Shark Cartilage: Research Proving the Absorption of Large Polypeptide Proteins Supports Oral Use

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Shark Cartilage: Research Proving the Absorption of Large Polypeptide Proteins Supports Oral Use

Abstract

Although there is growing scientific evidence to the contrary, it is frequently stated that the angiogenic inhibiting proteins identified in shark cartilage are not effective when taken orally. Critics argue that the proteins are broken down in the stomach and are too large to be absorbed in the small intestines. A review of the current literature reveals multiple researchers agreeing that partial intact intestinal absorption occurs with polypeptide proteins up to as large as 80,000 Da (daltons) in molecular weight.( 1) This evidence proves the partial intact absorption of the anti-angiogenic polypeptide fractions in shark cartilage with molecular weights ranging from 1,0000 to 10,000-30,000 Da.( 2) The data shows a logarithmic relationship between molecular weight and intestinal absorption.( 3) The smaller the molecular weight the greater the absorption and the higher up in the small intestines the greater the absorption. Research has also verified that intact proteins survive the hydrochloric acid (HCl) produced in the stomach.( 4) Gastrointestinal tract pore sizes vary greatly as well, further influencing intact absorption of the larger polypeptides.( 5)

Introduction

Since the major arguments challenging the efficacy of the oral administration of shark cartilage are based on questioning intestinal absorption, this paper scientifically reviews the large body of literature available on the intact absorption of polypeptide proteins. It is immanently clear that most researchers specializing in polypeptide absorption agree that large polypeptides of the size in shark cartilage do get partially absorbed intact through the small intestines. The variables effecting this absorption are both fascinating and numerous. They are discussed at length in the sections below.

Method

Our investigation into the absorption of shark cartilage begins with a review of the research evaluating the anti-angiogenic activity of shark cartilage and comparing it to other forms of cartilage. The literature reviewed isolating the protein polypeptide fractions found in shark cartilage with molecular weight determinations.

The paper continues with an evaluation of the extensive body of literature investigating intact polypeptides absorption. The many variables effecting absorption are considered including hydrochloric acid (HCl) production in the stomach, substance density and time of ingestion, the size of the polypeptides, the effect of location in the small intestines on intestinal absorption, the variability of intestinal pore sizes, varied absorption from individual to individual and varied absorption due to certain disease states. Conclusions are drawn based on the evidence presented by multiple researchers in their respective areas of inquiry.

Background Literature on Angiogenic Inhibition

In order for shark cartilage to be an effective oral agent in cancer and other angiogenic mediated diseases it must be proven that it functions as a relatively potent angiogenic inhibitor and that the angiogenic inhibiting polypeptides are absorbed intact through the small intestines. Many researchers, including I. William Lane, PhD and others have shown shark cartilage's ability to inhibit angiogenesis in vitro as well as in vivo on pathological slides of cancer patients in a clinical trial performed in Cuba beginning in June of 1992.( 6-12) Lee and Langer in 1983 determined that shark cartilage contains approximately 1,000 times more anti-angiogenic factors than bovine cartilage. Lee and Langer also determined that sharks are 6% cartilage compared to cattle which are 0.5%.( 13)

Most authors agree that angiogenesis plays a significant role in development of cancer and anti-angiogenic agents are promising anti-cancer therapies.( 14-25) In 1990 Oikawa further confirmed the antiangiogenic effects of shark cartilage.

"Guanidine extraction and crude fractionation of Japanese shark cartilage by ultrafiltration on a molecular weight basis were conducted and the antiangiogenic activities were assayed as to the inhibitions of tumor and embryonic angiogenesis. Significant inhibition of angiogenesis was found and there was a linear relationship between the results of the two assays."( 26)

The ultimate test of intact angiogenic inhibiting polypeptide absorption is the ability of these proteins to inhibit cancer cells in vivo. Research from the Institu Jules, Bordet, Brussels in 1989 with nude mice shows that when shark cartilage was administered daily by mouth at the same time as the subcutaneous introduction of cancer cells, a 50% reduction in tumor size was noted versus the controls.( 27) The study was repeated with a similar reduction in tumor size.( 28) Just recently J.R. Lott, PhD at the University of North Texas completed similar research on mice with compromised immune systems. Thirty balb-C mice, five weeks old, were fed 32 mg/day of shark or bovine cartilage. Lymphosarcoma tumors were implanted into the hip of these mice. Pathological slides were taken from the sarcomas after the day 24-26 of administration with the following results.( 29) Product

Pathologic Slide Result( 29) Benefin shark cartilage

Marked tumor necrosis Cartilade shark cartilage

Slight tumor necrosis Bovine tracheal cartilage

No tumor necrosis

Again, these mice were fed the shark cartilage orally with undeniable tumor inhibiting results.

