIONAHA.105.171600v1. 

14. Bonow RO, Smaha LA, Smith SC Jr, Mensah GA, Lenfant C. 

World Heart Day 2002: The international burden of cardiovascular 

disease: responding to the emerging global epidemic. 

Circulation 2002;106:1602-1605. 

15. Ross R. Mechanisms of disease: atherosclerosis — an inflammatory 

disease. N Engl J Med 1999 Jan;340:115-126. 

16. McCully KS. Vascular pathology of homocysteinemia: implications 

for the pathogenesis of arteriosclerosis. Am J Pathol 

1969;56:111-128. 

17. Kang SS, Wong PW, Malinow MR. Hyperhomocyst(e)inemia 

as a risk factor for occlusive vascular disease. Annu Rev Nutr 

1992;12:279-298. 

18. Stampfer MJ, Malinow MR, Willett WC, et al. A prospective 

study of plasma homocyst(e)ine and risk of myocardial infarction 

in US physicians. JAMA 1992;268:877-881. 

19. Ueland PM, Refsum H. Plasma homocysteine, a risk factor for 

vascular disease: plasma levels in health, disease, and drug 

therapy. J Lab Clin Med 1989;114:473-501. 

20. Upchurch GR Jr, Welch GN, Fabian AJ, et al. Homocyst(e)ine 

decreases bioavailable nitric oxide by a mechanism involving 

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JANA Vol. 9, No. 1, 2006


The Physiological and Psychological 

Effects of Animal Therapy: 

Examining the Healing Power of Animals 


Niki Sepsas,* Associate Editor, Allen Montgomery, RPh, Managing Editor 

Journal of the American Nutraceutical Association 

Birmingham, Alabama 

Growing up with an affectionate, furry friend is a very 

satisfying childhood memory for many, as the love and care 

we bestowed on our pets was returned to us via their unconditional 

love and devotion. In recent years, however, a 

growing number of people have begun touting more than 

just the “warm and fuzzy” benefits of interacting with the 

animals in our lives. Medical practitioners, veterinarians, 

health care professionals, and others are pointing to the 

potential therapeutic value of this ancient bond between 

people and animals. 

In November 2005, the American Heart Association 

released the results of a clinical study that confirmed what 

many have suspected for years—interaction with pet therapy 

teams has a measurable positive effect on hospital 

patients. The results of the study, Animal Assisted Therapy 

and Heart Failure, 1 were presented at the American Heart 

Association’s “Scientific Sessions in Dallas” gathering. 

Researchers discovered that a 12-minute visit with “man’s 

best friend” helped heart and lung function by lowering 

pressure, diminishing release of harmful hormones, and 

decreasing anxiety among hospitalized heart failure 

patients. Benefits exceeded those that resulted from a visit 

with a human volunteer or from being left alone. 

The therapeutic approach of using dogs to soothe people’s 

minds and improve health has been considered more a 

“nicety” than credible science, according to Kathie M. 

* Corresondence: 

Niki Sepsas 

1057 Sherbrooke Drive 

Birmingham, AL 35205 

Phone: (205) 942-5335 

Email: nikisepsas@bellsouth.net 

Cole, RN, MN, CCRN, a clinical nurse at the UCLA 

Medical Center in Los Angeles and lead author of the study. 

To provide clinical proof of the benefits of animal-assisted 

activities and therapy, researchers studied the reactions of 

76 hospitalized heart failure patients to a visit from either a 

human and dog volunteer team, a human volunteer alone, or 

no visit (the at-rest group). 

During the 12-minute intervention with the volunteer/dog 

team group, specially trained dogs would lie on 

patients’ beds so they could touch them while interacting 

with the volunteer/dog team. Researchers monitored 

patients’ homodynamics – the collective system of measurement 

for blood volume, heart function, and resistance 

of the blood vessels. Investigators also measured epinephrine 

and norepinephrine levels at three time periods, before, 

during, and after the interaction, and administered an anxiety 

test before and after as well. 

Researchers found that anxiety scores dropped 24 percent 

for participants who received a visit from the volunteer/dog 

team. Scores for the volunteer-only group dropped 

10 percent while the at-rest group’s score did not change. 

Levels of the stress hormone epinephrine decreased an 

average of 14.1 picograms/wL, or 17 percent, in the volunteer/dog 

team group; 2 percent in the volunteer-only group; 

and rose an average of 7 percent in the at-rest group. 

The pulmonary capillary wedge, a measurement of left 

atrial pressure, dropped an average of 10 percent at the end 

of the interaction for those receiving volunteer/dog team 

therapy. It increased 3 percent for the volunteer-only group, 

and increased 5 percent for the at-rest group. 

Systolic pulmonary pressure, a measure of pressure in 

the lungs, dropped 5 percent during and 5 percent after 

therapy in the volunteer/dog team group. It rose during and 

after therapy in the other two groups. 

7

The initiative was one of the first studies to use scientific 

measurements to demonstrate that therapeutic dogs 

lower anxiety, stress, and heart and lung pressure among 

heart failure patients. “The study demonstrates that even a 

short-term exposure to dogs has a beneficial physiological 

and psychological effect on patients who want it,” Cole said. 

“This therapy warrants serious consideration as an adjunct 

to medical therapy in hospitalized heart failure patients. 

Dogs are a great comfort. They make people happier, 

calmer, and feel more loved. This is huge when patients are 

scared and not feeling well.” 

Other studies and findings add to the mounting volume 

of scientific research that points to the benefits of animal 

therapy: 

• In 2002, the Journal of the American Medical Association 

reported that children exposed to pets during the first year of 

life have a lower frequency of allergic rhinitis and asthma. 

• In her article “Children’s Adjustment to the Death of a 

Parent,” published in the Journal of Youth and 

Adolescence, Victoria H. Raveis noted that the companionship 

of pets (particularly dogs) helps children adjust 

better to the serious illness or death of a parent. 

• In 2002, a study of autistic children by Rehabilitation 

Services of Roanoke, Virginia, revealed that animal-assisted 

therapy is more effective than conventional therapy. 

These findings and the results of the UCLA study are 

in line with what many healthcare professionals across the 

country are observing in their own practices. 

Patti Wilson works in a world where fear, sorrow, and 

pain are everyday occurrences. A nurse practitioner in the 

Department of Radiology of the Cancer Center at the 

University of Alabama at Birmingham (UAB) Hospital, she 

reports for duty each day in an environment beyond comprehension 

for most of us. She regularly witnesses the emotional 

trauma of patients and families battling cancer, the 

number two cause of death in the United States. 

In addition, Wilson’s work puts her in a position to 

observe the remarkable therapeutic benefits these people 

receive from specially trained animals and their handlers. 

“I have seen eyes dulled by fear and pain become bright 

with joy when these sweet animals sit patiently and look 

back with bright, loving eyes, and when the human component 

of the team takes the time to show they care,” Wilson 

states. “I’ve seen the hands of patients, family members and 

staff reach out to pet these precious animals and have seen 

the smiles, . . . smiles that replace the frowns of those who 

are very ill and anxious.” 

But smiles are not the only benefits, as Wilson notes 

and the UCLA study points out. “Physiologically, the benefits 

include a decrease in hypertension and in cortisol 

steroids, the hormones responsible for the harmful effects 

of stress,” Wilson continues. “Psychologically, it has been 

8 

HIP volunteers and their therapy dogs working 

with young patients undergoing treatment at UAB’s 

Comprehensive Cancer Center in Birmingham. 

documented to reduce feelings of depression and increase 

feelings of well being and confidence.” 

Wilson’s findings are echoed by others. 

“You could literally watch her blood pressure lowering 

on all the monitors while Drumm, the therapy dog, was 

there visiting,” reports Jenni Boswell, a pet therapy volunteer 

after a visit to the Cardiac Unit of University Hospital 

in Birmingham, Alabama. 

A desire to bring this documented and remarkable 

healing power of pets to those in need led one Birmingham 

woman to establish a local organization that has since been 

spotlighted on national and international stages. 

Born into an Air Force family, Alabama native Beth 

Franklin called many places home while she was growing 

up. Her professional life with Blue Cross Blue Shield of 

Alabama spanned 21 years before she decided to put her 

Master’s degree in counseling to work in other areas. A 

lifelong love of pets led her to a volunteer position with the 

Birmingham Humane Society, where she eventually 

became its director. While participating in a program that 


took certain of the shelter’s animals to nursing homes, 

Franklin became interested in expanding that initiative into 

a full-blown pet therapy program. 

“Taking our animals to these homes was undeniably beneficial 

to the residents, but it was unpredictable since the animals 

we were working with had been living in a shelter,” 

Franklin explains. “I decided that a formal program was needed 

to train the pets and the people handling them to prepare 

them for working with hospitalized patients and the elderly.” 

Franklin’s research led her to the work of the Delta 

Society, ® an international not-for-profit organization that 

aims to improve human health through service and therapy 

animals. The Delta Society ® was established in 1977 in 

Portland, Oregon, under the leadership of Michael 

McCulloch, MD. The society’s first president, Leo Bustad, 

DVM, PhD, was the dean of a veterinary college and pioneer 

in human-animal bond therapy and application. His 

focus and that of the society’s early founders was on funding 

the first credible research on why animals are so important 

to the general population and specifically how they 

affect health and well-being. 

More than 20 studies funded by the Delta Society ®2 

have now established that connection. The organization 

presently concentrates its efforts on creating educational 

materials to apply the resulting scientific information in 

everyday life and to provide direct services at the local level. 

Their Standards of Practice on Animal Assisted Activities 

and Animal Assisted Therapy provides guidance in administrative 

structure of Animal Assisted Activities and Therapy 

(AAA/T) programs, including animal selection, personnel 

training, treatment plan development, and documentation. 

Using the Delta Society ® guidelines, Franklin founded 

Hand In Paw (HIP) in 1996 in Birmingham, Alabama. The 

organization achieved 501-c-3 status in 1997, and Franklin 

serves as the executive director. 

Franklin wasted little time putting her goals into action. 

She understood that well-trained volunteers and animals are 


HIP founder, Beth Franklin 

fundamental to establishing a series of programs that were to 

focus first on hospitalized children. She worked diligently 

to design and implement an HIP Pet Partner ® initiative that 

trained an animal and its handler as a team that would visit 

children in Birmingham’s medical treatment facilities. 

Jessica, her own toy poodle, and Maggie, a Shih-tzu, were 

the first two Delta Society registered animals in Alabama. 

