“Let food be thy medicine and medicine be thy food.” -Hippocrates
“Clearly, dietetics professionals can no longer evaluate foods solely in terms of macro-and micronutrient content. Consideration of other physiologically active components …will be necessary… In the future, a wide range of select foods may be “prescribed” to enhance the health of an individual. This is a shift from our earlier employed nutrition education approach that focused on limiting intake of foods high in “unhealthful” components such as fat and cholesterol.”
American Dietetic Association, www.eatright.org
“Phytonutrients (.’to-noo’-tree nts) are health promoting compounds found in plants. Zoonutrients (zoh-uh-noo’-tree-uh nts) are health promoting compounds found in animals… that provide health bene.ts beyond the provision of essential nutrients and energy. Functional foods are those that contain one or more of such substances…thought to in.uence health.”
-Diet and Health Trends Concepts and Controversies, University of Idaho http://www.avs.uidaho.edu/avs305/Intro%20to%20nutrients.ppt
“Understanding how molecular structures have evolved to provide nutritional functions in animals will lead to a new generation of foods that can deliver on the promise of maintaining optimal health.”
-Robert E. Ward and J. Bruce, Zoonutrients and Health, Food Technology, Vol 57, March 03, pp 30- 36
“An ounce of prevention is worth a pound of cure.”-Benjamin Franklin, Poor Richard’s Almanac
Functional Foods from Plants and
We have long known that foods contain the nutrients we need to sustain life. Nonethe
less, nutrition and food scientists are continually .nding new bene.cial components in food that help keep us healthy and prevent the development of disease states. Indeed, the very concept of what bene.ts foods can provide is continuously evolving. The previous emphasis on health maintenance through recommended nutrient allowances and dietary guidelines has recently evolved into a focus on the promising use of foods to promote optimal health and reduce the risk of chronic diseases.1 Today, nutritionists describe foods that are rich in certain “ingredients” that may provide a health bene.t beyond the
traditional nutrients it contains as functional foods.These functional food ingredients are sometimes described as “quasi-nutrients” and are best exempli.ed by two main groups, the phytonutrients and the zoonutrients.
A signi.cant direction in the effort to understand the health bene.ts of plant foods characterization of their physiologically active constituents called phytonutrients. ni.cant levels of low molecular weight, secondary metabolites with important role in optimizing human health been appreciated.4 For example, phytonutrients have been demonstrated to provide the following physiological effects:
facilitate cell-to-cell communication, 5
modify cellular receptor uptake of hormones, 6
convert to vitamin A, 7
repair DNA damage from toxic exposure, 8
detoxify carcinogens through the activation of the cytochrome P450 and Phase II liver enzyme systems, 9
serve as antioxidants to help prevent various forms of cancer, 10
cause apoptosis (cell death) in cancer cells, 11
enhance immune response, 12
help prevent cardiovascular disease, 13
help prevent osteoporosis, 14
help prevent macular degeneration and cataracts. 15
Just as plant foods may provide phytonutrients, animal foods may also contain bene.cial substances called zoonutrients. Zoonutrients are food molecules that have
been shown to have potential in modifying multiple physiological functions including anti-in.ammatory, anti-hypertension and antimicrobial actions, stimulation of bene.cial bacteria, the maturation of intestinal cells, and the education of the immune system.16
The currently more familiar anti-in.ammatory, anti-clotting and triglyceride lower ing effects of the omega-3 oils of cold water .sh are largely an attribution of their zoonutrients EPA and DHA.
In Chicago Oct., 1997, the International Whey Conference
reported the following seven .ndings concerning the zoonutrients in whey:
Very low molecular-weight whey peptides may help prevent cancer and heart disease.
Certain whey peptides boost immune status by increasing the body’s main cellular protector, glutathione.
Whey protein, by increasing thymus development, has reduced colon cancer in rats.
One powdered whey supplement in conjunction with photodynamic therapy reduced tumor size by 60% in rats.
Whey contains growth factors (IGF-I and II) proven to assist in gut and wound healing, inside and out.
he protein and peptide zoonutrients in whey and colostrum, and then examine iry based, zoonutrient rich, functional food formula.
have been available for hundreds of years, though mostly they were considered has only been during the last 25 years that new processing methods and scienti.c ins.
otein is often described as a “nutritionally perfect protein” in the sense that it contains and non-essential amino acids required by the human body. Whey’s amino acid y related to the optimal physiological needs of the human body, including an abuncontaining amino acids, all in a highly bio-available form. Whey protein’s quality cribed by such terms as high Biological Value (BV), high Protein Ef.ciency Rat-
high Net Protein Utilization (NPU). Human breast milk is 80% whey protein!
ns are so easily digested and well utilized they may be an ideal source of protein
mino acids for those ill and elderly with declining appetites, challenges with food s then optimal dentition and/or digestive strength. Being low in carbohydrates and whey protein is well suited for restricted carbohydrate diet strategies.
ey protein may be the best candidate for maximizing muscle growth. Whey protein mal balance of amino acids for muscle growth, especially glutamine or glutamic
e. L-Glutamine is the most abundant non-essential free amino acid in the body. on of free glutamine appears to in.uence whether muscle will break down due to ontent or build up via high glutamine content. Athletes suffering from overtraineem to have decreased blood concentrations of glutamine. This may lead in turn of immune functions and slow recovery time following exercise. Even patients
trauma (surgery, burns, stress, chemotherapy, radiation etc.) have an increased
amine. Such increased demand is probably the result of increased use of
he immune, antioxidant, and detoxi.cation or irradiated tissues.
st percentages of branched-chain amino acid
onstrate less muscle loss when the body muscle is being broken down as it is in severe infections,
ng effect occurs because BCAA serve as a d
ds have quickly become the protein of choice
Undenatured whey protein is rich in cystine, the thermo-labile amino acid which represents an ef
cysteine delivery system for the cellular synthesis of glutathione. Both cysteine and glutamine, along with
glycine, are necessary the synthesis of the tri-peptide glutathione (GSH), one of the major detoxi.
(Phase II sulfonation) and antioxidants of the body. Enhancing glutathione levels also helps reduce the risk
of infections by improving white blood cell functions. However, the unique disul.de cystine bon
are heat sensitive (thermo-labile) so only carefully processed, undenatured
whey proteins deliver b
cystine di-peptides for intracellular conversion to cysteine, thus maximizing glutathione levels
important immune, antioxidant, and detoxi.cation bene.ts.17
New preliminary research suggests whey protein may be able to reduce stress and depression b
ing cortisol and increasing brain serotonin, improve liver function in hepatitis, help chronic fat
prove athletic performance and reduce blood pressure.18 Other reports show whey proteins hav
been shown to have positive impact potential in appetite suppression, cholesterol reduction, an
inhibition of dental plaque and dental caries.19 Researchers at the Arkansas Children’s Nutritio
funded by the USDA, have found that whey and soy protein may help prevent breast cancer.20
The Sub-Fractions of Whey Protein
The proteins in whey come in many protein sub-fractions, namely Beta-lactoglobulin, Alpha-lactalbumin, Immunoglobulins(Ig), Bovine Serum Albumin (BSA), Glycomacropeptide (GMP), Lactoferrin, Lactoperoxidase, and Lysozyme. Their relative amounts will vary somewhat, especially depending on how the whey proteins are processed.
Beta-lactoglobulin is the most abundant whey protein component, making up approximately 50-75% of the whey protein. It binds fat-soluble vitamins making them more available to the body. It is rich in muscle sparing energy supplying branched chain amino acids (BCAAs).
Alpha-lactalbumin is the second most abundant whey protein component, making up approximately 12-24% of the whey protein. It is primary protein found in human breast milk. Being high in tryptophan, an essential amino acid, potential bene.ts include sleep regulation and mood improvement under stress. Alpha-lactalbumin is the only whey protein component capable of binding calcium.