The Size of Shark Cartilage Polypeptides

Oikawa's research also determined that the molecular weight of the angiogenic inhibiting polypeptides he isolated in shark cartilage are significantly smaller than those in bovine cartilage they are also smaller than those found in all other mammalian sources including bovine cartilage.( 30)

Oikawa's used rabbit corneas for tumor angiogenesis determination and chick embryo choriallantoic mem-branes (CAM) for embryonic angiogenesis determination. "It was found that the inhibitory activities of crude fractions (isolated in shark cartilage) in the molecular weight range of 10( 3) and 10( 4) (filtered through a 10( 3) and/or 10( 4) Da cut-off, PSAC millipore membrane detected with the two assays were correlated with each other, and were not influenced by heat treatment."( 31) The Japanese shark cartilage (glyphis glaucus) was extracted and separated into four fractions of the size described above. Angiogenic inhibition was detected in these crude fraction of molecular weight between 10( 3) and 10( 4) ( 1,000 to 10,000 daltons).

Furthermore, Oikawa writes that when Lee and Lagner performed guanidine extraction for cartilage of the shark, they found that it contained a larger amount of cartilage than the calf. They compared the crude extract with that of calf cartilage by SDS electrophoresis, the patterns of which were different, particularly in the molecular weight range of 2 x 10( 4) ( 20,000 daltons). He goes on to say that the angiogenic inhibitors isolated from shark cartilage are smaller than that of calf cartilage and smaller than what Lee and Langer isolated and are heat stable.( 32)

Four years earlier in 1986, Luer prepared crude protein extracts from the cartilage of the bull shark (carcharhinus leucas) and the sandbar shark (carcharhinum plumbeus) by exposure of the cartilage to a similar process using guanidine chlorides and ultrafiltration. The protein fractions were then isolated for their ability to inhibit protease activity and the neovascularization associated with the process of angiogenesis. "Maximum inhibition of trypsin, chymotrysin and pronase is associated with protein fractions falling in the molecular weight range of 13,000 to 18,000 daltons, while fractions in the range of 25,000 to 33,000 daltons result in the greatest inhibition of collagenase activity and of the neovascular spread of skate embryo yolk sac blood vessels."33 However, this research was only published as an abstract in Federal Proc. It was conducted privately by Mote Marine Laboratory without any published detailed experimental design.

Intestinal Absorption Controversy Resolved

The therapeutic efficacy controversy concerning shark cartilage has mostly centered around the question of whether or not anti-angiogenic compounds get absorbed through the intestinal tract intact. Research described above has shown that the anti-angiogenic compounds are large polypeptide structures. Conventional theory holds that the size of these proteins are too large to be absorbed intact through the gastrointestinal tract.

However, there is a new paradigm of understanding concerning intact absorption of large polypeptides that has been verified by multiple researchers from varying perspectives.

One of these perspectives is offered by Michael Murray, ND, respected by his peers as a fair and innovate researcher and reviewer of current literature. He stated that;

"Contrary to long held theories that the healthy individual does not absorb proteins and large polypeptides intact, there is now irrefutable evidence that many large macromolecules are absorbed intact from the human gut into the bloodstream under normal conditions.( 30) In some instances, the body appears to recognize which molecules it needs to absorb intact and which molecules it needs to break down into smaller units. This phenomenon may help to explain the effectiveness of glandular therapy."( 34)

Gardner, in his leading article, who was references by Murray in this former quote elaborates by saying; "There now is irrefutable evidence that small amounts of intact peptides and proteins do enter the circulation under normal circumstances....There is now no reasonable doubt that small quantities of intact proteins do cross the gastrointestinal tract in an im nis and adult humans, and that this is a physiologically normal process required for antigen sampling by subepithelial immune tissue in the gut....The process of intact protein absorption occurs without eliciting harmful consequences for most individuals, but it appears likely that a small number of people absorbing these "normal" amounts may react idiosyncratically; also, some individuals may absorb excess amounts, and they may suffer clinically significant consequences."( 35)

Over 50,000 patients worldwide have taken shark cartilage over the last five years. No adverse immune binding or allergic reactions to shark cartilage have been reported in the literature or by any treating physicians.