“I targeted UAB’s Comprehensive Cancer Center,” she 

notes. 3 “I felt we could really make a difference with children 

who are undergoing cancer treatment. We were readily 

accepted as Louis Josof, the Cancer Center’s director, is 

an advocate for our program. It took a while for us to 

receive clearance from Children’s Hospital as there are liability 

concerns, infection control procedures, and other 

issues. We began our visits at the UAB Comprehensive 

Cancer Center in 1996, and at Children’s Hospital the following 

year. The Pet Partner ® program took off right away. 

Children in a cancer ward are understandably frightened 

about their surroundings, the treatment they are undergoing, 

and the people around them. Maggie and Jessica were 

perfect in their roles of reaching those children.” 

Nurse practitioner Patti Wilson surveyed members at 

the Cancer Center as to the effectiveness of HIP’s work. 

“One of the nurses said she had seen children who would 

normally need anesthetic to undergo radiation treatment not 

need the anesthesia after a visit from a HIP team,” Wilson 

states. “This is amazing and something in which HIP can 

take great pride.” 

HIP Pet Partner ® teams were found to provide an 

invaluable calming influence on children tearful because of 

the physical and emotional trauma of receiving radiation 

therapy. Animals were even placed next to the youngsters 

on the table as they were sedated. More than a few healthcare 

professionals have admitted to shedding tears as a 

child fitted with a tight face mask in preparation for sedation 

held on to her mother with one hand and to a therapy 

animal with the other. 

The success of Franklin’s program at UAB’s 

Comprehensive Cancer Center soon led to calls from other 

medical facilities. Pet Partner ® teams began making regular 

visits to the Department of Outpatient Rehabilitation 

Services at Birmingham’s Children’s Hospital. 

Franklin quickly ratcheted up her efforts to train additional 

animals and volunteers. The dedicated individuals 

and trained therapy animals underwent testing and registration 

through the Delta Society, and were re-tested regularly 

according to HIP advanced criteria. 

Hand In Paw Pet Partner ® teams now regularly visit 

UAB, Children’s, Brookwood, St.Vincent’s, and Baptist 

Medical Center - Montclair Hospitals in the greater 

Birmingham area, which has become nationally and internationally 

recognized as a premier center for medical treatment 

and research. HIP’s success at these healthcare facil- 

9

Glory, a two-year old Sheltie, in Birmingham, Alabama, 

works with toddler at the Bell Center for Early Intervention 

Programs, which focus on developmentally delayed children. 

ities led Franklin to launch other innovative programs targeting 

additional groups. 

HIP began sending Pet Partner ® teams to the locked 

psychiatric unit at Children’s Hospital to visit patients age 

12 to 18 who had been admitted for suicide attempts, psychosis, 

aggressive behavior, mood instability, sexually “acting 

out” behavior, eating disorders, depression, and alcohol 

and drug abuse. 

“I’ve seen an individual smile for the first time since his 

admission just at the sight of (animal therapy dogs) Molly, 

Sarge and Calypso,” states Lisa Trentham, an occupational 

therapist previously at the unit. “I’ve also witnessed kids 

talking expressively to one of the pets when I’d been unable 

to facilitate such interaction, and I’ve seen kids who have 

been too depressed to get out of bed make an effort to attend 

group session just to see the pets.” 

Trentham also feels that therapy pets are extremely 

effective in working with children who have abused animals. 

These children may have sacrificed animals for supposed 

‘religious’ beliefs, are involved with dog fighting, 

have killed the family pet due to command hallucinations or 

to get parental attention, have put their pet in the microwave 

or dryer, or have branded an animal with a gang sign. 

“To watch individuals with a history of animal abuse 

pet these animals with gentle hands and look into their eyes 

Therapy dog Molly working with toddler undergoing 

treatment at Children’s Hospital in Birmingham, 

Alabama. 

and see reflected back a bit of compassion is a wonderful 

sight to behold,” she adds. “Considering the seriousness of 

these actions, I view myself and everyone involved with 

animal-assisted therapy as steps in the ladder to helping 

overcome animal cruelty. And the pets don’t just reach the 

children, they reach the staff as well. Assisting with the 

burnout that can occur among mental health professionals 

is an added benefit of animal-assisted therapy.” 

HIP is also involved in an experimental neuromotor 

rehabilitation program that has attracted international attention 

due to its success. UAB’s Pediatric Neuromotor 

Research Clinic launched ACQUIREc, an exciting new therapy 

to assist children with neuromotor disorders resulting 

from cerebral palsy, traumatic brain injury, and other neurological 

injuries. The acronym embodies the program’s concept 

of Acquisition of new motor skills through Continuous 

practice and shaping to produce Quality movement of the 

Upper extremity through Intense therapy and Reinforcement 

in Everyday patterns and places. The subscript “c” indicates 

the important component of casting (as a way of immobilizing 

a body part so that its complement can be strengthened). 

Therapy involves three major components: 

• Therapy must be given many hours a day for many consecutive 

days to produce changes in the brain and motor 

behavior. 

• The weaker upper extremity must be facilitated and 

trained in many specific tasks appropriate to the child’s 

behavior. 

• The stronger upper extremity must be completely 

restrained for an extended time. 

The program’s administrators felt that HIP could play an 

important role in inducing the children to perform tasks with 

their weaker extremity in the presence of therapy animals. 

Stacie Williams, an occupational therapist previously 

with the ACQUIREc program, was working with four-year 

old Tyler, whose parents had come to UAB from their home 

in Pennsylvania to work on improving the fine-motor skills 

of Tyler’s right hand. “His less-affected left arm and hand 


were in a cast,” Williams explains, “and we discussed how 

the therapy pet might help Tyler focus on using his right 

hand, particularly his fingers. During our session, we took 

a walk and Tyler used his right hand to hold the leash as I 

guided them. When we returned, I asked him to remove the 

dog’s leash. I showed him how to use his finger and thumb 

to release the leash, and we all clapped when, after several 

attempts, he did it himself. He also practiced taking brightly 

colored pom-poms out of a box to ‘decorate’ the dog.” 

Williams also worked with six-year-old Emily, who had 

suffered a stroke in utero. Before ACQUIREc, Emily was 

exclusively using her right hand. At first, she would turn her 

left palm up only with assistance, but in order to give the 

therapy pet a treat, she soon did it on her own. She also decorated 

the dog’s fluffy hair with pom-poms, picked them 

up, and returned them to their box. 

Williams reports that both Tyler and Emily have now 

completed their 21-day treatment and accomplished their 

goals. They join children from across the country and 

around the world who have found this new approach to 

neuro-rehabilitation extremely beneficial. 

In addition to the Petscription program that provides 

services at medical, psychiatric, and rehab hospitals, social 

service agencies, and at camps for children with serious 

chronic illnesses, a number of other HIP initiatives are bringing 

hope and light where before there had been darkness: 

The Pawsitive Living program teaches positive living 

skills and anger management to high-risk youth by using 

therapy animals to help teach respect and empathy. Often 

this program intervenes with youth who have displayed violence 

toward animals in the past, an early sign of potential 

violence towards people. The Pawsitive Living program 

serves agencies such as Glenwood Mental Health, Salvation 

Army Youth Services, Family Court of Jefferson County, 

and the YWCA Family Violence Center. 

“In my years as Presiding Circuit Judge of Jefferson 

County Family Court, I have seen many programs aimed at 

helping at-risk youth,” states Sandra H. Storm. “Hand In 

Paw’s Pawsitive Living is one that had a beneficial impact 

on at-risk teens from Jefferson County Family Court. The 

difference in our clients’ attitudes upon completion of the 

program was remarkable.” 

Pawsitive Living programs have had a similar impact 

on boys at Birmingham’s Daniel House, a facility run by 

Glenwood Mental Health. The 12-week sessions teach the 

youngsters how to set goals for themselves through challenges 

and problem solving involving therapy pets. The 

children discuss subjects such as self-esteem, anger management, 

abandonment, respect for others, and diversity. 

The pets and their handlers provide an opportunity for 

children to open up and express thoughts and feelings (that 

many have never before discussed) with someone who will 

listen without criticizing. 


“The Pawsitive Living kids inspire me to hope, to 

dream, to believe in a better world,” states Sheila Cavallo, 

the program’s facilitator and manager. “Their faith and 

hope are the foundation of my work. Their willingness to 

trust, even in the wake of years of abuse, is a tremendous 

inspiration to me, proving to me again and again that the 

words ‘lost cause’ never belong in the same sentence with 

the word ‘child’. 

HIP’s Sit, Stay, Read program pairs reluctant readers 

from five to twelve years old with therapy animals to make 

reading fun, non-threatening, and a positive learning experience. 

Each child reads to his animal one-on-one for 20 to 

30 minutes. Part of the magic of this program is the quiet, 

calming effect of a peaceful animal. Dogs can be petted 

and loved before, during, and after the reading session. 

They ease conversation and relax the children as volunteers 

have a chance to ask about the child’s day, share a laugh, 

and be a warm, caring friend. 

“Overall, the most dramatic change has been the 

increased excitement and enthusiasm toward reading,” 

states Rhonda L. McKay, After School and Summer 

Enrichment Coordinator for the YWCA. 

Pet Partner ® teams work at the Bell Center for Early 

Intervention Programs, which focus on developmentally 

delayed children from eighteen months to three years old. 

HIP handler and therapy dog teams have participated in 

rape survivors’ retreats where they make a significant 

impact on women dealing with the trauma of sexual assault. 

HIP teams helped cheer families displaced by 

Hurricane Katrina who were applying for aid at 

Birmingham’s Disaster Recovery Center. Many evacuees 

had lost family members, homes, and beloved pets. Visits 

from Pet Partner® teams helped children cope with drastic 

change and to adapt to their new surroundings. 

A Paws for Comfort program was launched in 2005 to 

provide to military families respite from the anxiety and 

worry that accompany long separations from loved ones 

serving overseas. “Paws for Comfort was developed to provide 

healing comfort and an effective vehicle for expression 

of grief after crises,” says Beth Franklin. 

Today, Hand in Paw has more than 80 Pet Partner ® 

teams bringing the healthful benefits of animal therapy to 

thousands of people in 35-plus facilities in the four counties 

surrounding Birmingham. Beth Franklin’s staff has grown 

to seven full-time employees supported by a huge network 

of volunteers and their therapy pets. In addition to dogs, 

HIP therapy animals include cats, a pony, a miniature horse, 

a bird, a rabbit, and a rat. 

The organization receives no state or federal funding, 

but relies instead for their operating revenue on fund-raising 

activities and donations and grants from numerous 

foundations. A recent grant from actor Paul Newman helps 

11

underwrite the Pawsitive Living program as a sentencing 

option for Jefferson County Family Court. Newman’s support 

of the program is a powerful endorsement of HIP by a 

leading national philanthropist. 