Immunoglobulins (mostly IgG, with IgA and IgM), and Bovine Serum Albumin (BSA), make up approximately 5-15% of the whey protein. Immunoglobulins and BSA, the latter a particularly cystine rich sub fraction, are the predominant whey protein components found in colostrum. Immunoglobulins convey passive immunity, especially enterically.
Lactoferrin, a glyco-protein, makes up approximately .2 -1% of the whey protein. Lactoferrin inhibits the growth of bacteria and fungi due to its ability to bind iron, a function known as ferro-privation.21 Iron is a nutrient usually required for bacterial growth. Lactoferrin also promotes the growth of bene.cial bacteria such as L. Bi.dus, helping infants establish good microbial conditions in their intestines, described as “eubiosis”. It is also an antioxidant that naturally occurs in many body secretions such as tears, blood, breast milk, saliva and mucus.22 Lactoferrin has anti-viral, anti-tumor activity, anti-in.ammatory / anti-oxidant activity, and immuno-modulat-23, 24, 25, 26, 27 Lactoferrin is also a cystine rich sub fraction.
Lactoperoxidase makes up approximately 0.5% of the whey protein. Like lactoferrin, it inhibits the growth of iron dependent bacteria.
Lysozyme makes up less than 0.1% of the whey protein. Lysozyme contains immunity enhancing properties.
Glycomacropeptide (GMP) helps control appetite and inhibit the formation of dental plaque and dental cavities. Levels may range from 1% to 18%, depending on how the whey is processed.
Weighing the Different Forms of Whey
Whey Protein Concentrates (WPC) may supply up to 80% protein and usually include medium to comparatively high lactose levels.
Whey Protein Isolates (WPI) and Whey Protein Hydrosylates (WPH) contain little to no appreciable lactose or carbohydrates, are
less that 1% fat, and are usually 90% or more protein!
The advantage of whey protein isolate (WPI) is that it is highest in protein and lowest in lactose. However, some immune peptides,
GMP, vitamins and minerals are lost in an isolate. The advantage of a hydrosolate (WPH) is that it is highest in BCAA, being ap
hydrolysis, making it even easier to digest. But the hydrolysis process also tends to denature the otherwise bioactive peptides in whey.
Whey protein concentrate (WPC) is a more nutritionally whole and complete product, but per gram supplies less protein and more fat,
cholesterol and lactose. Many people are intolerant of the latter. One solution for a dairy based functional food is to combine the low
lactose, low fat, high protein of WPI with the concentrated bioactive peptides of whole colostrum.
As mentioned earlier, to have maximum ef.cacy as a functional food, it is most important that whey peptides are not denatured during any of the above processing.28 We will examine “denatur ization” more fully shortly. Furthermore, some health professionals prefer whey products from cows not given growth hormones, routine prophylactic antibiotics, or GMO or pesticide containing feeds.
The Whey to Weight Loss
Our present understanding of the physiology of appetite is still incomplete. However, we are aware that there are various endogenous brain messenger substances (neuro-transmitters, neuro-hor mones) that effect, among other things, mood, memory, relaxation, focus, and appetite. A better appreciation of the relationship of these brain messenger chemicals to their nutritional building blocks, particularly the amino acids, may well be an important factor in endeavors to modulate appetite and thereby facilitate clinical efforts to attain and maintain optimal lean body mass.
According to Julia Ross, M.D. in her book “The Diet Cure”, published in 2000, one such brain messenger is serotonin, which is known to affect mood, promote a relaxed feeling of “lightness”, support deep sleep and melatonin production, and affect appetite cravings, especially for carbohydrates.
The amino acid “building block” to serotonin is tryptophan. It is therefore important to appreciate that the ratio of tryptophan to large neutral amino acids and branched chain amino acids determines tryptophan’s availability to the brain. The large neutral amino acids (LNNA) are tyrosine and phenyalanine, and the branched chain amino acids (BCAA) are leucine, isoleucine, and valine, all of which compete with tryptophan to crossing the protective blood brain barrier to enter the brain.29
Furthermore, it is important to be aware that insulin, which is released as the result of ingesting carbohydrates, as any serious body builder knows, is anabolic, meaning growth promoting. Insulin facilitates the removal of proteins, speci.cally the aromatic and branched chain amino acids, from the bloodstream to be largely directed to muscle tissue. This function of insulin helps remove these competitors to tryptophan’s crossing the blood brain barrier. Insulin resistance would interfere with this function.
In a 2003 paper reported in the American Journal of Clinical Nutrition30, both the absolute plasma tryptophan concentration and
and the ratio of tryptophan to large neutral amino acids were low. This was true both during
during and after
after the successful weight loss programs, even though said programs were successful in maintaining lean body mass. The authors also noted that the obese participants are oftege neutral amino acids to tryptophan levels. Absolute plasma concentrations of tryptophan as such were also reported to be low in dieting patients before successful weight loss. According to the authors, Breum et al., these two observations, and their potential concomitant effects on mood and appetite, may well be part of the reasons for relapse after diet induced weight loss!
These data strongly support the notion that obese persons may do well to be supplementing tryptophan. As both low levels of tryptophan and its availability to the brain vis-à-vis tryptophan/LNAA ratios tend to lead to below optimal serotonin levels, which is subsequently conducive to carbohydrate cravings, poor sleep, low self esteem and mood, and impulsive behavior, such a notion is not without logical argument.
Unfortunately, the FDA continues to argue what many feel is its unjusti.ed prohibition of tryptophan as a dietary supplement. An alternative, and perhaps even superior, approach may be to utilize high quality whey protein isolates. WPI has both an unusually rich supply of tryptophan and a higher ratio of tryptophan to BCAAs as compared to other proteins such as soy protein or casein. The sifect on tryptophan uptake by the brain and the brain’s neurophysiology.
As WPI is 90% or more highly biologically available and complete protein, with minimal additional calories from carbohydrates and fats, igies. and and magnesium. Additionally, it may be best to not slow down the assimilation of the amino acids with simultaneous ingestion of large amounts of .ber or fats, if our desire is to generate a more immediate effect on mood, sleep, behavior, and appetite.
Health Enhancing Potential of Undenatured Whey Protein Zoonutrients
As we have pointed out previously, whey protein is not only a great source of high quality protein macro-nutrition, rich in complete, balanced, easily absorbed amino acids, it is also a rich source of a multitude of unique zoonutrients (zoh-uh-noo’-tree-uh nts).
As phytonutrients, like lycopene, lutein, phytoestrogens and polyphenols, are non-nutritive phyto-chemicals in plant foods that nonetheless have salubrious metabolic effects in the animals and humans who consume them, even so zoonutrients are similarly ef.cacious ansume them. In 2003, Ward and Bruce write in Food Technology
echnology, “…different peptides from milk have been well described to modify blood pressure, neurologic activity, immune functions, food intake, intestinal functions, and even dental calci.cation… These discoveries have prompted scientists to pursue…how speci.c molecules in foods affect health, how foods modulate the immune system and the interaction between bene.cial bacteria, pathogenic bacteria, and our innate and acquired immune-protective mechanisms.”31
JB German of UC Davis Dept. of Food Science and Technology writes,
“Metabolic products of the lactating mammary gland (milk) are being shown to provide a spectrum of bene.ts… Research to explore the majority of non-essential nutrients was accelerated when investigators began to explore the principles of immunology and physiology in molecular detail using isolated cells, and to assemble screening assays… Surprisingly, components from milk were discovered to exert signi.cant effects on many of these assay systems. This led to the second generation of zoo -nutrients, molecules that modi.ed physiological targets of known health problems including anti-in-.ammatory, anti-hypertension and antimicrobial actions. Now modern biological tools are… deducing dietary functions that were previously unknown, including as examples, the stimulation of bene.cial bacteria, the maturation of intestinal cells and the education of the immune system…(that) will pr the next generation of health properties.”32
However, it is very important to note that the various zoonutrients in milk are more or less “thermo-sensitive” meaning that they are easily deformed or “denatured” by heat, as the following quotes will demonstrate.