As early as 1980 Mathews and Payne in their chapter titled Transmembrane Transport of Small Peptides in Current Topics in Membranes and Transport, Vol 14, admitted that "not long ago, the author of this section (Mathews) might have written that it was probable that there was no significant mediated uptake of tetra or higher peptides, but a recent report has provided evidence for uptake by the absorptive cells of at least one tetrapeptide...."( 36)

What Molecular Weights Can Pass Through the Intestinal Tract Intact?

The question then becomes, what is the largest molecular size that can pass through the intestinal tract intact? Loehry, CA, et.al. in 1970 writes that albumin and polyvinylpyrrolidone (PVP) are known to pass from the body into the small intestinal lumen and the above experiments confirm that PVP with an effective molecular weight of 33,000 can pass both from the intestinal lumen into the plasma and in the opposite direction.( 37) He postulates that pores with a much greater diameter than 9 angstrom exist or alternatively that a different mechanism of permeation has been studied. He showed that there is a linear relationship between log molecular weight and log clearance. See figure 1. This relationship holds for molecular weights between 60 and 33,000 as well as 8,000 to 80,000. In other words, the larger the molecule the less absorption (intestinal clearance).

These findings suggest that the permeating mechanism is the same for the large molecules as for small ones, otherwise one might expect to see a cut-off phenomenon above a certain molecular size.( 38) If the pore hypothesis is correct, the data could be accounted for by postulating a normal distribution of pore size, i.e., a very few large pores, a moderate number of medium-sized pores, and very many small pores. The gastrointestinal mucosal wall was thought to be limited by a fat soluble membrane with water-filled pores in the range of 3-9 angstrom in diameter. Yet substances cross the capillary wall through pores in the fenestra at a diameter from 70-500 angstrom. These substances then either pass out between mucosal cells or at the tip of villi through the gap left by exfoliated cells or through the mucosal cells themselves, leaking form the epithelial border back into the bowel lumen.( 39)

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Chadwich, VS, et.al. further substantiated Loehry's work seven years later and found that by using polyethylene glycol (PEG 400) showed that permeability for larger molecules diminishes from proximal to distal intestines.( 41) Numerous animal studies with rats, rabbits and fish have verified intestinal absorption of polypeptides in a molecular weight ranging from 20,000 to 50,000.( 42)

Discussion

Hydrochloric acid (HCl) production in the stomach further impairs optimal intact polypeptide absorption in the duodenum and lower down the small intestines. HCI begins some of the cleavage of the polypeptides before entry into the small intestines. In spite of this, intact small intestines absorption of polypeptides occurs.( 42) This is especially true for compounds ingested as liquids on an empty stomach. These compounds pass through the stomach more quickly than solids mixed with a meal.( 43) The percentage of absorption would likely be increased if practitioners prescribed the administering of sodium bicarbonate 5-15 minutes before taking shark cartilage on an empty stomach. This would neutralize the acid stomach pH caused by hydrochloric acid production and further enhance intact polypeptide absorption of shark cartilage in the small intestines. It is essential to wait 30 minutes or more after the bicarbonate and shark cartilage dosage before eating to allow the pH of the stomach to return to normal. This will optimize protein digestion with each meal.

The degree of intestinal absorption varies to some extent from individual to individual, regionally throughout the small intestines and with certain diseased states.( 44) However, the absorption of intact polypeptides occurs normally with healthy individuals as well.( 45) Patients with conditions such as leaky gut syndrome, food allergies, tropical sprue (gluten enteropathy), Crohn's disease (inflammation at the end of the small intestines) will likely absorb greater amounts of intact polypeptides.( 46)

Critics have used the analogy of insulin not being absorbed oral to defend their argument refuting shark cartilage polypeptide absorption. Insulin has a molecular weight of 6,000 with negligible additional weight ( 50-150) added by small amounts of heavy metals such as cadmium, cobalt, nickel or zinc as incorporated components of the polypeptide molecule.( 47) It is clearly apparent from this study that some insulin absorption occurs through an oral route. However, in the control of diabetes varied absorption from individual to individual as well as varied absorption at different regions of the gastro-intestinal tract makes controlled regulation of blood sugar difficult.

Shark cartilage has some of its polypeptides at smaller molecular weights of 1,000 compared to the 6,000 of insulin. A relatively higher percentage of these polypeptides will get absorbed orally than that of insulin.