Other local organizations and foundations that support 

the work of HIP include the Children’s Trust Fund of 

Alabama, Hill Crest Foundation, the Community 

Foundation of Greater Birmingham, and the Junior League 

of Birmingham. 

Franklin and her staff have also launched a number of 

creative and innovative fund-raising initiatives to generate 

local support for HIP. The organization’s Picasso Pets Debut 

and Auction, supported by a long list of local businesses and 

individuals, raises thousands of dollars each year through the 

auctioning of artwork “painted” by animals owned by prominent 

members of the community. The event also showcases 

to the public the many therapy pets and their handlers. 

The work of the dedicated staff of HIP in the 

Birmingham area has served as a model for others seeking 

to develop similar protocols and programs around the country. 

In November 2004, a group of occupational therapists, 

physical therapists, and others who are involved in animal 

assisted therapy made a trip to Birmingham from El Paso, 

Texas, to learn how HIP incorporates AAT in early intervention 

therapy for developmentally delayed children. In 

their national search, HIP was the only organization they 

could find in the country that combined AAT with early 

intervention therapy. 

That same month, Debbie Coultis, president of PAN 

(People, Animals, and Nature), visited and observed HIP 

programs to assess the Pawsitive Living curriculum. Her 

assistance in getting the PL curriculum into a format that 

can be distributed nationally will enable HIP to share this 

unique program for reaching troubled youth on a national 

and, possibly, a worldwide basis. 

HIP staff and volunteers recently welcomed visitors 

from Japan interested in expanding animal-assisted therapy 

programs to address behavioral and medical problems in 

their country. 

“Our primary focus now is to expand the services we 

offer the community through the development of new relationships 

with schools, demographic groups and others in 

need,” states Rachel Reinhart, HIP’s Program Manager. 

“We are blessed to have a wonderful staff of volunteers 

eager to help us reach more people. Hand In Paw has been 

extremely successful in developing programs that are having 

a very real and dramatic impact on the lives of thousands 

of people in our area and beyond.” 

The work of Beth Franklin and her dedicated staff has 

been recognized both locally and nationally. Recently, 

Franklin and her therapy dogs were honored with the 2005 

Delta Society Beyond Limits ® Award for Lifetime 

Achievement. She was recognized both for her work as a 

Pet Partner ® and for founding and leading Hand In Paw. 

In addition, HIP Pet Partner ® teams received two of 

only five 2004 Beyond Limits Awards presented nationwide 

by the Delta Society honoring the very best among those 

working in fields related to the human-animal bond. 

Beth Franklin’s greatest rewards come in seeing the 

difference that she, her staff, the organization’s many volunteers 

and their pets are making in the lives of so many 

people. She recounts a particularly moving experience: 

“At a visit to Children’s Hospital, one of our deaf therapy 

dogs and his handler were visiting a deaf child,” she 

relates. “The child, previously withdrawn and downtrodden, 

was elated at meeting a friend who could ‘speak’ her 

language. Hearing the child squeal and seeing her jump up 

from her wheelchair, painted a memory I will never forget.” 

For additional information on Hand In Paw, 

including volunteer opportunities and 

donations, contact: 

Beth Franklin 

Hand In Paw 

2616 7th Avenue South 

Birmingham, AL 35233 

(205) 322-5144 

info@handinpaw.org 

www.handinpaw.org 

REFERENCES: 

1. American Heart Association Annual Meeting, November 

11, 2005, Dallas, Texas. http://www.americanheart.org/presenter.jhtml?identifier=3035327. 

2. Delta Society. 875 124th Ave NE, Ste 101, Bellevue, WA 

98005. www.deltasociety.org. 

3. Westlake-Kenny, B. Pet Therapy: A Healing Hand in Paw. 

UAB Magazine 1999 winter; 19:1:1. Accessible online at 

http://main.uab.edu/show.asp?durki=31686. 


R E V I E W A R T I C L E 

Microdose DNA for the Treatment of Acute and 

Chronic Respiratory Diseases and Otitis Media 

Acute and chronic respiratory diseases such as common 

colds, allergic rhinitis, sinusitis, bronchitis, mucositis, 

asthma, emphysema, chronic obstructive pulmonary disease 

(COPD) and cystic fibrosis, as well as otitis media, 

have different etiologies. Causative agents include microorganisms 

such as viruses (colds) and bacteria (otitis), environmental 

insults such as cigarette smoke (COPD), radiation 

therapy (mucositis) and gene mutations (cystic fibrosis). 

However, these diseases share some common clinical 

features, including inflammation, bronchial and/or sinus 

congestion, obstructed airflow, and the production of large 

amounts of sputum and/or excessive nasal mucus. Many of 

these diseases may also have common physiological and 

immunological characteristics. 1-19 

Cystic fibrosis (CF) is an often-fatal genetic disorder of 

exocrine function characterized by abnormally viscous mucus 

secretions. Such secretions precede chronic pulmonary 

obstruction, pancreatic insufficiency and elevated sweat sodium 

and chloride levels. The viscosity of sputum produced by 

CF patients is believed to result from a high content (approximately 

10% of the total sputum dry weight) of DNA released 

from necrotic neutrophils in the sputum. 2,12,14,16,20-22 This 

observation has led to the use of DNase (Dornase alfa; 

Pulmozyme) as a CF therapy in conjunction with the antibiotics, 

bronchodilators and corticosteroids regularly used in 

the treatment of CF. 23-25 The rationale for such therapy is 

that degrading DNA in sputum reduces the viscosity of the 


John McMichael, PhD 

The Institute for Therapeutic Discovery 

P.O. Box 127 

Delanson, NY 12053 

Phone: 518-872-1144 Fax: 518-872-0753 

Stephen W. Mamber, PhD 

John McMichael, PhD* 

The Institute for Therapeutic Discovery 

Delanson, New York 

sputum and results in an increased ability of the patient to 

evacuate sputum from the lungs and nasal passages. 23-25 

As the presence of neutrophil DNA in the sputum of 

CF patients suggested an aberrant compensatory immune 

response, it was hypothesized that mammalian DNA itself 

could be employed as a neutralization therapy. This hypothesis 

became the basis for the development of a sublingually 

administered therapeutic composed of a proprietary formulation 

of DNA fragments derived, initially, from calf thymus 

DNA. 26 The hope was that, in the case of CF, sublingual 

dosing with DNA would ultimately lead to decreased 

neutrophil necrosis and DNA release into the sputum in the 

lungs, which would in turn result in decreased sputum viscosity. 

26 In evidence-based clinical testing, the DNA therapeutic 

was successful at reducing mucus viscosity and/or 

decreasing mucus accumulation in the respiratory tracts of 

a number of CF patients. Subsequently, this sublingual 

therapeutic approach was extended to respiratory diseases 

other than CF. While the specific mechanism(s) of action 

are still being elucidated, it was found that the DNA therapeutic 

had broader application to a variety of acute and 

chronic respiratory diseases as well as to the treatment of 

otitis media. 27-31 Both calf thymus DNA and salmon sperm 

DNA have been used in clinical testing. The salmon sperm 

DNA was found to have therapeutic activity equivalent to 

that of the bovine derived DNA. In addition to eukaryotic 

vertebrate DNA, prokaryotic (bacterial) DNA and synthetic 

DNA have also been evaluated for activity in treating various 

respiratory ailments. It is speculated that the source 

of DNA may be less consequential than the method by 

which microdose DNA is formulated. 

Milkhaus Laboratory, Inc., developed the original product, 

named HP-3 or ML-03, using eukaryotic vertebrate 

DNA. HP-3/ML-03 was active in three separate FDAapproved, 

placebo-controlled, double-blind Phase II clinical 

JANA Vol. 9, No. 1, 2006 13

trials for the treatment of chronic bronchitis, COPD and CF. 

The current DNA-based therapeutic, derived from salmon 

testicle DNA, is being sold commercially as Mucolyxir. TM It 

has been referred to as microdose DNA because it is administered 

in microgram-range doses. The purpose of this article 

is to describe the product, discuss possible mechanisms 

of action and summarize clinical trial results. Evidencebased 

clinical experiences with microdose DNA as a treatment 

for a variety of respiratory conditions as well as otitis 

media have been described in detail in the patent literature 

and are summarized in a separate article. 26-32 

PRODUCT DESCRIPTION 

DNA for use in MucolyxirTM is eukaryotic DNA, 

specifically salmon testicular DNA obtained from a commercial 

source. The DNA is solubilized in sterile phenolated 

saline, and proprietary methods are used to generate a 

mixture of DNA fragments within a definable range of molecular 

weights. It is administered as sublingual drops. 

Although previous clinical investigations with microdose 

DNA employed solutions of 12 ug/ml (about 0.6 ug/dose 

based on a drop volume of about 50 ul), MucolyxirTM is formulated 

in its liquid vehicle at a concentration of 6 ug/ml 

(approximately 0.3 ug/dose). A typical administration regimen 

is 1-2 sublingual drops one to four times daily. 

PROPOSED MECHANISMS OF ACTION 

Reduced congestion, decreased mucus viscosity and/or 

decreased inflammation in the upper and lower airways leading 

to improved respiratory functions are common observations 

in patients treated with microdose DNA. Based on 

dosage (low-level) and administration route (generally sublingual), 

microdose DNA may promote or restore homeostasis 

within the respiratory tract by an as yet uncharacterized 

signaling pathway(s) involving the body’s regulatory systems, 

notably (though not limited to) the immune system. 

There have been several proposed hypotheses regarding specific 

mechanisms by which microdose DNA can alleviate 

various respiratory ailments. While each of these hypotheses 

is considered separately, one cannot exclude the possibility 

of multiple and/or interrelated and/or other mechanisms 

being responsible for the observed effects. 