“Recent observations have revealed to us that the described biological activity of the whey pr unrelated to its nutritional quality (i.e., related to the zoonutrients), is actually dependent on the undenatured conformation of the proteins.…The immuno-enhancing and the other speci.c biological properties of dietary whey protein…are heat labile and dependant upon the undenatured (native) state of the protein… and are independent of its nutritional quality which is unaltered by the process of denaturation…Our data indicate that the humoral immune response is highest in…a dietary whey protein concentrate exhibiting the highest solubility (undenatured conformation)…this type of whey protein concentrate exhibit(s) higher levels of tissue glutathione (the major endogenous antioxidant and major detoxicant in humans). The presence in the serum albumin fraction of glutamyl-cysteine groups (rare in food protein) and the speci.c intramolecular bond as related to the undenatured conformation of the molecule are considered to be key factors in the glutathione-promoting activity of the (whey) protein mixture.”33
“Milk whey protein fed to mice as a constituent of the diet elicited a systemic humoral response…. The serum antibodies were of the IgG class (Immunoglobulin G)… However, when heat-denatured whey protein was fed, the animals showed only a poor serum re-sponse…”34
“Comparison of IgG content in raw milks and corresponding HTST (High Temperature Short Time)-pasteurized milks of varying fat content indicated 59–76% retention after pasteurization, (or 24% to 31 % loss!- editor)…This study demonstrates the dependence of bovine IgG stability in milk products on severity of thermal treatment used in various commercial processes.”35
“Thedesign of heat treatments of milk in order to preserve the biological function of Ig.”36
“The ef.cacy of …oral administration of Ig from colostrum…provides effective protection against…(bacterial) infections of human otavirus (that) prevents the occurrence of diarrhea and reduces the duration of agent eserve the immunological function of Ig, the effects of processing and storage conditions on stability must be
known. … the ability of (heated) IgG to bind the antigen and, thus, to maintain its immunological activity probably has been overestimated until now. In conclusion, these considerations should be taken into account in the design of heat treatment of milk in order to preserve the biological function of Ig…”37
In conclusion, it is most important when considering “which whey to go” that if one wants to maximize the potential bene.ts of the zoonutrients in a whey protein formula, that one chooses a product that is processed with minimal heat exposure and can demonstrate that all the whey products in the blend are highly undenatured.
Colostrum: Nature’s Most Nutrient Dense Zoonutrient
“many hormones, growth factors and bioactive substances present in the maternal organism are present in colostrum…, often exceeding concentrations that occur in maternal
-Endocrinology Review, 1993
“…colostrum has been used to successfully treat: Thrombocytopenia, Anemia, Neutropenia, Myasthenia Gravis, Guillain Barre Syndrome, Multiple Sclerosis, Systemic Lupus, Rheumatoid Arthritis, Bulluos Pamphigoid, Kawasaki’s Syndrome, Chronic Fatigue
Syndrome and Crohn’s disease, among others.”
-Dr. Dwyer; NEJM
. Although whey protein can be rich in zoonutrients when properly processed, colostr
nature’s most nutrient dense zoonutrient. Robert Preston, MD, President of the Intern Institute of Nutritional Research, de.nes colostrum in the following manner: “When mal)… gives birth to its offspring, its mammary glands .lter out of the blood the imm factors it has acquired through a lifetime of .ghting disease-causing organisms. It th concentrates these factors into special non-milk immune supporting .uid called colo A mother animal produces true colostrum for only the .rst twenty-four hours after g birth.”40
Indeed, besides being very rich in highly bioavailable vitamins and minerals, the col of mammals produced has two main functions: to supply both passive and active imm factors for the otherwise highly susceptible new born, and provide growth factors, n the immune system via the thymus, but for cells throughout the body. Colostrum’s Immune Factors
Immunoglobulins: Immunoglobulins are protein molecules that provide passive imm fects that can be ef.cacious, both prophylactically and therapeutically, against allerg teria (including H. pylori), viruses, parasites, fungi and yeast. The immunoglobulins in colostrum are predominantly IgA with trace amounts of IgD, IgE, IgG, and IgM. H colostrum typically contains 2% IgG content, while whole bovine colostrum can hav techniques can yield as high as 40% colostrum, though such would no longer be a “w and factors are thereby diminished. Immunoglobin concentrations should be veri.ed to verify label claim.
Lactalbumin: These protein molecules are rich in double-bonded cystine which promotes the production of glutathione, the major intracellular endogenous antioxidant and detoxicant. Lactalbumin may also help raise serotonin in de.ciency states and lower cortisol when in excess.
Lactoferrin: Discussed earlier, lactoferrin is most familiar as an iron-binding protein. Lactoferrin’s competition for available iron in the gut both inhibits bacterial and viral populations and oxidation, and the resulting “down stream” in.ammation, from excess iron radicals.
Lysozymes: Lysozymes contain enzymes that can attach to and digest bacteria cell walls, thus destroying them.
Glycoproteins: These sugar-amino complexes act as protease and trypsin inhibitors, thus protecting the immune and growth factors in colostrum which are otherwise vulnerable to degradation via enzymatic action.
Proline-rich Polypeptides (PRP): These are small, very low weight molecules (6,000 Daltons) that have an active immune modulating effect upon the thymus. They have been variously described as “biological response modulators, “info-peptides”, “transfer factors” and “colostrinin”.41, 42 Immunode.ciency (Th1 de.cit states), as in HIV, EBV and herpes, may be thus counteracted, while immune hyperactivity (Th2 hyperactivity), as in autoimmune and allergic diathesis conditions, may be inhibited.43 PRPs isolated from colostrum and taken sublingually have shown great promise as an immune equilibrating nutraceutical and as a potential therapy for Alzheimer’s disease! 44
Thtween 1-3% of the total powder weight. However, most manufacturers of colostrum powders remove much of the PRP fraction, along with lactose, minerals and water, using ultra-.ltration technology, to elevate the passive immunity supporting immune globulins and the protein content of the powder. This then reduces the active immunity modulation effectiveness of the colostrum powder. Patented technologies now exist that can fortify whole colostrum such that it provides 6% PRPs by weight!
Cytokines: Cytokines are integral to intercellular communications that regulate immune activity and related down stream in.ammatory responses. These immune messengers include the interleukins, the lymphokines, and interferon. The PRP’s mentioned above have a modulating effect on the cytokines.
Colostrum’s Growth Factors
Growth factors from mammalian bovine colostrum are by and large not species speci.c. Indeed, they are almost identical to human colostrum! The various growth factors in whole colostrum are by de.nition anabolic, stimulating both generation and regeneration of epithelial, mesenchymal, and endothelial
endothelial cells. During periods of low calorie intake, growth factors favor the use of fat for fuel and are therefore protein and “lean body mass” sparing.
Insulin-like Growth Factor I and II: Often abbreviated as IGF-I / IGF-II, these
are the predominant growth factors in colostrum. They help regulate lipid, protein and carbohydrate anabolism. Of note, IGF-I is one of a handful of molecules that promotes the growth and repair of DNA and RNA. Epithelial Growth Factor: EGF enhances dermal anabolism. Indeed, topical applications of EGF concentrates may soon be a common cosmeceuticals ingredient. Colostrum as such already is.
Transforming Growth Factors A & B: TGF A & B promote mesenchymal cellular proliferation. TGF thus has potential for assisting bone and cartilage repair, deep wound healing, and restoring intestinal integrity in “leaky gut” syndromes. Platelet-Derived Growth Factor: PDGF promotes growth not only in connective tissue such as .broblasts and smooth muscle, but has some promise in sparing and regenerating nerve tissue as well.
As with most nutraceuticals, quality, and therefore clinical ef.cacy, greatly varies. According to Andrew Keech, PhD, of Advanced Protein Systems
, some things to look for when choosing a colostrum product include:
HPLC Analysis: Make sure there is High Performance Liquid Chromatography (HPLC) analysis on every batch to verify label claims for IgG percent.