It has been well established in biological terrain medicine that increased acidity in saliva increased one's level of chronic pain.( 48) Adding bicarbonate to treatment protocols for shark cartilage is likely to further reduce the level of chronic pain in cancer and arthritis patients using shark cartilage.

The large doses of calcium in shark cartilage are natural buffers reducing acidity and increase intact intestinal absorption. This reduction of acidity is an additional mechanism explaining how shark cartilage reduces pain in arthritis and cancer in addition to its anti-inflammatory properties due to muco-polysaccharide content.

Conclusion

Intact Polypeptide absorption occurs most readily:

- with polypeptides of lower molecular wt.

- higher up in the intestinal tract

- in areas of larger intestinal pore size

- with liquids on an empty stomach

- in a more alkaline stomach pH

- in certain disease states

There is now irrefutable evidence that protein polypeptides of the size naturally occurring in shark cartilage do get absorbed to some extent intact in the gastro-intestinal tract. This occurs more readily with polypeptides of lower molecular weight, higher up in the intestinal tract, in areas of larger intestinal pore size, with liquids on an empty stomach, in a more alkaline stomach pH and in certain disease states.

Additional variables effecting absorption include levels of HCl production and whether the product in question is a solid or a liquid, taken on an empty or a full stomach. There is also some variability in absorption from person to person although absorption takes place in the normal individual. Certain disease states increase absorption of intact polypeptides.

Disclosure

This study was funded by Lane Labs, the manufacturer of BeneFin and ImmunoFin with the understanding in advance that the results would be published, regardless of the findings.

Correspondence:

Martin Milner, ND, President

Center for Natural Medicine, Inc.

Diagnostic Consultants & Gift of Life Foundation

Health Outcomes Research Division

1330 S.E. 39th Avenue

Portland, Oregon 97214 USA

503-232-1100

Fax 503- 232-7751

E Mail: martinm@teleport.com

Web Site: http://www.cnm-inc.com

Lane Labs

110 Commerce Drive

Allendale, New Jersey 07401 USA

800-526-3005

Fax 201-236-9090
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(2.) Oikawa, T, et al., A novel angiogenic inhibitor derived from Japanese shark cartilage. Extraction and estimation of inhibitory activities toward tumor and embryonic angiogenesis, Cancer Letters, 1990, 51:181-186.

(3.) Loehry, AC, Axon, ATR, Hilton, PJ, Hider, RC, Creamer B., Permeability of the small intestine to substances of different molecular weight, Gut, 1970, 11, 466-70.

(4.) Gardner, MLG, Gastrointestinal absorption of intact proteins, Ann. Rev. Nutr, 1988, 8:329-50, Gardner, MLG, Passage of intact peptides across the intestine, Adv. Biosci, 65:99-106, Murray, M., Thymus extracts, Journal of Naturopathic Medicine, 1994, 5(1):77-79, Mathews, DW, Payne JW, in Transmembrane Transport of Small Peptides in Current Topics in Membranes and Transport, Vol 14, Academy Press, 1980, Loehry, AC, Axon, ATR, Hilton, PJ, Hider, RC, Creamer B., Permeability of the small intestine to substances of different molecular weight, Gut, 1970, 11, 466-70, Chadwich, VS, Phillips,, SF, Hofmann, AF, Measurements of intestinal permeability using low molecular weight polyethylene glycols (PEG 400), Gastroenterology, 1977, 73:247251, Walker, WA, Wu, M, Isselbacher, KJ, Bloch, KJ, Intestinal uptake of macromolecules, Jr. Immunology, 1975, 115(3) 854-61, Walker, WA, Isselbacher, KJ, Uptake and transport of macromoleules by the intestine: possible role in clinical disorders, Gastroenterology, 1974, 67:531-50, Bjarnason, I, Peters, TJ, Helping the mucosa make sense of macromolecules, Gut, 1987, 28, 1057-61, Hemming, WA, Williams, EW, Transport of large breakdown products of dietary protein through the gut wall, Gut, 1978, 19:715-23.

(5.) Loehry, AC, Axon, ATR, Hilton, PJ, Hider, RC, Creamer B., Permeability of the small intestine to substances of different molecular weight, Gut, 1970, 11, 466-70.

(6.) Lane, I. William, Shark cartilage therapy results and research today, Physician Information Package, Cartilage Consultants, spring 1995.