1. Immunotherapy via Desensitization, 

Hyposensitization or Neutralization 

The original concept, proposed in the early 1990s, for 

utilizing a low dose of exogenous mammalian DNA as a 

treatment for CF was that it could generate an uncharacterized 

molecular signal that would trigger a localized 

immune response to alleviate the buildup of DNA in the 

sputum of afflicted individuals. The rationale for this 

approach was based on principles of allergy immunotherapy, 

in which repeated administration of a potential allergen 

would result in a decreased immune or inflammatory 

response. 33-38 The net effect would be reduced disease 

symptoms, i.e., reduced sputum/mucus production, inflammation 

and airway obstruction. It was hypothesized that 

the DNA released from necrotic neutrophils was itself acting 

as an allergen of sorts, promoting inflammation in CF 

patients. One problem with this hypothesis was that microdose 

DNA was found to be effective in treating respiratory 

diseases other than CF. Interestingly, however, the potential 

of microdose DNA as an immunotherapeutic substance 

may actually have been ahead of its time, as the concept of 

using specific immunostimulatory DNA sequences (CpG 

DNA) for the immunotherapy of acute and chronic respiratory 

diseases did not appear in the literature until the latter 

part of the decade and into the 21 st century. 39-41 

2. Stimulation of Chloride Secretion and Mucociliary 

Clearance via Interaction with P2 Receptors 

Although DNA in the sputum of CF patients increases 

its viscosity, exogenous purine and pyrimidine nucleosides 

such as ATP and UTP can, by interacting with P2 receptors 

and stimulating chloride secretion, improve mucociliary 

clearance in CF and other respiratory ailments characterized 

by excessive sputum production. 42-45 Subsequently, it 

was hypothesized that microdose DNA may alleviate respiratory 

conditions such as CF and bronchitis by interacting 

with P2Y receptors responsible for stimulating mucociliary 

clearance within the respiratory tract. P2Y receptors represent 

a family of 7-transmembrane G-protein–coupled 

receptors. Purine and pyrimidine nucleotides/nucleosides 

are known ligands for this family of receptors. 46-48 A specific 

P2Y receptor, P2Y(2), is present in a variety of airway 

epithelial cell types, such as the ciliated epithelial cells and 

goblet cells of the trachea and bronchi, as well as in middle 

ear ciliated epithelial cells. 49-51 Activation of P2Y(2) 

appears to result in increased mucociliary clearance in the 

lungs and other respiratory tract tissues and in the middle 

ear. 51-53 P2Y(2) receptor agonists, notably nucleoside 

triphosphates such as adenosine 5’-triphosphate (ATP) and 

uridine 5'-triphosphate (UTP) and related analogs, have 

been shown to promote a variety of activities related to 

improved mucociliary clearance. This includes stimulation 

of chloride secretion in human airway epithelial cell cultures, 

stimulation of mucin secretion in human nasal and tracheobronchial 

tissue explants, modulation of ion transport 

in an in vitro middle-ear epithelial cell line, increased cilia 

beat frequency in human airway epithelial cells in vitro, and 

increased tracheal mucus velocity, whole-lung mucociliary 

clearance and cough clearance in animals and in human respiratory 

disease patients. 42-43,45,51-60 However, when tested in 

vitro, it was determined that microdose DNA was not acting 

as a P2Y(2) receptor agonist (unpublished data). The possibility 

that microdose DNA could be affecting P2 receptors 

other than P2Y(2) requires further investigation. 


3. Generation of Beneficial Immune Responses through 

Changes in Th1/Th2 Cytokine Ratios 

Another viable hypothesis for the beneficial effects of 

microdose DNA in treating various respiratory diseases is 

that it is acting as an immunostimulant capable of altering 

certain cytokine imbalances. This hypothesis reflects on the 

original rationale for microdose DNA usage–the generation 

of a molecular signaling pathway that would result in a 

favorable immune response. 

The immunostimulatory effects of bacterial extracts, 

specifically extracts of Mycobacterium bovis bacillus 

Calmette Guerin (BCG), and their application to host 

defense have been investigated since the late 19th century. 

Eventually (about a century later), it was determined that 

unmethylated nucleotides derived from the BCG DNA that 

contained a specific nucleotide sequence, CpG, were 

responsible for the observed beneficial immunostimulatory 

effects. 39-40, 61-63 Oligodeoxynucleotides (natural or synthetic) 

containing the CpG motif, referred to in the literature as 

immunostimulatory DNA sequences, can be readily detected 

by vertebrate innate immune receptors (toll-like receptors), 

including those on dendritic cells, macrophages, 

monocytes and neutrophils. 62,64-68 Toll-like receptor signaling 

by immunostimulatory DNA sequences leads to 

changes in cytokine production. 69-74 In respiratory ailments 

such as viral infections and allergen-induced respiratory 

inflammation, the ratio of type-1 and type-2 T-helper cells 

(Th1/Th2 ratio) is altered. Specifically, the Th2-type 

humoral immune response is enhanced, while the Th1-type 

cell-mediated immune response may be suppressed, leading 

to unfavorable alterations in cytokine balances that favor a 

given respiratory disease state. 75-80 Immunostimulatory 

DNA has been shown to balance and enhance the immune 

response via normalization of Th1/Th2 cytokine ratios. 

Consequently, the Th2 response is reduced and/or the Th1 

response is enhanced to restore homeostasis in ailments 

such as respiratory syncytial virus infections, asthma and 

allergies that affect the respiratory tract. 61-62, 81-88 In vivo, 

immunostimulatory DNA administered mucosally or systemically 

has been shown to reduce inflammation and inhibit 

airway remodeling and airway hyper-responsiveness in a 

rodent allergic rhinitis model. 89 In a primate model of allergic 

asthma, airways from nucleotide-treated monkeys had 

thinner basement membranes, fewer mucus cells, fewer 

eosinophils and fewer mast cells than controls. 90 

A key issue with the hypothesis that microdose DNA 

has immunostimulatory effects analogous to that of CpG 

DNA is that the former is derived from eukaryotic DNA 

rather than from prokaryotic DNA. Not only does eukaryotic 

DNA contain significantly fewer CpG motifs than does 

prokaryotic DNA, there is more DNA methylation. The 

presence of relatively few unmethylated CpG motifs in 

eukaryotic DNA makes it less immunostimulatory than 

prokaryotic DNA. 63, 84, 91-95 However, it is speculated that 


the resulting combination of DNA sizes and sequences 

resulting from product preparation may be capable of generating 

a beneficial immune response to alleviate the abovenamed 

respiratory diseases in a manner analogous to that of 

Immunostimulatory DNA of prokaryotic origin. Thus, 

more in vitro and in vivo research with microdose DNA is 

required in order to validate the hypothesis that 

MucolyxirTM can act as an immunostimulatory agent in the 

manner of CpG DNA. 

4. Antiinflammatory Effects Such as Decreased 

Production of Proinflammatory Cytokines Increased 

Production of Antiinflammatory Cytokines 

A fourth hypothesis is that microdose DNA can ameliorate 

various respiratory diseases by acting as an antiinflammatory 

agent. While immunostimulatory DNA 

sequences containing CpG motifs can have beneficial therapeutic 

effects, excessive stimulation of the innate immune 

system by bacterial and CpG DNA can cause detrimental 

effects, including inflammation, tissue damage and autoimmune 

diseases. 96-100 In fact, mammalian DNA, and specific 

sequences derived from mammalian DNA, have been 

shown to block activation of the immune system and 

decrease the production of proinflammatory cytokines 

caused by prokaryotic DNA or immunostimulatory DNA 

sequences with CpG motifs through an as yet uncharacterized 

mechanism(s). 96-99 Such antiinflammatory effects been 

demonstrated both in vitro and in an in vivo lung inflammation 

model. 99 As further evidence of the antiinflammatory 

effects of mammalian DNA, methylated calf thymus 

DNA was used to block immune activation by CpG DNA 

derived from the bacterium Escherichia coli. 101 

Antiinflammatory effects have also been demonstrated 

with exogenous nucleosides. Inosine, a purine nucleoside 

formed by the enzymatic deamination of adenosine, has 

been shown to have antiinflammatory activities in an in 

vivo murine model of acute lung inflammation induced by 

bacterial lipopolysaccharide. 102 Upon intratracheal instillation, 

inosine suppressed the production of proinflammatory 

cytokines such as IL-1-beta, IL-6 and TNF-alpha, while 

production of antiinflammatory cytokine IL-4 was 

enhanced. 102 Moreover, inosine had other antiinflammatory 

effects resulting in improved lung morphology, such as 

reduced polymorphonuclear neutrophil migration, edema, 

and nitric oxide production. 102 Inosine also had antiinflammatory 

effects on cultured human monocytes, neutrophils 

and epithelial cells, and it inhibited production of proinflammatory 

cytokines in vitro. 103-104 The mechanism of 

inosine action is unclear; one possibility suggested in the 

literature was signaling via A2 purinergic receptors. 102 

Interestingly, it was noted that while relatively large doses 

of inosine were required in the in vivo experiments 

described, a local route of administration might achieve 

desired anti-inflammatory effects at markedly lower 

doses. 102 

15

5. Decreased Mucus Viscosity and Improved 

Mucociliary Clearance via Increased Production of 

Antiinflammatory Cytokine IL-4 

In addition to antiinflammatory effects, IL-4 has been 

shown to decrease sodium absorption while increasing chloride 

secretion in cultured lung cells. 105 Such changes in ion 

transport in vivo could facilitate improved airway surface liquid 

properties, notably decreased mucus viscosity and 

increased mucociliary clearance. 105-106 Accordingly, it is 

hypothesized that if microdose DNA can increase the production 

of IL-4, this would both decrease inflammation and 

beneficially increase hydration of airway surfaces. These 

effects could be important in respiratory diseases such as cystic 

fibrosis, in which there is a need to decrease the viscosity 

of the mucus and/or improve mucociliary clearance. 

However, IL-4 is not only an important antiinflammatory 

cytokine, 107-109 but also a key cytokine involved in Th2-type 

humoral immune response. 110-112 Cytokine imbalances favoring 

a Th2 immune response have been implicated in allergic 

respiratory diseases such as asthma. 77-80, 111-112 As microdose 

DNA has in fact been successfully used for treating asthma 

and other allergic respiratory diseases, 26-31 it is speculated 

that enhanced IL-4 production by microdose DNA could help 

restore a favorable balance between airway surface fluid 

absorption and secretion within the respiratory tract. 

CLINICAL TRIALS OF MICRODOSE DNA (HP-3) 

Microdose DNA has been tested in three FDA-authorized, 

double-blind, placebo-controlled Phase II clinical trials 

for efficacy in the treatment of a) chronic bronchitis, b) 

COPD and c) CF. The chronic bronchitis trial involved 49 

eligible patients with the disease as defined by the American 

Thoracic Society. They were randomized to either micro- 

dose DNA (25 patients) or placebo (24 patients). After an 

initial 7-day placebo lead-in period, patients were treated 

with sublingually administered drops of either microdose 

DNA or placebo (vehicle control) daily for 90 days, with a 

subsequent evaluation one month after termination of treatment. 

Patients treated with microdose DNA showed a 

marked increase in sputum expectoration, clearing of the 

airways and significantly improved respiratory capacity. 

FEV1 (forced expiratory volume in one second, a measure 

of airflow rate) increased over 5%, and FEF25-75% (forced 

expiratory volume, a measure of small airway function) 

showed a statistically significant clinical improvement (p = 

0.007; Figure 1). There were no serious adverse events 

related to treatment with microdose DNA, and the type and 

frequency of reported side effects were comparable in the 

treatment and placebo groups. 