Low Heat Processing: Avoid high heat pasteurization, .ash pasteurization without immediate cooling to 40 degrees F, and direct drying processes which all denature colostrum’s peptides.
First and Second Milkings: The .rst and second milkings, taken within the .rst 24 hours of calving, are the richest in IgG and protein peptides. The best products use only these early milkings.
Antibiotic/Hormone “Free”: The most desirable colostrum is from cows not routinely treated with prophylactic antibiotics or given synthetic growth hormones.
Solubility: Clients and patients will prefer a colostrum powder that dissolves quickly without clumping.
Freshness: When possible, prefer colostrum for cows whose climate conditions allow year-round production. Know that most colostrum is produced only once a year because calving typically occurs in the spring. As always, prefer manufacturers that follow “Good Manufacturing Practices” (GMPs). This should include a microbiological analysis on each product batch.
dosing may be too “energizing” for some persons. Colostrum should not be taken with protein digesting enzymes which may denature the peptides.
The Physiological Functions of Proline Rich Polypeptides (PRP)
Proline Rich Polypeptides are perhaps the most powerful of colostral peptides vis-à-vis active immune modulation and therefore desenonetheless have a very powerful effect in initiating and balancing our immune responses.
Proline Rich Polypeptides, also known as PRP, enhance the ability of the thymus gland to release factors that help regulate immune functions in the body. Speci.cally, certain T cells, called Th1 helper cells, are antagonist to the activity of Th2 helper cells that promote certain functions of B lymphocytes. PRP can induce a shift from a predominantly humeral
humeral immune response to a more protective cellular response described as a “Th2 to Th1 shift”. Doing so may assist the immune system in more effectively .ghting chronic viral and bacterial infections while simultaneously inhibiting the initiation of inappropriate in.ammatory cascades associated with allergy, chemical sensitivity and auto-immune responses.
A more detailed review of some of the main physiological functions of proline rich polypeptides follows.
Modulate the immune system
– PRP promote T-lymphocyte function and can either stimulate the lymphocytes to become helper T-cells or suppressor T-cells.45, 46, 47 Helper T-cells activate B-lymphocytes by presenting an antigen (such as a viral protein) to the B-cell, which then produces antibodies to that protein.48 Helper T-cells also help produce memory T-cells which retain the “memory” of the antigen to shorten the response time in case of new infection.49 Suppressor T-cells deactivate other lymphocytes, effectively turning off the immune response to avoid damage to healthy tissue.50 PRP also stimulate the production of a whole range of cytokines, particularly the pro-in.ammatory cytokines TNF-alpha and INF- gamma and the anti-in.ammatory cytokines IL-6 and IL-10 .51, 52
Act as molecular signaling devices
– PRP work through speci.c receptors on cell surfaces.53
Stimulate undifferentiated lymphocytes in thymus to become either helper T-cells or suppressor T-cells
– PRP from ovine (sheep) colostrum act as a hormone in the thymus gland by stimulating thymocytes (immature lymphocytes) to differentiate and become activated as either helper T-cells or suppressor T-cells.54 Helper T-cells are a vital part of the immune response which stimulate the production and differentiation of cytotoxic T-cells and B-cells, attract white blood cells, and stimulate macrophages to engulf and destroy pathogens. Suppressor T-cells inhibit the production of cytotoxic T-cells to prevent tissue damage and suppress the immune response when no longer needed.
Promote growth and differentiation of B-cells
– PRP promote the growth and differentiation of B-cells, a type of lymphocyte which produces antibodies to antigens, including viral antigens.55
Stimulate Natural Killer cell (NK cell) activity
– PRP stimulate the activity of NK cells up to 10 times, far greater than any other known substance. NK cells, along with cytotoxic T-cells, are the cells which actually attack and kill pathogens. NK cells also attack and kill cancerous cells. 56
Stimulate the production of tumor necrosis factor-alpha (TNF-a) and interferon-gamma (INF-y)
– PRP stimulate production of pro-in-.ammatory cytokines TNF-a and INF-y, the two major pro-in.ammatory cytokines, in white blood cells, peritoneal cells, and placental and amniotic membranes.57, 58, 59
Promote the proliferation of leukocytes
(white blood cells)60
Stimulate production of cytokines by peripheral blood cells
-The types of cytokines stimulated by PRP depend on the antigenic stimulation present or the activity state of the immune system (under-productive or over-productive). In one study, mice exposed to herpes simplex virus (HSV) were stimulated to produce large amounts of IL-2 and INF-y and small amounts of IL-10, while mice which had been given transfer factor (PRP) prior to infection responded to HSV by secreting INF-y but no IL-2. PRP stimulates the production of TNF-a, INF-y, IL-6 and IL-10 in blood cell cultures.61, 62
Induce differentiation and maturation of monocytes and macrophages
Increase the permeability of blood vessels in the skin
– Part of the in.ammatory response to infection is an increase in the permeability of blood vessels in the skin to allow the passage of blood cells and cytokines into the connective tissue to combat the infection. PRP is known to initiate this in.ammatory response.64
Produce immunity to certain viruses
– PRP has been experimentally shown to provide immunity to several viruses, including herpes viruses , Epstein-Barr virus , HIV , measles , vesicular stomatitis virus (a close relative of the rabies virus), and others.65, 66, 67, 68, 69, 70, 71, 72, 73
Inhibit viruses known to be associated with autoimmune diseases Epstein-Barr virus and human herpes virus-6 (HHV-6) has been associated with chronic fatigue syndrome, an autoimmune disorder PRP inhibits the replication of both viruses.74, 75
May help down regulate the “cytokine storm” seen in bird .
.uenza A virus subtype H5N1 sets off a so-called “cytokine storm” which usually results in an often fatal respiratory disease in those infected with the virus. Research has indicated that the storm is caused by cytokine dysregulation which allows pro-in.ammatory cytokines to be produced in large numbers, setting off a potentially fatal in.ammatory response.76 As PRP is known to down regulate an overactive immune system, it potentially could be used to put a stop to the overproduction of cytokines and restore homeostasis to the body, preventing a fatal outcome.
Increase T-cell count in AIDS to normal or near-normal levels- In clinical studies conducted in the nations of Nigeria, Kenya and Zambia in Africa, where AIDS is a particularly devastating disease, PRP oral spray products were shown to boost T-cell (CD4+) levels to normal or near-normal levels in AIDS patients whose T-cell levels prior to treatment were well below normal. Along with the increase in T-cells came a remission of AIDS symptoms within two days of start of treatment, including nausea, vomiting and diarrhea. In the Nigerian study, weight gains of 5% were recorded. Patients taking the PRP spray fared much better in terms of quality of life than did patients on anti-retroviral drugs.77 Thus the ability of PRP to stimulate the immune response when it is insuf.cient by inducing the production of new helper T-cells may enable the immune system of AIDS patients to recover suf.ciently so that it is able to .ght the HIV on its own.
When a mammal is healthy it has an essentially full complement of immune peptides. However, old age, bodily injury, environmental toxins, substance abuse, poor nutrition, excessive stress, suppressed immunity, medication, or illness may result in failure to maintain an optimbe reintroduced into the body. Fortunately, these cells then “instruct” cells to create more copies of the peptides. That is why PRPs are also referred to as “cytokine precursors” or “immuno-modulating peptides.” Thus, reintroduction of a small amount, even perhaps a singcells in the body to their normal amount of the peptide(s) in question.78
Proline Rich Polypeptides are not species speci.c. PRP from bovine milk works on all mammals, including humans, dogs and cats.79 As PRP is produced by all mammals and is an entirely natural product, it is generally thought to be safe for all ages. However, lactose is usually associated with PRP and therefore those with milk intolerance may need to proceed with caution. The addition of lactase, the milk sugar digesting enzyme, may ameliorate lactose intolerance.