(7.) Folkman, Brem, Inhibition of tumor angiogenesis mediated by cartilage, J. Exp. Med, 141:p427-439.

(8.) Luer, C., Inhibitors of Angiogenesis from Shark Cartilage (Meeting Abstract), Mote Marine Lab., Fed. Proc. 1986, 45(4):949.

(9.) Moses, MA, etal, Identification of an inhibitor of neovascularization from cartilage, Science, 1990, 248:630-634.

(10.) Oikawa, T, et al., A novel angiogenic inhibitor derived from Japanese shark cartilage. Extraction and estimation of inhibitory activities toward tumor and embryonic angiogenesis, Cancer Letters, 1990, 51:181-186.

(11.) Lane, I.W., Komak, L., Sharks Don't Get Cancer, Avery, 1992.

(12.) Britto, Fernandez J., Lane, I.W., Angiogenesis modulation in peritumor (or connective tissue by cartilage from shark, The Cuban experience, XVII World Congress of Anatomic and Clinical Pathology, 1993, Mexico.

(13.) Lee, A., Langer R., Shark cartilage contains inhibitors of tumor angiogenesis, Science, Sept. 1983, 221: 1185-87.

(14.) Blumberg, N., Tumor angiogenesis factor. Speculations on an approach to cancer therapy. Yale J. of Biol. and Med., 1974,47:71-81.

(15.) Folkman, J., Tumor angiogenesis: A possible control point in tumor growth, Ann. Int. Med., 1975, 82:96-100.

(16.) Folkman, J., The vascularization of tumors, Sci. Am., May 1976, 234(5):58-64, 70-73.

(17.) Langer, R., etal, Isolation of a cartilage factor that inhibits tumor neovascularization, Science, Jul. 1976, 193(4247):70-72.

(18.) Sadove, AM, etal, Inhibition of mammary carcinoma invasiveness with cartilage-derived inhibitors, Surg Forum Orthopaed Surg, 1977, 28:499501.

(19.) Pauli, B.U., etal, Regulation of tumor invasion by cartilage-derived anti-invasion factor in vitro, J. Natl. Can. Inst., July 1981, 67:65-73.

(20.) Folkman, J., How blood is regulated in normal and neoplastic tissue, Cancer Res., Feb. 1986, 48:467-473.

(21.) Folkman, J, Klagnbrun, M., Angiogenic factors, Science, Jan. 1987, 235:235-247.

(22.) D'Amore, P.A., Anti-angiogenesis as a strategy for anti-metastasis, Seminars in Thrombosis & Hemostasis, 1988, 14(1):73-78.

(23.) Folkman, etal., Induction of angiogenesis during the transition from hyperplasia to neoplasia, Nature, 1989, 339(6219):L58-61.

(24.) Weidher, N., etal., Tumor angiogenesis and metastasis correlation in invasive breast carcinoma, New Eng J Med, Jan. 3, 1991, 324:1-8.

(25.) Wilson, J., Shark Cartilage: A review of background literature & research, Townsend Letter for Doctors, Aug/Sept 1994, 133-134:864872.

(26.) Oikawa, T, etal, A novel angiogenic inhibitor derived from Japanese shark cartilage. Extraction and estimation of inhibitory activities toward tumor and embryonic angiogenesis, Cancer Letters, 1990, 51:181-186.

(27.) Institut Jules Bordet, Brussels, Effect of cartilage administration against a human melanoma xenograft (MeXF 514) in nude mice, Comprehensive Research Papers on the Therapeutic Use of Shark Cartilage, Lane, IW, 1989 Institut research date.

(28.) Ibid.

(29.) Lott, JR, N. Texas University, current research about to be published from telephone conversations with IW Lane.

(30.) Oikawa, T, etal., A novel angiogenic inhibitor derived from Japanese shark cartilage. Extraction and estimation of inhibitory activities toward tumor and embryonic angiogenesis, Cancer Letters, 1990, 51:181-186.

(31.) Ibid.

(32.) Ibid.

(33.) Luer, C., Inhibitors of Angiogenesis from Shark Cartilage (Meeting Abstract), Mote Marine Lab., Fed. Proc. 1986, 45(4):949.

(34.) Gardener, M.L.G., Gastrointestinal absorption of intact proteins, Ann Rev Nutr, 1988, 8:329-350.

(35.) Murray, M., Thymus extracts, Journal of Naturopathic Medicine, 1994, 5(1):77-79.