The COPD trial involved 48 eligible patients with the 

disease as defined by the American Thoracic Society. This 

study was conducted at a single site, with patients randomized 

to either microdose DNA (23 patients) or placebo (25 

patients). Patients were treated with sublingually administered 

drops of either microdose DNA or placebo daily for 

83 days subsequent to a 7-day placebo lead-in period. 

Initial and final evaluations involved a number of diseaserelated 

parameters. Significant improvement was seen in 

the blood oxygenation test (measured by pulse oximetry) 

and in the Distance Walked in 6 Minutes test (p = 0.019; 

Figure 2). Interestingly, many of the COPD patients had 

dry coughs with little or no sputum production; hence there 

was no increase in sputum output. As with the chronic bronchitis 

clinical trial, there were no serious adverse events 

related to treatment with microdose DNA. The type and 

frequency of side effects in the treatment and placebo 

groups were comparable. 

Figure 1. Mean change in baseline values (mean +/- standard error of the mean) in the FEF 25-75% (forced expiratory volume in the 

middle half of the testing range) in chronic bronchitis patients treated with either microdose DNA (square) or placebo (diamond). 


There were 37 patients (randomized to 17 treatment, 20 

placebo) enrolled in the CF clinical trial. All patients in this 

trial had either mild or moderate disease. Patients were 

treated for 8 weeks with sublingual drops of microdose 

DNA or placebo. Evaluations were conducted at 1, 4 and 8 

weeks after treatment was initialized. The main parameters 

evaluated were FEV 1, the FEV 1/FVC (forced expiration 

volume/forced vital capacity) ratio, a measure of airway 

obstruction, and the FEF 25-75%. Patients treated with micro- 


dose DNA showed a trend toward clinical improvement in 

all 3 parameters (Figures 3A-3C). Statistical significance 

was not achieved owing to both the small number of subjects 

and the fact that this study was limited to patients with 

mild or moderate CF. However, these results, coupled with 

observed subjective increases in sputum clearance, underscored 

the potential of microdose DNA in improving pulmonary 

function in CF patients. 

Figure 2. Mean change from baseline values (mean +/- standard error of the mean) in the Distance Walked in 6 Minutes 

test of COPD patients treated with either microdose DNA (square) or placebo (diamond). 

Figure 3A. Mean percent change from baseline values (mean +/- standard error of the mean) in pulmonary function tests 

of cystic fibrosis patients treated with either microdose DNA (square) or placebo (diamond). FEV 1= forced expiratory volume 

in one second. 

17

Figure 3B. Mean percent change from baseline values (mean +/- standard error of the mean) in pulmonary function tests 

of cystic fibrosis patients treated with either microdose DNA (square) or placebo (diamond). FEV 1/FVC = forced expiration 

volume/forced vital capacity ratio. 

Figure 3C. Mean percent change from baseline values (mean +/- standard error of the mean) in pulmonary function tests 

of cystic fibrosis patients treated with either microdose DNA (square) or placebo (diamond). FEF 25-75% = forced expiratory 

volume. 

EVIDENCE-BASED CLINICAL EXPERIENCES 

WITH MICRODOSE DNA 

In addition to the clinical trials described above, the 

utility of microdose DNA in alleviating the symptoms of 

various respiratory diseases, as well as that of otitis media, 

has been demonstrated through evidence-based clinical testing. 

Examples of these clinical experiences, extracted from 

available patent literature, have been summarized in a separate 

publication. 32 The results of evidence-based testing 

suggest that microdose DNA has broad potential utility in 

relieving symptoms of a variety of respiratory ailments, 

including CF, COPD, chronic bronchitis, asthma, respirato- 

18 

ry allergies, radiation-induced mucositis, respiratory disease 

resulting from occupational/environmental chemical 

exposure, chronic upper respiratory illness, respiratory 

congestion and otitis media. 32 Moreover, these results, 

along with the Phase II clinical trial data, indicate that 

microdose DNA has a good safety profile. It also appears 

that the product can be administered safely in conjunction 

with other medications. 

Additionally, there have been numerous anecdotal 

reports that immediately upon the onset of cold symptoms, 

sublingual administration of one or two drops of microdose 

DNA 4-8 times daily for 5-10 days can alleviate the dura- 


tion and/or severity of cold symptoms such as congestion, 

excessive nasal discharge and sore throat. (Some individuals 

administer as many as four sublingual drops per hour at 

cold onset and then reduce that number over the next several 

days). If meaningful reductions in the duration and severity 

of common cold symptoms can be demonstrated through 

formal clinical investigations, microdose DNA may have 

utility as a favored treatment for cold symptoms. 

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Phytonutrients May Decrease Obstetric 

Complications: A Retrospective Study 

Presented at the 70th Annual Meeting of the Central Association of 

Obstetricians and Gynecologists, October 2003, San Diego, California 

C. Doug Odom, MD, 1 Suneet P. Chauhan, MD, 2* Everett F. Magann, MD, 2** Rick W. Martin, MD, 2 

Carl H. Rose, MD, 2*** John C. Morrison, MD 2++ 

1. Department of Obstetrics and Gynecology, Mississippi Baptist Medical Center, 

Jackson, Mississippi (CDO) 

2. University of Mississippi Medical Center, Jackson, Mississippi 

++ Correspondence: 

John C. Morrison, MD 

Department of Obstetrics & Gynecology 

University of Mississippi Medical Center 

2500 North State Street 

Jackson, Mississippi 39216-4505 

Phone: 601-984-5350 Fax: 601-984-5378 

E-mail: jmorrison@ob-gyn.umsmed.edu 

*Present address: Aurora Women’s Pavilion, West Allis, Wisconsin 

**Present address: Naval Medical Center, Portsmouth, Virginia 

***Present address: Mayo Clinic, Rochester, New York 

ABSTRACT 

Objective 

Comparison of post-delivery pregnancy outcome variables 

in women who either did or did not add to their daily 

diet a concentrated, encapsulated fruit and vegetable juice 

powder (FVJP) along with standard prenatal vitamins during 

gestation. 

Design 

Retrospective descriptive analytic comparison of charted 

singleton pregnancy outcome variables at delivery. 

Setting 

Private obstetrics and gynecology practice in Jackson, 

Mississippi. 

Subjects 

The pregnancy outcome information was collected 

from 356 pregnant women, half of whom added FVJP to 

their standard prenatal care. 

Measures of Outcome 

Documented obstetric complications by medical staff 

at the time of delivery. 

Results 

Women who added FVJP to their prenatal regimen had 

fewer Cesarean deliveries (47% vs 66%), no delivery 

before 37 weeks (0 vs 20%), and no diagnosis of 

preeclampsia (0 vs 21%) compared to women who did not. 

Conclusions 

The findings of this retrospective chart review suggest 

that adding nutrition from FVJP to standard prenatal vitamin 

use may reduce the incidence of some obstetric complications. 

More study is needed to confirm these observations. 

INTRODUCTION 

Current nutritional recommendations for pregnant women 

are based on the USDA Food Guide Pyramid and include 

four servings of vegetables and three servings of fruit, 

daily. 1 However, only 20.6% of women surveyed in 

23

Mississippi (MS) reported consuming five or more servings 

of fruits and vegetables per day in 2003. These foods, particularly 

the deeply pigmented varieties, provide a wide 

array of phytonutrients with antioxidant activity, including 

carotenoids and vitamins such as folate and vitamin C, 

while also being low in calories and fat. 3 As recently 

reviewed by Roberts et al, nutritional interventions anticipated 

to reduce complications during pregnancy have 

focused mainly on macronutrients such as protein or specific 

lipids, minerals such as calcium, zinc or iron, and relatively 

high doses of isolated vitamins such as vitamin C and 

vitamin E. 4 Although oxidative stress is one of several etiologies 

theorized to be involved in the development of 

preeclampsia, currently there are no studies available on the 

incidence of this complication in the final trimester and at 

delivery that focus on increased fruit and vegetable consumption 

in pregnant women throughout gestation. 

Common and undesirable obstetric complications 

include preterm labor, preterm birth, preterm premature 

rupture of membranes (PPROM), low birth weight, 

preeclampsia, and intrauterine growth restriction. This retrospective 

descriptive study is from one OBGYN practice 

averaging 471 deliveries annually in Jackson, Mississippi, 

a practice that includes many women at high risk for obstetric 

complications. The study was initiated after the nurses 

noticed that mothers who used a specific fruit and juice 

powder product (FVJP) reported anecdotally that they experienced 

fewer obstetric complications than other mothers 

served by the same practice. This nutritional product has 

been reported to both increase the concentration of blood 

antioxidants5-7 and reduce homocysteine, 8,9 an amino acid 

reported elevated in women who developed preeclampsia. 4 

These studies in healthy adults provided a biologic rationale 

for the observations of the nursing staff. The purpose of this 

investigation was to evaluate the hospital records for docu- 

Table 1. Socioeconomic and demographic characteristics of the groups 

mented complications at delivery of mothers from this 

OBGYN practice who either chose to use a specific nutritional 

FVJP supplement (Group I) or not (Group II). 

MATERIALS AND METHODS 

Subjects 

The study was approved by the Human Use Committee 

of Mississippi Baptist Medical Center, Jackson, 

Mississippi, for retrospective chart review. This hospital is 

one of four in the Jackson area accepting obstetric patients. 

We identified 178 women within the practice who reported 

taking the FVJP between January 1, 2000, and December 

31, 2002, as Group I. The 178 comparison women making 

up Group II were selected as the next consecutive deliveries 

within the practice group who did not report taking 

FVJP and matched the Group I women by age (within 3 

years), parity (+1), ethnicity, prior preterm birth history 

(+1) and private insurance coverage (yes or no). The groups 

were not matched for marital status and previous obstetric 

or medical history other than preterm birth. A total of 356 

singleton pregnancies were included. 

The hospital records were reviewed by a technician and 

obstetric complications were tabulated. A physician verified 

the accuracy of the data entered. For the purpose of this 

analysis: preterm labor was defined as labor before 37 

weeks requiring medication to control; preeclampsia was 

documented by the attending physician; preterm premature 

rupture of membranes was defined as spontaneous ruptured 

membranes prior to 37 weeks gestation; and fetal distress 

was defined by late decelerations or non-reassuring nonstress-test 

as assessed by the attending physician. 