Also, delicate immune system changes occur following conception and during pregnancy. Speci.cally, there is a shift to Th2 dominance to inhibit the mother’s immune system from over responding to the different DNA of the new life now inside her. Although there are no known reports of colostrum’s interference with full and normal gestation, until further investigation assures safety, caution suggests pregnant women and women hoping to conceive should avoid PRP rich colostrum products unless suggested by their doctor.
Whey Protein Synergists: Glutathione, Selenium and Inulin
Nutrients and other bioactive food components that occur naturally in foods act synergistically with other dietary elements…dietetics professionals have a unique opportunity to promote whole foods…. In addition…functional food products can be developed that further enhance the health bene.ts of food…a combined functional food and food supplement approach may afford the greatest protection….”
-J Am Diet Assoc. 1999; 99(10):1278-1285 Reduced Glutathione
As stated earlier, undenatured whey protein’s immunological effect is in no small part likely related to the glutamylcysteine groups which act as the substrate for glutathione (GSH) synthesis. These cystine groups needed for the intracellular conversion to cysteine are in whey and colostral sub-fractions.80 However, this bioavailable, double bonded cystine portion is very thermo-labile. Denaturization by heat will therefore greatly inhibit the ability of whey proteins to act as precursors to GSH synthesis, though not affecting the biological value of whey as a protein nutrient as such.81
Glutathione itself is a non-essential nutrient composed of three amino acids: glutamic acid, glycine and cysteine, or more exactly the tripeptide L-gamma-glutamyl-L-cysteinylglycine. Availability of cysteine is a limiting factor in the liver’s synthesis of glutathione.82 Monomeric glutathione is also known as reduced glutathione and its dimer is also known as oxidized glutathione AKA glutathione disul.de. 90% of the glutathione in healthy living cells is in the reduced form.83
Glutathione is widely found in all forms of life and plays an essential role in the health of organisms, particularly aerobic ones. In humans, animals, and plants, glutathione is the predominant non-protein thiol
thiol and functions as an antioxidant, keeping its own -SH groups and related proteins in a reduced condition.84
Glutathione is a cofactor for glutathione S-transferases, enzymes which are involved in the detoxi.cation of xenobiotics, including carcinogens. It is also a cofactor for the glutathione peroxidases, which are crucial selenium-containing antioxidant enzymes. It is also involved in the regeneration of ascorbate from its oxidized form, dehydroascorbate. There are undoubtedly multiple functions for glutathione yet to be appreciated.85
Glutathione is present in the diet in amounts usually less than 100 milligrams daily. It does not appear that much of the oral intake is absorbed from the intestine into the blood, at least in humans. However, there is an occasional study that does show an increase in circulating glutathione after oral administration.86, 87, 88, 89 There is greater evidence that glutathione may be absorbed into the enterocytes where it may help repair damaged cells.90 Patents have been submitted for reduced glutathione in a liposome claiming enhanced absorption.91
Chronic functional glutathione de.ciency is associated with immune disorders, an increased incidence of malignancies, and in the case of HIV disease, probably accelerated pathogenesis of the disease.92, 93 Acute manifestations of functional glutathione de.ciency can be seen in those who have taken an over-dosage of acetaminophen (TylenolTM). This results in depletion of glutathione in the hepatocytes, leading to liver failure and death, if not promptly treated.94
Supplemental doses ranges from 50 to 200mg. Oral doses of up to 600 milligrams daily are well tolerated. There are no reports of adverse reactions or over-dosage.95
Glutathione formation, which we have shown is synergistic with some of whey protein’s potential bene.ts, requires an adequate level of selenium. Mammals de.cient in selenium have markedly decreased glutathione dependent peroxidase activity. Some milk, especiaintake, the recommended levels of selenomethionine just to support the WPI intake is 8 to 12 mcg.96 However, more optimal levels of selenium are also thought to confer bene.ts in its own right.
Selehuman avitamin C from its oxidized state.97
Selenium is found in human and animal tissues as L-selenomethionine or Lselenocysteine. As mentioned, these selenoproteins are essential to the endogenous production of the antioxidant proteins, speci.cally the four glutathione peroxidases (GSHPx 1-4). In addition to its antioxidant activity, selenium may also have immuno-modulatory, anti-carcinogenic and anti-atherogenic activi98, 99, 100 It may have activity in detoxi.cation of some metals and other xenobiotics, as well as activity in fertility enhancement in males.101, 102
Selenium de.ciency appears to depress the effectiveness of various components of the immune system. Selenium supplementation in humans has resulted in increased natural killer cell activity.103 Still, the immuno-modulatory effects of selenium are not well understood.
The povitro, selenium has been shown to up-regulate apoptosis in tumor cells and increase macrophage killing and protect against oxidative DNA damage.104
There appears to be an inverse relationship between coronary heart disease and selenium intake. The possible anti-atherogenic activity of selenium may be accounted for, in part, by its antioxidant activity. Glutathione peroxidase may protect low density lipoprotein (LDL) from oxidation, thereby inhibiting atherogenesis and platlet aggregation. (
(LDL) from oxidation, thereby inhibiting atherogenesis and platlet aggregation. (Lipoperoxides impair prostacyclin synthesis and promote thromboxane synthesis).105
Selenium has been demonstrated to antagonize the effects of a number of toxic metals, including cadmium and arsenic.106
L-selenomethionine is ef.ciently absorbed from the small intestine via a similar mechanism to that of L-methionine. L-selenomethio-nine is transported via the portal circulation to the liver where a fraction is extracted by the hepatocytes and the remaining amount is transported by the circulation to the various tissues of the body.
Intakesof doses of 1,000 micrograms (or one milligram) or greater daily may cause adverse reactions. The most frequently reported adverse reactions of selenosis (chronic selenium toxicity) are hair and nail brittleness and loss. Other symptoms include skin rash, garlic-like breath odor, fatigue, irritability and nausea and vomiting.107
Selenium is abundant in garlic, onion, broccoli, whole grains and most especially Brazil nuts. The average daily intake of selenium in the United States is reported to be from 60 to 100 micrograms. Optimal daily dosage may be 200 mcg a day.108
Inulins are naturally occurring fructose-containing oligosaccharides (FOS) which are of a class of carbohydrates known as fructans. Inulins aasparagus, chicory and artichokes. The latter two are the source for most supplemental and functional food sources of inulin. They are sweet tasting and have a smooth texture. As such, inulins are often added to various foods. Inulin intake in the U.S. ranges from 1 to 4 grams daily. It is higher in the European diet.109
Inulins, being largely soluble .ber, are only slightly digested in the small intestine, their energy content being less then half that of digestible carbohydrates. Non-digestible soluble .bers, typically FOS, that promote the growth of bene.cial bacteria in the colon are called prebiotics. The FOS is fermented by a limited number of colonic bacteria, especially bi.dobacteria. Bi.dobacteria may inhibit the growth of pathogenic bacteria, such as Clostridium perfringens and diarrheogenic strains of Escherichia coli. Bi.dobacteria and some other bacteria produce the short-chain fatty acids acetate, propionate and butyrate. Energy, in the form of ATP, is produced from the catabolism of butyrate, an important respiratory fuel for the colonocytes.110, 111
Inulins may have antitumor, antimicrobial, hypolipidemic and hypoglycemic actions.112, 113, 114 They may also help to improve mineral absorption and balance and may therefore have anti-osteoporotic activity.115, 116 These actions compliment many of the potential bene.ts of whey proteins and colostral peptides. Inulin also adds desirable taste and texture bene.ts to functional food drinks.
Doses up to 10 grams daily are well tolerated. Higher doses may cause such gastrointestinal symptoms as .atulence, bloating and diarrhea.
Liposomes, “fat bodies” in Greek, have been known within the scienti.c community for decades. Liposomes are nanosized, self-assembling, membrane-bound mini-capsules composed of phospholipids and related compounds.117 Phospholipids are unique in that they are bipolar molecules with one end being hydrophilic (“water loving”) and the other end being lipophilic (“fat-loving”). This means that the liposomes will carry both hydrophilic and lipophilic molecules equally well. The bipolarity of the phospholipid molecules is what accounts for the self-assembly of the membrane when the phospholipids are introduced into a watery environment.118
Since the mid-1960’s, liposomes have been recognized as effective carriers of biologically active ingredients to target speci.c sites of action in a variety of therapeutic applica
nutrients, peptides, and proteins.124, 125
mize this problem.
of scienti. c evidence to ensure appropriate integration into a varied diet.”