(36.) Gardener, M.L.G., Gastrointestinal absorption of intact proteins, Ann Rev Nutr, 1988, 8:329-350.

(37.) Mathews, DW, Payne JW, in Transmembrane Transport of Small Peptides in Current Topics in Membranes and Transport, Vol 14, Academy Press, 1980.

(38.) Loehry, AC, Axon, ATR, Hilton, PJ, Hider, RC, Creamer B., Permeability of the small intestine to substances of different molecular weight, Gut, 1970, 11, 466-70.

(39.) Ibid.

(40.) Ibid.

(41.) Ibid.

(42.) Chadwick, VS, Phillips,, SF, Hofmann, AF, Measurements of intestinal permeability using low molecular weight polyethylene glycols (PEG 400), Gastroenterology, 1977, 73:247-251.

(43.) Udall, JN, Colony, P, Pang, FK, Trier, JS, Walker, WA, Development of gastrointestinal mucosa barrier II. The effect of natural vers artifical feeding on intestinal permeability to macromolecules, Pediatric Res. 1981, 15:245-49, Walker, WA, Wu, M, Isselbacher, KJ, Bloch, KJ, Intestinal uptake of macromolecules, Jr. Immunology, 1975, 115(3) 854-61, Block, KJ, Bloch, D, Stearns, M, Walker, W, Intestinal uptake of macromoledules, VI. Uptake of protein angtigen in vivo in normal rats and in rats infected with Niippostronglus brasiliensis or subjected to mild systemic anaphylaxis. Gastroenterology, 1979, 77:1039-44, Ducros, R, Heyman, M, Beaufrere, B, Morgat J, Desjeux J, Horseradish peroxidase transport across rabbit jejunum and Peyer's patches in vitro, Am J. Physiol., 1983, 245:G54-G58, Hamilton, I, Rothwell J, Archer, D, Axon, ATR, Permeability of the rat small intestine to carbohydrate probe molecules, Clin. Sci., 1987, 73:189-196, Hemmings C, Hemmings WA, Patey RL, Wood, C, The ingestion of dietary protein as large molecular mass degradation products in adult rats, Proc. R. Soc. London Ser. B, 1977, 198:439-53, Limm, PL, Rowley, D, The effect of antibody on the intestinal absorption of macromolecules and on intestinal permeability in adult mice, Int. Arch. Allergy Appl. Immunol., 1985, 76:30-36, McLean, E, Ash, R, The timecourse of appearance and net accumulation of horseradish peroidase (HRP) presented orally to rainbow trout, Salmongairdneri (Richardson) Comp. Biochem. Physiol, 1987, 88A:507-10, McLean, E, Ash, R, Intact protein (antigen) absorption in fishes: mechanism and physiological significance, J. Fish Biol., 1987, 31 (Suppl. A):219-23.

(44.) Ibid.

(45.) Netter, F., The Ciba Collection of Medical Illustrations, Vol. 3, Digestion, Factors influencing gastric activity, slide 853.

(46.) Gardener, M.L.G., Gastrointestinal absorption of intact proteins, Ann Rev Nutr, 1988, 8:329-350.

(47.) Ibid.

(48.) Ibid., Bjarnason, I, Marsh, MN, Levi, AJ, Crow, T, Goolamali, S, et al, Intestinal permeability in cellar sprue (CS) dermatitis herpetiformis (DH), schizo-phrenia (S) and atopic eczema (AE), Gastroenterology, 1984, 86:1029, Bjarnason, Et.al, A persistent defect in intestinal permeability in coeliac disease revealed by a 51 Cr-labelled EDTA absorption test, Lancet, 1:323-5, Jackson, PG, Lessof, MH, et.al, Intestinal permeability in patients with eczema and food allergy, Lancet, 1981, 1:1285-86.

(49.) Merck Index, 11th edition, p4887.

(50.) Vincent, LC, Bio-electronogram, adapted in Understanding Biological Terrain and Its Overall Significance, Professional Health Products publication for Robert Greenberg, DC's seminar, Kemeny, co-vice Chancellor of Biomathematics at the Polytechinic School in Budapest, 1953, research on biological terrain measurement, Understanding Biological Terrain and Its Overall Significance, Professional Health Products publication for Robert Greenberg, DC's seminar.

Townsend Letter for Doctors & Patients.

~~~~~~~~

By Martin Milner

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