The composition of the retrospective study groups is 

summarized in Table 1. During the 3-year observation peri- 

Parameter Group I Group II 

(n=178) (n= 178) 

Age, years 

Race 

28.7 + 5.2 28.0 + 5.1 

Caucasian 152 158 

African American 18 18 

Other 8 2 

Nulliparous 77 (43%) 82 (46%) 

Covered by private insurance 165 (93%) 164 (92%) 

Married 

Previous obstetric/medical history 

166 (93%) 161 (90%) 

Preterm delivery 10 12 

Preterm premature rupture of membranes 0 1 

Preterm premature rupture of preeclampsia 16 9 

Diabetes (idiopathic) 

Chronic hypertension 

3 

3 

2 

2 


Table 2. Pregnancy outcomes 

od, 83% (147 of the 178) of Group I women began taking the 

FVJP before 13 weeks gestation and all women in this group 

were taking it by 28 weeks gestation. The FVJP capsules 

(Juice Plus+ ®, NSA, Inc., Memphis, TN) contain primarily 

fruit (apple, orange, pineapple, cranberry, peach, acerola 

cherry, papaya) and vegetable (carrot, parsley, beet, kale, 

broccoli, cabbage, spinach, tomato) juice concentrate powder. 

Four capsules daily provide several nutrients common 

in plant foods (phytonutrients), including approximately: 

beta-carotene equivalent to 12,500 IU of vitamin A activity; 

234 mg vitamin C; 45 IU vitamin E as alpha-tocopherol; 420 

mcg folate; 60 mg calcium; and about 10 calories. 

STATISTICAL ANALYSIS 

Kolmogorov-Smirnov test was used to determine if the 

data followed Gaussian distribution, and Student t-test or 

Mann-Whitney tests were used where applicable (GraphPad 

InStat version 2, GraphPad Software, Inc., San Diego, 

California). Odds ratio (OR) and 95% confidence intervals 

(CI) were calculated. If there was a zero in one of the 2x2 

contingency cells, then 0.5 was added to each value and 

approximation of Woolf was used. A P value less than 0.05 

was considered significant. 

RESULTS 

The matching resulted in study groups that were not different, 

as shown in Table 1, including for unmatched variables 

such as marital status. Undesired obstetric outcomes in 


Delivery Parameters Group I Group II P value 

(n=178) (n=178) OR (95% CI) 

Average gestational age at delivery 39.3 + 0.9 38.2 + 2.4 > 0.0001 

Less than or equal to 32 weeks 0 8 (4%) 17.8 (1.0, 311) 

Less than 37 weeks 0 35 (20%) 88.3 (5.4, 1453.2) 

Greater than or equal to 41 weeks 9 (5%) 10 (6%) 1.1 (0.4, 2.8) 

Preterm delivery secondary to: 

Preeclampsia 0 38 (21%) 97.8 (5.95, 1607.4) 

Spontaneous labor 0 16 (9%) 0.03 (0.0, 0.5) 

PPROM 0 6 (3%) 0.14 (0.0,2.7) 

Total Cesarean deliveries 83 (47%) 117 (66%) 2.2 (1.4, 3.4) 

secondary to: 

Cephal pelvic disproportion 78 (44%) 85 (48%) 0.9 (0.6, 1.3) 

Fetal distress 3 (2%) 11 (6%) 2.5 (0.1, 0.9) 

Breech 3 (2%) 2 (1%) 1.5 (0.3, 9.1) 

Herpes simplex virus 3 (2%) 2 (1%) 1.5 (0.3, 9.1) 

Preeclampsia 0 12 (7%) 0.04 (0.002, 0.6) 

Group I and Group II mothers are listed in Table 2. 

Gestational age averaged one week longer in Group I 

women than Group II women, and no Group I woman delivered 

before 37 weeks gestation, compared to 46 of the 

Group II women. No Group I woman had preterm delivery, 

while 44 Group II women had preterm delivery (38 secondary 

to preeclampsia). The frequency of those delivering 

post-term (greater than 41 weeks) was similar between the 

two groups. Cesarean deliveries were performed on women 

in both groups (83 in Group I, 117 in Group II), although 

analysis shows a protective effect in Group I (OR 2.2). No 

Group I woman had a Cesarean delivery due to preeclampsia, 

compared to 12 in Group II. Cephal pelvic disproportion, 

breech, and infection (Herpes simplex virus) were similar 

in their occurrence between the two groups. 

Various neonatal factors differed between the two 

groups (Table 3). Average birth weight was significantly 

higher (P=0.0003) among Group I (3,507 + 424 grams) 

compared to Group II (3,280 + 709 grams). Babies born to 

Group I mothers had a lower frequency of neonatal intensive 

care unit (NICU) admission (0 vs.17) when compared 

to Group II offspring (OR 38.7 CI 2.3, 648.9). The majority 

of these neonatal intensive care unit (NICU) admissions 

(13) in Group II were due to respiratory distress syndrome. 

DISCUSSION 

These retrospective observations support the hypothesis 

that the women who chose to take FVJP capsules during 

pregnancy in this practice and during this time frame, car- 

25

Table 3. Neonatal outcomes 

ried their babies longer than 37 weeks, had fewer babies 

weighing less than 2,500 grams, did not have preterm labor 

requiring intervention and did not require elective delivery 

due to preeclampsia. Although every attempt was made to 

assure the chart review was thorough and unbiased, the 

Group I women may have differed from the Group II 

women independent of the nutritional supplement in a way 

not apparent from the charted information. Pregnant women 

are advised to consume at least seven servings of fruits and 

vegetables every day when they are expecting, 1 yet very few 

women in Mississippi even consume five daily servings. 2 A 

study conducted in second trimester pregnant women in 

North Carolina found that higher-income, older, and better 

educated women reported consuming 3-5 servings of vegetables 

per day. 10 This retrospective study matched for age 

and insurance coverage, but it is possible that Group I 

included mothers whose higher educational status was 

higher than that of Group II mothers. 

While it is not appropriate to assume that any nutritional 

product can replace all the components found in produce, 

it is possible to consider that the FVJP capsules may 

have complemented the nutritional status of the Group I 

women. Oxidative stress has been linked during pregnancy 

to preterm labor as well as low birth weight, preterm premature 

rupture of membranes (PPROM), preeclampsia, 

intrauterine growth restriction (IUGR), and numerous newborn 

complications. Endothelial dysfunction has been theorized 

to contribute to development of preeclampsia. It has 

previously been reported that healthy, adult study subjects 

given these FVJP capsules showed reduction in several indicators 

of oxidative stress in the body, specifically increased 

plasma ferric reducing/antioxidant power (FRAP) 8 and 

reduced lipid peroxidation. 6,7 It has also been reported to 

maintain normal vasoactivity in humans after a high-fat test 

meal. 11 While these investigations were not performed on 

pregnant subjects, it is reasonable to expect similar functional 

findings that might explain the reduction in frequency of 

undesirable obstetric outcomes noted in the Group I mothers. 

26 

Infant at birth Group I Group II P value 

(n=178) (n=178) OR (95% CI) 

Average birth weight (grams) 3,507 + 424 3,280 + 709 0.0003 

< 1,500 0 2 (3%) 7.1 (0.4, 139) 

< 2,500 2 (1%) 22 (12%) 12.4 (2.9, 53.6) 

< 4,000 20 (11%) 20 (11%) 1.0 (0.5, 1.9) 

< 4,500 4 (2%) 1 (0.5%) 4.1 (0.5, 36.8) 

NICU Admission 0 17 (10%) 38.7 (2.3, 648.9) 

Respiratory Distress Syndrome 0 13 (8%) 29.1 (1.7, 494.1) 

Findings from several recent studies illuminate the 

observations from this retrospective. Women in the lowest 

tenth percentile for preconception vitamin C intake were at 

greater risk for preterm delivery and PPROM, although the 

odds were slightly improved when second trimester intake 

was higher than intake before pregnancy. 12 Another study 

found both placental tissue and maternal blood had lower 

carotenoid concentrations from mothers with preeclampsia 

than from those without. 13 Lower levels of tocopherols have 

been reported in women with preeclampsia compared to 

mothers without. 14 Blood from Italian mothers with a diet 

poor in fruits and vegetables showed a decrease each 

sequential trimester in total antioxidant capacity, with 

umbilical cord blood values correlated to the maternal values 

at delivery. These findings led the authors to conclude 

that “efforts should be made to improve dietary habits in 

pregnancy.” 15 These and other nutrients are all present in 

the FVJP capsules used by the Group I women. Daily consumption 

of antioxidant nutrients in the FVJP capsules may 

be responsible for the significantly decreased rate of complications 

noted in these 178 deliveries, such as preterm 

labor, birth before 37 weeks, preeclampsia, NICU admission 

and infant respiratory distress syndrome. If these findings 

are confirmed in a randomized, prospective clinical 

trial currently being conducted, a simple and inexpensive 

nutritional solution may be available to effectively address 

common and costly obstetric complications. 

ACKNOWLEDGMENT 

No corporate funding was provided to support this 

study. One author, C. Doug Odom, MD, has an individual 

financial interest in the FVJP that was used by patients in 

the study. None of the other authors have any financial 

interests in the FVJP used by patients in the study. 


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J Hum Nutr and Diet. 2000;13:21-27. 

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plasma carotenoid, alpha-tocopherol, and lipid peroxide 

levels in response to supplementation with concentrated 

fruit and vegetable extracts: a pilot study. Curr Ther Res. 

1996; 57:445-461. 

8. Samman S, Sivarajah G, Man JC, Ahmad ZI, Petocz P, 

Caterson ID. A mixed fruit and vegetable concentrate 

increases plasma antioxidant vitamins and folate and 

lowers plasma homocysteine in men. J Nutr. 

2003;133:2188-2193. 

9. Panunzio MF, Pisano A, Antoniciello A, Di Martino V, 

Frisoli L, Cipriani V, Mongelli MA, Bronzetti G. 

Supplementation with fruit and vegetable concentrate 

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trial. Nutr Res. 2003;23:1221-1228. 

10 Bodnar LM, Siega-Riz AM. A diet quality index for 

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27


Pilot Study: Flaxseed Supplementation Was 

Effective in Lowering Serum Glucose and 

Triacylglycerol in Glucose Intolerant People 


Y Rhee, PhD 

Department of Health, Nutrition and Exercise Sciences 

North Dakota State University, 351 EML 

Fargo, ND, 58105. 

Phone: 701-231-7476 Fax: 701-231-7174 

E-mail: yeong.rhee@ndsu.edu. 

Yeong Rhee, PhD,* Ardith Brunt, PhD 

Department of Health, Nutrition and Exercise Sciences 

North Dakota State University, Fargo, North Dakota 

ABSTRACT 

Background: Glucose intolerance increases the risks 

of diabetes and cardiovascular disease development. 