1) American Dietetic Association Position Paper on Functional Foods Abstract, J Am Diet Assoc. 2004; 104:814-826.
2) Diet and Health Trends Concepts and Controversies, University of Idaho http://www.avs.uidaho.edu/avs305/Intro%20to%20nutrients.ppt
3) Hasler C., Blumberg J, Functional Foods for Health Program, Department of Food Science and Human Nutrition, University of Illinois, Urbana, IL and Mayer J, USDA Human Nutrition
Research Center on Aging, Tufts University, Boston, MA
4) Maher J. Scienti.c Phytonutrition and the Standard American Diet: A Proposed Realistic Solution, Part I, Dynamic Chiropractic, June 3, 2002, Volume 20, Issue 12
5) Kelly GS. Larch arabinogalactan: clinical relevance of a novel immune-enhancing polysaccharide. Altern Med Rev 1999; 4(2):96-103.
6) Potter SM. Overview of proposed mechanisms for the hypo-cholesterolemic effect of soy. J Nutr 1995 Mar; 125(3 Suppl):606S-11S.
7) Shils ME, et al. Modern nutrition in health and disease: 8th ed. Philadelphia: Lea & Febiger 1994. p.290.
8) Jenkinson AM, et al. The effect of increased intakes of polyunsaturated fatty acids and vitamin E on DNA damage in human lymphocytes. FASEB 1999 Dec; 13(15):2138-42.
9) Persky V, Van Horn L. Epidemiology of soy and cancer: perspectives and directions. J Nutr 1995; 125(3 Suppl):709S-12S.
10) Steinmetz KA, Potter JD. Vegetables, fruit and cancer prevention: a review. J Am Diet Assoc 1996 Dec; 96(10):1027-39.
11) Mo H, Elson CE. Apoptosis and cell-cycle arrest in human and murine tumor cells are initiated by isoprenoids. J Nutr 1999 Apr; 129(4):804-13.
12) Zhang R, et al. Enhancement of immune function in mice fed with high doses of soy daidzein. Nutr Cancer 1997; 29:24-8.
13) Gaziano JM, et al. A prospective study of consumption of carotenoids in fruits and vegetables and decreased cardiovascular mortality in the elderly. Ann Epidemiol 1995 Jul;5(4):255-60.
14) Head KA. Ipri.avone: an important bone-building iso.avone. Altern Med Rev 1999 Feb;4(1):10-22.
15) Seddon JM, et al. Dietary carotenoids, vitamin A, C, and E, and advanced age-related macular degeneration: eye disease case-control study group. JAMA 1994 Nov 9;272(18):1413-20.
16) J. B. GERMAN, Dept. of Food Science & Technology, Univ. of California, Davis, 1 Shields Ave., Davis, CA 95616
17) Douglas Jr. FW, Greenberg R, Farrell, Jr. HM, and Edmondson LF; Effects of Ultra-High-Temperature Pasteurization on Milk Proteins; J. Agric. Food Chem. 1981, 29, 11-15
18) http://www.usdec.org/.les/Publications/11CARDIOVASCULAR.pdf, http://fst.osu.edu/People/HARPER/Functional-foods/2003%20%20%20BIOLOGICAL%
Life Extension, Jan. 2002, http://www.lef.org/magazine/mag2002/jan2002_report_whey_01.html
19) Whey Protein Institute, http://www.wheyo.ife.org/bene.ts.cfm whey, http://www.ajcn.org/cgi/content/abstract/81/4/792,
20) Cancer, Epidemiology, Biomarkers and Prevention, Journal of the American Association for Cancer Research, January 2000 Vol. 9, 113-117.
23) Arnold, et al. Anti-adenovirus activity of milk proteins: lactoferrin prevents viral infection. Antiviral Res, 2002, 53, 153-8–
24) Tsuda, et al. Cancer prevention by bovine lactoferrin and underlying mechanisms: a Cancer prevention by bovine lactoferrin and underlying mechanisms: a review of experimental and clinical
studies, Biochem Cell review of experimental and clinical studies. Biol, 2002, 80, 131-6, from National Cancer Center Research Institute, Tokyo, Japan
25) Togawa, et al. Lactoferrin reduces colitis in rats via modulation of the immune system and correction of cytokine imbalance., Am J Physiol Gastrointest Liver Physiol, 2002, 283, G 187-95 21
26) Weinberg E D. Administration of exogenous human or bovine Lf to hosts with various infected or in.amed sites has resulted in prophylactic or therapeutic effects. – Human lactoferrin: a novel
therapeutic with broad spectrum potential. J Pharm Pharmacol, 2001, 53, 1303-10
27) Immune Modulation: an overview According to Brock J (1995) Immunology Today 16, 417-419
28) Bounous G, Gold P. The biological activity of undenatured dietary whey proteins: role of glutathione, Clin Invest Med. 1991 Aug;14(4):296-309.
29) Pearson D, Shaw S. Fat and Happy? Tryptophan Concentrations Reduced in Obesity, Life Extension News, Vol. 7 No. 1, Feb 2004
30) Breum et al. Twenty-four-plasma tryptophan concentrations and ratios are below normal in obese subjects and are not normalized by substantial weight reduction. Am J Clin Nutr 77:1112-8 (2003)
31)Ward RE, Bruce J. Zoonutrients and Health, Food Technology, Vol 57, March 03, pp 33
32) J. B. German. History and situation analysis of zoonutrients in nutrition and food industry, Dept. of Food Science & Technology, Univ. of California, Davis, 2001
33) Ibid. 28, pp 296-309
34) Enomoto A, Konishi M, Hachimura S, and Kaminogawa S. Milk Whey Protein Fed as a Constituent of the Diet Induced Both Oral Tolerance and a Systemic Humoral Response, While Heat-
Denatured Whey Protein Induced Only Oral Tolerance , Department of Agricultural Chemistry, The University of Tokyo, Tokyo, Japan , Pub Med, May 2002
35) Li-Chan E, Kummer A, Losso J N, Kitts D D, and Nakai S. Stability of bovine immunoglobulins to thermal treatment and processing , Department of Food Science, The University of British
Columbia, 6650 North West Marine Drive, Vancouver, BC, Canada V6T 1Z4 , 4/94
36) Dominguez E, Perez MD, and Calvo1 M. Effect of Heat Treatment on the Antigen-Binding Activity of Anti-Peroxidase Immunoglobulins in Bovine Colostrum, 1997 J Dairy Sci 80:3182–3187 3182
37) Ibid, E Dominguez, et. Al.
38) Grosvenor CE, Picciano MF, and Baumrucker CR., Hormones and growth factors in milk, Endocrinology Review, 1993 Vol 14, 710-728
39) Dwyer, J.M. Manipulating the Immune System with Immune Globulin, New England Journal of Medicine, April 21, 1994. Vol 330 p1129 (7).
40) 39) Preston, R. Product Review. International Institute of Nutritional Research, pp. 1-4, 1987.
41) Kruzel ML, Janusz M, Lisowski J, Fischleigh RV, Georgiades JA.,Towards an understanding of biological role of colostrinin peptides. J Mol Neurosci. 2001 Dec;17(3):379-89.
42) Kirkpatrick CH. Structural nature and functions of transfer factors. Ann N Y Acad Sci. 1993 Jun 23;685:362-8.
43) Staroscik, et. al. Immunologically active nonapeptide fragment of a proline-rich polypeptide from ovine colostrum: amino acid sequence and immunoregulatory properties. Mol Immunol. 1983
44) Leszek J, Inglot AD, Janusz M, Lisowski J, Krukowska K, Georgiades JA. Colostrinin: a proline-rich polypeptide (PRP) complex isolated from ovine colostrum for treatment of Alzheimer’s
disease. A double-blind, placebo-controlled study. Arch Immunol Ther Exp (Warsz). 1999;47(6):377-85.