Objective: The effects of full-fat yellow omega flaxseed 

(flaxseed) or durum wheat bran (wheat bran) supplementation 

on fasting serum glucose (FSG), total cholesterol (TC), 

triacylglycerol (TG), high-density lipoprotein (HDL) cholesterol 

and low-density lipoprotein (LDL) cholesterol concentrations 

in glucose intolerant people were investigated. 

Design: To screen subjects for glucose intolerance, an 

oral glucose tolerance test was performed. Nine glucose intolerant 

people received either 40 g of flaxseed or wheat bran for 

12 weeks, followed by a 4-week washout period. Then they 

received the other supplement for the next 12 weeks. 

Results: Baseline dietary intake, FSG, TC, TG, HDL, 

and LDL among subjects was not different. No differences 

in weight, TC, HDL, and LDL between treatment groups or 

in individuals were observed 12 weeks later. However, 12 

weeks of flaxseed supplementation significantly lowered 

FSG compared to baseline (p=0.02) and wheat bran supple- 

mentation (p=0.04). Twelve weeks of flaxseed supplementation 

significantly lowered TG compared to wheat bran 

supplementation (p=0.02). Flaxseed supplementation 

showed a superior effect on lowering FSG and TG when 

compared to wheat bran supplementation. 

Conclusion: These results suggested that flaxseed may 

lower FSG and TG in glucose intolerant individuals. 

Key Words: glucose intolerance, flaxseed, wheat bran, 

antioxidant, fasting serum glucose, triacylglycerol. 

INTRODUCTION 

The number of people with diabetes in the US has been 

increasing steadily in all age groups over the past ten years, 

with larger increases in younger populations. For example, 

the incidence of type 2 diabetes increased 70% among people 

aged 30 to 39 years in the past ten years. 1,2 In 2002, 

18.2 million people were diagnosed with diabetes, about 

90-95% of them with type 2 diabetes. 3 About 15.6% of the 

population aged 40-74 years have impaired glucose tolerance. 

4 Individuals with impaired glucose tolerance have an 

increased risk of developing type 2 diabetes and cardiovascular 

disease. 4 About 2.3-11% (5-8% average) of people 

with impaired glucose tolerance develop type 2 diabetes 

annually. 5 

Increasing evidence now indicates that antioxidants 

play roles in the prevention and/or delay of diabetes and the 

development of diabetic complications. 6-8 Reactive oxygen 

species (ROS) and increased oxidative stress have been 

implicated in the development of diabetes and diabetic 


complications such as microvascular disease or macrovascular 

disease. 9-11 Hyperglycemia found in those with impaired 

glucose tolerance and diabetes generates ROS. 9,10,12 ROS 

generation leads to increased oxidative stress as indicated by 

lipid peroxidation or oxidative DNA damage, 13-15 impaired 

ROS scavenging effects of antioxidants, 10 and decreased 

antioxidant concentrations in diabetics. 11 Increased oxidative 

stress by ROS leads to cell damage or cell death in various 

tissues, including pancreatic beta-cells. 6 Pancreatic 

beta-cell damage or death further impairs glycemic control 

in people with impaired glucose tolerance or diabetes. 6 

Thus, prolonged poor glycemic control may result in the 

development of diabetes and diabetic complications. 

Several studies showed that flaxseed has antioxidant 

activity due to secoisolariciresinol diglucoside (SDG), a 

plant lignan, and its metabolites secoisolariciresinol 

(SECO), enterodiol (ED), and enterolactone (EL) which are 

all formed in the body. 16-18 Prasad17,19 reported that flaxseed 

supplementation aids in the prevention or delay of type 1 

and type 2 diabetes development in diabetes-prone animals. 

SDG, ED, and EL inhibited, in vitro, DNA scissions and 

linoleic acid peroxidation, which also indicate antioxidant 

activity of lignan and its metabolites. 20 In addition, SDG 

supplementation was associated with decreased serum or 

pancreatic malondialdehyde (MDA), which is a lipid peroxidation 

product, and decreased ROS-producing activity of 

white blood cells (WBC) (chemiluminescent activity of 

WBC, WBC-CL). Both MDA and chemiluminescent activity 

of WBC are used as indicators of oxidative stress and 

decreased antioxidant concentrations in the body. 17,19 

Comparing antioxidant activity of SDG, SECO, ED, and EL 

to that of vitamin E, antioxidant activities were found to be 

1.27, 4.86, 5.02, and 4.35 times as potent as vitamin E for 

SDG, SECO, ED, and EL, respectively. 18 Moreover, whole 

or defatted flaxseed or SDG supplementation showed lipidlowering 

effects in animals and humans. 21-24 Limited studies 

have reported dietary flaxseed supplementation effects 

on cardiovascular disease and especially, diabetes development 

in humans. Thus, the objective of this study was to 

determine if 12 weeks (3 months) of supplementation with 

40 g/day of full-fat yellow omega flaxseed (flaxseed) or 40 

g/day of durum wheat bran (wheat bran) will decrease indicators 

of type 2 diabetes and cardiovascular disease in individuals 

with impaired glucose tolerance. 

The research protocol was reviewed and approved by the 

Institutional Review Board at North Dakota State University. 

Written informed consent was obtained from all participants 

before the screening test and initiation of the study. 

MATERIALS AND METHODS 

Subject Selection 

Individuals at high risk for developing type 2 diabetes–overweight, 

with high blood pressure and a family 


history of diabetes–were targeted for the initial screening. 

Subjects were screened initially for impaired glucose tolerance 

using a 12-hour fasting blood sample. To be eligible 

for this study, an individual was required to have fasting 

plasma glucose between 100-125 mg/dl, and following a 

100 g oral glucose load, have 2-hour plasma glucose of 

more than 140 mg/dl but less than 199 mg/dl. At the 

screening, subjects filled out a health questionnaire. 

Individuals were excluded from this study if they were taking 

oral hypoglycemic medication or insulin injection. In 

addition, an individual who had any diagnosed illness other 

than controlled hypertension or impaired glucose tolerance 

or who was allergic to either flaxseed or wheat was excluded 

from the screening test. There were no restrictions in 

regard to race or socioeconomic status. A total of 33 people 

were screened and 11 people were qualified for study 

participation. 

Design and Treatment Groups 

A randomized crossover research design was used for the 

study. Eleven subjects were randomly assigned into one of 

two groups. Subjects received a daily allotment of either 40 g 

of wheat bran or flaxseed in the form of ground grain or bread 

for 12 weeks. They were to incorporate the supplement in 

their daily meals using a method of their choice. Following a 

4-week washout period, they received the alternate supplement 

(either flaxseed or wheat bran) for another 12 weeks. 

The ingredients in the bread were adjusted to provide 

similar calories and fat content in both supplemental breads. 

A local bakery prepared loaves of bread for the study. 

Before entering the study, participants were instructed to 

substitute the study supplement for some of their usual food 

consumed at the meal, so that no more calories than usual 

would be consumed. Participants were requested to keep the 

loaves of bread and/or ground supplement in the freezer to 

maintain freshness. Participants were also asked to record 

the amount eaten and to return leftover supplements to the 

investigators. The collected leftover supplements were 

weighed by the investigators to check compliance. 

Proximate Analysis of Supplements 

Ground full-fat yellow omega flaxseed and durum wheat 

bran were provided by the North Dakota Oilseed (flaxseed) 

Council, Bismark, ND, and Dakota Growers Pasta Co., 

Carrington, ND, respectively, and used as supplements in this 

study. The proximate analysis of yellow omega flaxseed and 

durum wheat bran was performed by Medallion Labs, 

Minneapolis, by Drs. Hammond, Manthey and Vick at North 

Dakota State University. Dr. Wiesenborn at North Dakota 

State University analyzed flaxseed for lignan content. 25 

Nutrient Analysis of Supplemental Bread 

Nutrient compositions of wheat bran bread or flaxseed 

bread were analyzed using Diet Analysis Plus software 

(version 6.0, ESHA Research, Salem, OR). 

29

Health Questionnaire 

A health questionnaire that included birth date, current 

health conditions, medication use, vitamin, mineral, or herbal 

supplement use, and exercise practices was completed at the 

screening. A follow-up health questionnaire was completed 

at the end of the study to determine if any changes had 

occurred in the subject's health or exercise practices. 

Anthropometric Measurement 

Each subject's height (without shoes) was measured 

during the screening test. Using a balance beam scale, each 

subject was weighed at the screening and at the beginning 

and end of each supplementation. Body mass index (BMI) 

was calculated using the equation BMI = weight 

(kg)/height2 (m2). 26 The percent body fat was measured 

using a bioelectrical impedance analyzer (Bodystat 

1500MDD, Bodystat Ltd., Douglas, British Isles); waist and 

hip circumferences were measured using a measuring tape. 

Dietary Intake 

The Block Brief 2000 Food Questionnaire, a selfadministered 

66-item semi-quantitative food frequency 

questionnaire, was administered at the beginning of each 

supplementation. Compared to multiple 24-hour recalls, 

this semi-quantitative food frequency questionnaire 

assessed dietary intake more accurately. 27 This questionnaire 

assesses intake over the past year; therefore, measuring 

dietary intake more than twice in a year would not 

improve assessment of past dietary intake. 

Blood Sample Collection 

After a 12-hour fast, subjects had blood samples drawn 

a total of four times. Samples were collected before and 

after each 12-week supplementation. Fasting serum glucose 

(FSG), total cholesterol (TC), triacylglycerol (TG), 

low-density lipoprotein (LDL) cholesterol, and high-density 

lipoprotein (HDL) cholesterol concentrations were measured 

by MeritCare Health System Laboratory, Fargo, ND, 

by an automated chemistry analyzer. 

STATISTICAL ANALYSIS 

Data Analysis 

Changes in the dependent variables after 12 weeks of 

supplementation were compared within the treatment group 

and between treatment groups using SAS software (version 

9.1; SAS, Cary, NC). Data were tested for normality and 

equality of variances (Univariate test). Comparisons of 

changes in variables between treatment groups were tested 

by two sample t-tests. Comparisons of changes in variables 

within the treatment group were tested by paired t-tests. 

Significance level was set at p < 0.05. All data are reported 

as mean ± SD. 

Dietary Analysis 

To evaluate usual intake over the past year, the Block 

Brief 2000 Food Questionnaire was computed. Changes in 

caloric, macronutrient, vitamin and mineral intake were 

analyzed using SAS software (version 9.1; SAS, Cary, NC). 

A generalized linear model (GLM) was used for analysis. 

RESULTS 

A total of eleven subjects initially participated in the 

study. Four participants dropped out: two after first blood 

sample collection and another two after the third. The first 

two subjects withdrew from the study due to personal reasons 

unrelated to the study; the latter two subjects dropped 

out of the study because they did not like the wheat bran 

supplement. The two who dropped out during the first 

phase of supplementation were excluded from the data 

analysis; however, the subjects who dropped out during the 

second phase of supplementation were included in the data 

analysis. The overall compliance rate with dietary supplements 

was > 95% in these subjects. 

Proximate Analysis of Supplements 

Forty grams of flaxseed provided 640 mg of total lignan, 

17 g of fat (53.3% alpha linolenic acid), and 9.8 g (24.6%) of 

fiber. Forty grams of wheat bran provided no lignan, 1.9 g of 

fat (3.9% alpha linolenic acid), and 16.2 g (40.5%) of fiber. 

Nutrient Analysis of Wheat Bran or Flaxseed Bread 

Supplemental bread that contained 40g of either 

ground flaxseed or wheat bran, provided 453 kcal and 398 

kcal, respectively. Table 1 shows the nutrient composition 

of supplemental bread. Software limitations prevented 

analysis of alpha linolenic acid and insoluble and soluble 

fiber content in the flaxseed or wheat bran bread. 

Characteristics of Subjects 

The average age of the participants was 54.67 ± 6.61 

years with a range of 50-66 years. Of the nine participants, 

four were male and five were female. Subjects reported no 

change in dietary intake, health status, or exercise habits 

throughout the study. All participants were nonsmokers 

TABLE 1. Composition of daily supplemental flaxseed 

bread and wheat bran bread. 

Measures Flaxseed Wheat bran 

Energy (kcal) 453 398 

Carbohydrate (g) 72.2 83.4 

Fiber (g) 13.1 18.6 

Protein (g) 16.8 14.8 

Fat (g) 16.4 6.01 

Saturated fatty acids (g) 6.65 0.82 

Monounsaturated fatty acids (g) 4.50 1.25 

Polyunsaturated fatty acids (g) 4.96 3.20 


and reported exercising for 30 minutes or more per day, 3-5 

days per week. Six participants reported taking multivitamins; 

two participants reported taking calcium/magnesium 

supplements; and one participant reported taking a vitamin 

E supplement. 

Anthropometrics 

Weight change and body mass index (Table 2) were not 

significantly different compared to the baseline, nor between 

treatment groups. The waist-hip ratio decreased after 12 

weeks of wheat bran supplementation compared to the baseline 

(p

TABLE 4. Fasting blood lipid concentrations over the course of the experiment. 1 

flaxseed or wheat bran supplementation, respectively; however, 

these changes were not significantly different (Table 4). 

DISCUSSION 

Fasting serum glucose concentration decreased significantly 

following 12 weeks of flaxseed supplementation. 

Although subjects reported no changes in dietary or exercise 

habits, flaxseed supplementation resulted in a 4 kg weight 

loss. Studies with a similar amount of weight loss (3.5 ± 5.5 

kg or 4.0 kg) resulted in a small (2 ±12 mg/dl) or no change 

in blood glucose concentration in people with impaired glucose 

tolerance. 28,29 These studies suggest that the weight 

loss in the current study would have a minimal or no effect 

on fasting serum glucose concentration in these subjects. 

Other investigators found similar glucose lowering 

effects of SDG supplementation in animal models. 17,19,30 

SDG supplementation delayed or prevented development of 

diabetes by 71-75%, and 80% in type 1 and type 2 diabetic 

animals, respectively. 17,19,30 These studies indicated that 

SDG supplementation decreased oxidative stress as indicated 

by decreased serum and pancreatic MDA and WBC-CL, 

and increased antioxidant reserves. These changes in oxidative 

stress and antioxidant reserves led to a decrease in type 

1 and type 2 diabetes development. 17,19,31 The delayed or 

prevented diabetes development by antioxidant supplementation 

in animal models has also been reported by other 

investigators. 32,33 These studies show that decreased fasting 

serum glucose concentration in the current study might be 

due to the antioxidant activity of SDG in the flaxseed rather 

than any other of its components. 

In addition to the glucose lowering effects of the 

flaxseed supplementation, several investigators have studied 

the effects of flaxseed supplementation on blood lipids related 

to the risk of cardiovascular disease development. 

Flaxseed supplementation successfully reduced the development 

of atherosclerosis by 46% and plaque formation in an 

animal model. 34,35 Moreover, flaxseed meal supplementation 

in the diet (20% of diet, 17.0 µg SDG/g flaxseed meal) 

significantly lowered TC and TG concentrations in rats with 

32 

Flaxseed Wheat Bran 

Baseline After 12 weeks Baseline After 12 weeks 

Total cholesterol (mg/dl) 211.9 ± 20.5 211.6 ± 26.4 226.2 ± 24.2 230.1 ±20.9 

TG (mg/dl) 201.6 ± 68.5 150.2 ± 63.6 2 161.9 ± 67.7 195.7 ± 86.8 

HDL (mg/dl) 47.1 ± 4.8 51.4 ±10.3 48.7 ± 8.3 52.4 ± 6.9 3 

LDL (mg/dl) 117.9 ± 19.1 123.3 ± 22.8 136.9 ± 22.9 128.7 ± 22.4 

1All values are mean ± SD; n=7-9. There were no significant differences in total cholesterol and LDL over the course of the 

experiment. 2Significantly decreased compared to 12 weeks of wheat bran supplementation, p=0.02. 3Significantly increased 

compared to the baseline, p=0.02. 

FIGURE 1. Fasting serum glucose concentrations before 

and after each supplementation. 

*Significantly decreased compared to the baseline, p=0.02, and 

compared to 12 weeks of wheat bran supplementation, p=0.04. 

normal lipid concentrations and in obese rats with high TC 

and TG. 36,37 Whole flaxseed supplementation also lowered 

LDL, lipoprotein (a), apolipoprotein A-I, and apolipoprotein 

B without altering other blood lipids in humans. 38,39 

Moreover, Jenkins and colleagues 22 showed that partially 

defatted flaxseed was effective in lowering TC, LDL and 

lipoprotein (a), a strong predictor of cardiovascular disease, 

with no adverse effects on HDL in humans. 

Supplementation of SDG, a lignan complex isolated from 

flaxseed, or Crop Development Center (CDC) flaxseed with 

very low alpha linolenic acid also lowered serum TC, LDL, 

TC/HDL, MDA, and aortic MDA in animal models. 21,23,24 

These studies indicate that the omega 3 fatty acid was not 

responsible for the hypocholesterolemic response; it might 

be more related to antioxidant activity of flaxseed lignan. 

The results from the current study showed similar trends 

compared to studies discussed above. Although it was not 

significantly different (p=0.11), 12 weeks of flaxseed supplementation 

resulted in a trend towards lowered TG concentrations 

by 25% and increased HDL concentrations by 

9% compared to baseline without affecting TC concentrations. 

Statistically non-significant data might be due to the 

small sample size in the current study. 


In the current study, changes in the FSG, TG, and HDL 

concentrations would be attributed to antioxidants in the 

flaxseed. Flaxseed supplementation provided a significant 

amount of antioxidants compared to wheat bran supplementation. 

Forty grams of flaxseed provided 640 mg lignan, 

which is known as the antioxidant in flaxseed. 

However, there are very limited amounts of antioxidants in 

wheat bran; its insoluble fiber contains ~0.5-1.0% phenolic 

groups which are known as antioxidants in whole grains. 40 

The antioxidants in flaxseed may have functioned to lower 

FSG and TG concentrations in these glucose intolerant subjects. 

The results from the current study and studies by 

other investigators suggest that flaxseed has superior antioxidant 

activity in lowering risks of diabetes and cardiovascular 

disease development. 

Daily fiber intakes in these subjects were not significantly 

different within the treatment group and between the 

treatment groups. One hundred grams of wheat bran and 

flaxseed contained about 37.9% and 17.9% insoluble fiber 

and 2.6% and 6.7% soluble fiber, respectively. Soluble fiber 

is known to have serum glucose lowering effects due to gel 

formation properties. The limited amount of soluble fiber in 

the wheat bran and in flaxseed supplement (1.04 g soluble 

fiber/40 g wheat bran vs. 2.68 g soluble fiber/40 g flaxseed) 

would not have caused any FSG or TG lowering effects in 

these subjects. Therefore, wheat bran would likely not have 

any effects or have fewer effects on serum glucose or lipid 

concentrations due to a very limited amount of soluble fiber 

and no antioxidant content. 

Forty grams of flaxseed and wheat bran provided 21.3 

g and 1.56g of alpha linolenic acid (omega-3 fatty acid), 

respectively. Cunnane et al., 41 reported that flaxseed supplementation 

increased alpha linolenic acid in the adipose 

tissue without affecting antioxidant status of healthy subjects. 

This report suggests that the antioxidant activity of 

SDG would not have been affected by high alpha linolenic 

acid content in the flaxseed. Dietary alpha linolenic acid 

supplementation reduced fasting plasma glucose (0.20 

mmol/l) in humans. 42 Effects of alpha linolenic acids on 

blood glucose and TG concentrations were not determined 

in the current study. A cumulative effect of alpha linolenic 

acid and SDG working together might have lowered fasting 

serum glucose concentrations. However, effects of alpha 

linolenic acid in flaxseed on fasting serum glucose should 

be investigated in the future to clarify the function of lignan 

in lowering blood glucose and TG concentrations. 

Moreover, effects of flaxseed supplementation on the oxidative 

damage or total antioxidant concentration in human 

subjects still remain to be determined. 

CONCLUSIONS 

In this pilot study flaxseed has been shown to delay 

development of diabetes and/or cardiovascular disease by 


lowering fasting serum glucose and triacylglycerol in glucose 

intolerant individuals. However, further study is necessary 

to test the antioxidant activity of flaxseed using SDG 

in people with impaired glucose tolerance. 

ACKNOWLEDGEMENTS 

This study was supported by grants from North Dakota 

State University Research Development Support Program 

and North Dakota Oilseed Council. The authors thank Drs. 

James Hammond, Dennis Wiesenborn, Frank Manthey, and 

Brady Vick at North Dakota State University for proximate 

or lignan analysis of supplements, and Curt Doetkott and 

Koji Fujiwara for aiding in statistical analyses. The authors 

express special thanks to Dr. Jack Carter, emeritus professor 

at North Dakota State University, for providing his 

expertise in flaxseed, acquiring flaxseed and durum wheat 

bran, arranging proximate and lignan analyses for the study, 

and editing the manuscript. The authors also acknowledge 

North Dakota Oilseed (flaxseed) Council, Bismark, ND, for 

provision of ground full-fat yellow omega flaxseed and 

Dakota Growers Pasta Co., Carrington, ND, for provision of 

durum wheat bran throughout the study. 

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JANA Vol 9 #1 - American Nutraceutical Association

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