45) Zimecki M, Artym J. Therapeutic properties of proteins and peptides from colostrum and milk. Postepy Hig Med Dosw, 59:309-323 (2005).
46) Janusz M, Staroscik K, Zimecki M, Wieczorek Z, Lisowski J. A proline-rich polypeptide (PRP) with immunoregulatory properties isolated from ovine colostrum. Murine thymocytes have on
their surface a receptor speci.c for PRP. Archivum immunologiae et therapiae experimentalis (Warszava) 34(4):427-436 (1986).
47) Wieczorek Z, Zimecki M, Spiegel K, Lisowski J, Janusz M. Differentiation of T cells into helper cells from immature precursors: identi.cation of a target cell for a proline-rich polypeptide
(PRP). Archivum immunologiae et therapiae experimentalis (Warszava) 37(3-4):313-322 (1989).
48) Bishop GA, Haxhinasto SA, Stunz LL, Hostager BS. Antigen-speci.c B-lymphocyte activation. Critical Reviews in Immunology 23(3):159-197 (2003).
49) Shi M, Hao S, Chan T, Xiang J. CD4+ T cells stimulate memory CD8+ T cell expansion via acquired pMHC I complexes and costimulatory molecules, and IL-2 secretion. Journal of
Leukocyte Biology (2006).
50) Zimecki M, Staroscik K, Janusz M, Lisowski J, Wieczorek Z. The inhibitory activity of a proline-rich polypeptide (PRP) on the immune response to polyvinylpyrrolidone (PVP). Archivum
immunologiae et therapiae experimentalis (Warszava) 31(6):895-903 (1983).
51) Inglot A.D, Janusz M, Lisowski J. Colostrinine: a proline-rich polypeptide from ovine colostrum is a modest cytokine inducer in human leukocytes. Archivum immunologiae et therapiae
experimentalis (Warszava) 44(4):215-224 (1996).
52) Zablocka A, Janusz M, Rybka,K, Wirkus-Romanowska I, Kupryszewski G, Lisowski J. Cytokine-inducing activity of a proline-rich polypeptide complex (PRP) from ovine colostrum and its
active nonapeptide fragment analogs. European Cytokine Network 12(3):462-467 (2001).
53) Ibid, 45, pp 309-323
54) Wieczorek Z, Zimecki M, Spiegel K, Lisowski J, Janusz M. Differentiation of T cells into helper cells from immature precursors: identi.cation of a target cell for a proline-rich polypeptide
(PRP). Archivum immunologiae et therapiae experimentalis (Warszava) 37(3-4):313-322 (1989).
55) Julius MH, Janusz M, Lisowski J. A colostral protein that induces the growth and differentiation of resting B lymphocytes. Journal of Immunology 140(5):1366-371 (1988).
56) See DM, Khemka P, Sahl L, Bui T, Tilles JG. The role of natural killer cells in viral infections. Scandinavian Journal of Immunology 46(3):217-224 (1997).
57) Inglot A.D, Janusz M, Lisowski, J. Colostrinine: a proline-rich polypeptide from ovine colostrum is a modest cytokine inducer in human leukocytes. Archivum immunologiae et therapiae
experimentalis (Warszava) 44(4):215-224 (1996).
58) Blach-Olszewska Z, Janusz M. Stimulatory effect of ovine colostrinine (a proline-rich polypeptide) on interferons and tumor necrosis factor production by murine resident peritoneal cells.
Archivum immunologiae et therapiae experimentalis (Warszava) 45(1):43-47 (1997).
59) Domaraczenko B, Janusz M, Orzechowska B, Jarosz W, Blach-Olszewska Z. Effect of proline rich polypeptide from ovine colostrum on virus replication in human placenta and amniotic membrane
at term; possible role of endogenous tumor necrosis factor alpha. Placenta 20(8):695-701 (1999).
60) Kruzel ML, Janusz M, Lisowski J, Fischleigh RV, Georgiades JA. Towards an understanding of biological role of colostrinin peptides. Journal of Molecular Neuroscience 17(3):379-389 (2001).
61) Alvarez-Thull L, Kirkpatrick CH. Pro.les of cytokine production in recipients of transfer factors. Biotherapy 9(1-3):55-59 (1996).
62) Ibid. 52, pp 462-467
63) Kubis A, Marcinkowska E, Janusz M, Lisowski J. Studies on the mechanism of action of a proline-rich polypeptide complex (PRP): effect on the stage of cell differentiation.
Peptides 26(11):2188-2192 (2005).
64) Janusz M, Lisowski J. Proline-rich polypeptide (PRP)–an immunomodulatory peptide from ovine colostrum. Archivum immunologiae et therapiae experimentalis (Warszava) 41(5-6):275-279 (1993).
65) Pizza G, Meduri R, De Vinci C, Scorolli L, Viza D. Transfer factor prevents relapses in herpes keratitis patients: a pilot study. Biotherapy 8(1):63-68 (1994).
66) Pizza G, Viza D, De Vinci C, Palareti A, Cuzzocrea D, Fornarola V, Baricordi, R. Orally administered HSV-speci.c transfer factor (TF) prevents genital or labial herpes relapses. Biotherapy 9(1-3):67-72
67) Meduri R, Campos E, Scorolli L, De Vinci C, Pizza G, Viza D. Ef.cacy of transfer factor in treating patients with recurrent ocular herpes infections. Biotherapy 9(1-3):61-66 (1996).
68) Prasad U, bin Jalaludin MA, Rajadurai P, Pizza G, De Vinci C, Viza D, Levine PH. Transfer factor with anti-EBV activity as an adjuvant therapy for nasopharyngeal carcinoma: a pilot study.
Biotherapy 9(1-3):109-115 (1996).
69) Raise E, Guerra L, Viza D, Pizza G, De Vinci C, Schiattone ML, Rocaccio L, Cicognani M, Gritti F. Preliminary results in HIV-1-infected patients treated with transfer factor (TF) and zidovudine
(ZDV). Biotherapy 9(1-3):49-54 (1996).
70) Ferrer-Argote VE, Romero-Cabello R, Hernandez-Mendoza L, Arista-Viveros A, Rojo-Medina J, Balseca-Olivera F, Fierro M, Gonzalez-Constandse R. Successful treatment of severe complicated
measles with non-speci.c transfer factor. In Vivo 8(4):555-557 (1994).
71) Orzechowska B, Janusz M, Domaraczenko B, Blach-Olszewska Z. Antiviral effect of proline-rich polypeptide in murine resident peritoneal cells. Acta Virologica 42(2):75-78 (1998).
72) van Hooijdonk AC, Kussendrager KD, Steijns JM., In vivo antimicrobial and antiviral activity of components in bovine milk and colostrum involved in non-speci.c defense. British Journal of
Nutrition 84(Suppl 1):S127-34 (2000).
73) Ushijima H, Dairaku M, Honnma H, Mukoyama A, Kitamura T. Immunoglobulin components and anti-viral activities in bovine colostrum. Kansenshogaku Zasshi 64(3):274-279 (1990).
74)Ablashi DV, Levine PH, De Vinci C, Whitman JE Jr, Pizza G, Viza D. Use of anti HHV-6 transfer factor for the treatment of two patients with chronic fatigue syndrome (CFS). Two case reports.
Biotherapy 9(1-3):81-86 (1996).
75) De Vinci C, Levine PH, Pizza G, Fudenberg HH, Orens P, Pearson G, Viza D. Lessons from a pilot study of transfer factor in chronic fatigue syndrome. Biotherapy 9(1-3):87-90 (1996).
76) Chan MC, Cheung CY, Chui WH, Tsao SW, Nicholls JM, Chan YO, Chan RW, Long HT, Poon LL, Guan Y, Peiris JS. Proin.ammatory cytokine responses induced by in.uenza A (H5N1) viruses
in primary human alveolar and bronchial epithelial cells. Respiratory Research 6:135 (2005).
77) Keech, A. Advanced Protein Systems, Unpublished data. (2006).
78) “Novel colostral fractionation process, a method (100) and a system (200) for processing mammalian bodily .uids to isolate target peptides and proteins.” United States Patent 20050092684:
79) Khan, A. Non-speci.city of transfer factor. Annals of Allergy 38(5):320-322 (1977).
80) Beaulieu J, Dupont C, Lemieux P, Whey proteins and peptides: bene.cial effects on immune health, January 2006, Vol. 3, No. 1, Pages 69-78
81) Ibid., 28, pp 296-309
82) Grif.th OW. Biologic and pharmacologic regulation of mammalian glutathione synthesis. Free Rad Biol Med. 1999; 27:922-935.
83) Hwang C, Sinskey AJ, Lodish HF. Oxidized redox state of glutathione in the endoplasmic reticulum. Science. 1992; 257:1496-1502.
84) Sies H. Glutathione and its role in cellular functions. Free Rad Biol Med. 1999; 27:916-921.
85) Ibid., 84, pp 916-921
86) Flagg EW, Coates RJ, Eley JW, et al. Dietary glutathione intake in humans and the relationship between intake and plasma total glutathionine level. Nutr Canc. 1994; 21:33-46.
87) Witschi A, Reddy S, Stofer B, Lauterburg BH. The systemic availability of oral glutathione. Eur J Clin Pharmacol. 1992; 43:667-669.
88) Hagen TM, Wierzbicka GT, Sillau AH, et al. Bioavailability of dietary glutathione: effect on plasma concentration. Am J Physiol. 1990; 259(4 Pt 1):G524-G529.
89) Aw TW, Wierzbicka G, Jones DP. Oral glutathione increases tissue glutathione in vivo. Chem Biol Interact. 1991; 80:89-97.
90) Lash LH, Hagen TM, Jones DP. Exogenous glutathione protects intestinal epithelial cells from oxidative injury. Proc Natl Acad Sci USA. 1986; 83:4641-4645.
91) Liposomal formulation for oral administration of glutathione (reduced): United States Patent 20060099244, http://www.freepatentsonline.com/20060099244.html
92) Novi AM. Regression of a.atoxin B1-induced hepatocellular carcinomas by reduced glutathione. Science. 1981; 212:541-542.
93) Palamara AT, Perno C-F, Ciriolo MR, et al. Evidence for antiviral activity of glutathione: in vitro inhibition of herpes simplex virus type 1 replication. Antiviral Res. 1995; 27:237-253.
94) Exner R, Wessner B, Manhart N, Roth E. Therapeutic potential of glutathione. Wien Klin Wochenschr. 2000; 112:610-616.
95) Ibid., 94, pp 610-616
96) Bounous G., Gold P. Anti-cancer therapeutic compositions containing whey protein concentrate. United States Patent 5,888,552 March 30, 1999 , http://www3.sympatico.ca/johnfk/patents.htm
97) Burk RF, ed. Selenium in Biology and Human Health. New York, NY: Springer-Verlag; 1994.
98) Olmsted L, Schrauzer GN, Flores-Arce M, Dowd J. Selenium supplementation of symptomatic human immunode.ciency virus infected patients. Biol Trace Elem Res. 1989; 20:59-65.
99) Ip C. Lessons from basic research in selenium and cancer prevention. J Nutr. 1998; 128:1845-1854.
100) Huttunen JK. Selenium and cardiovascular diseases –an update. Biomed Environ Sci. 1997; 10:220-226.
101) Reilly C. Selenium: a new entrant into the functional food arena. Trends Food Sci Technol. 1998; 9:114-118.
102) Scott R, MacPherson A, Yates RWS, et al. The effect of oral selenium supplementation on human sperm motility. J Urol. 1998; 82:76-80.
103) Dworkin BM. Selenium de.ciency in HIV infection and the acquried immunode.ciency syndrome (AIDS). Chem Biol Interact. 1994; 91:181-186.
104) Alaejos MS, Romero FJD, Romero CD. Selenium and cancer: some nutritional aspects. Nutrition. 2000; 16:376-383
105) Suadicani P, Hein HO, Gyntelberg F. Serum selenium concentration and risk of ischaemic heart disease in a prospective cohort study of 3,000 males. Atherosclerosis. 1992; 96:33-42.
106) Patrick L. Toxic metals and antioxidants: part II the role of antioxidants in arsenic and cadmium toxicity -Toxic Metals Part II, Alternative Medicine Review, May, 2003
107) Schrauzer GN. Selenomethionine: a review of its nutritional signi.cance, metabolism and toxicity. J Nutr. 2000; 130:1653-1656.
108) Eades M. Doctors Complete Guide Vitamins Minerals, page 496, 1994
109) Roberfroid MB, Delzenne NM. Dietary fructans. Annu Rev Nutr. 1998; 18:117-143.
110) Menne E, Guggenbuhl N, Roberfroid M. Fn-type chicory inulin hydrolysate has a prebiotic effect in humans. J Nutr. 2000; 130:1197-1199.
111) Roberfroid MB, Van Loo JA, Gibson GR. The bi.dogenic nature of chicory inulin and its hydrolysis products. J Nutr. 1998; 128:11-19.
112) Williams CM. Effects of inulin on lipid parameters in humans. J Nutr. 1999; 129(7 Suppl):1471S-1473S.
113) Reddy BS, Hamid R, Rao CV. Effect of dietary oligofructose and inulin on colonic preneoplastic aberrant crypt foci inhibition. Carcinogenesis. 1997; 18:1371-1374.
114)Meyer, P. Nondigestible Oligosaccharides as Dietary Fiber, Journal of AOAC International, Volume 87, Issue 3, May 2004, pp: 718-726
115) Abrams S, Grif.n I, Hawthorne K, Liang L, Gunn S, Darlington G, Ellis K (2005). A combination of prebiotic short-and long-chain inulin-type fructans enhances calcium absorption and bone
mineralization in young adolescents. Am J Clin Nutr 82 (2): 471-6. PMID 16087995.
116) Coudray C, Demigné C, Rayssiguier Y (2003). Effects of dietary .bers on magnesium absorption in animals and humans. J Nutr 133 (1): 1-4. PMID 12514257.
117) Wong A, Toth I. Lipid, sugar and liposaccharide based delivery systems. Current Medicinal Chemistry 8(9):1123-1136 (2001).
118) Lasic DD, et al. Spontaneous vesiculation. Advances in Colloid and Interface Science 89-90:337-349 (2001).
119) Igarashi A., et al. Liposomal photofrin enhances therapeutic ef.cacy of photodynamic therapy against the human gastric cancer. Toxicology Letters 145(2):133-141 (2003).
120) Rivera E. Liposomal anthracyclines in metastatic breast cancer: clinical update. Oncologist 8(Suppl.2):3-9 (2003).
121) Justo OR, Moraes AM. Incorporation of antibiotics in liposomes designed for tuberculosis therapy by inhalation. Drug Delivery 10(3):201-207 (2003).
122) Steele, G, Jr, et al. Speci.c active immunotherapy with butanol-extracted, tumor-associated antigens incorporated into liposomes. Surgery 96(2):352-359 (1984).
123) Lopez-Berestein G, et al. Prophylaxis of Candida albicans infection in neutropenic mice with liposome-encapsulated amphotericin B. Antimicrobial Agents and Chemotherapy 25(3):366-367
124) Chaize B, et al. Encapsulation of enzymes in liposomes: high encapsulation ef.ciency and control of substrate permeability. Arti.cial Cells, Blood Substitutes, and Immobilization Technology
125) Sato H, et al. Enhancement of the intestinal absorption of a cyclosporine derivative by milk fat globule membrane. Biological and Pharmaceutical Bulletin 17(11):1526-1528 (1994).