Modified Citrus Pectin & Galectin-3 The Role of Modified Citrus Pectin and Galectin-3 in the Prevention and Treatment of Metastatic Cancer

Galectin-3 & Disease Elevated Galectin-3 in the body is directly involved in: Cancer Cardiovascular Disease Chronic Hepatitis Kidney Disease Diabetes Inflammation & Fibrosis Others

Galectin-3 Levels & Mortality from All Causes in the General Population: PREVEND Overall Average 11.9

Galectin-3 & Cancer In Cancer, Galectin-3 plays a role in: Cell to Cell Adhesion Aggregation of Cancer Cells Tumor Growth Metastasis Angiogenesis Inhibition of Apoptosis

Blood Vessel

Blood Vessel

What is Modified Citrus Pectin (MCP) ? MCP is derived from the pith of citrus fruit peels Complex polysaccharide fiber of repeating galacturonic acid groups with neutral sugar side chains Unmodified citrus pectin molecular weight 50 to 300 kiloDaltons (kDa) with esterification ~70% Optimal biological activity: molecular weight <13 kDa with esterification <10%, and specific structure

Modified Citrus Pectin is a Galectin-3 Blocker Binds to Galectin-3 molecules Blocks aggregation of cancer cells Blocks docking of cancer cells Blocks interactions with endothelium necessary for angiogenesis

Modified Citrus Pectin Cancer Research

Benefits of MCP in Cancer Treatment Anti-Cancer and Anti-Metastasis Blocking of Galectin-3 Effects Synergistic Effect with Chemotherapy Protection Against Post-Radiation Damage Improved Quality of Life

Inhibition of Spontaneous Metastasis in a Rat Prostate Model by Oral Administration of Modified Citrus Pectin Pienta KJ, Naik H, Akhtar A, Yamazaki K, Replogle, TS, Lehr J, Donat TL, Tait L, Hogan V, Raz A Wayne State University School of Medicine, Detroit, MI, USA J Natl Cancer Inst 1995 87(5):348-53. Method: MCP’s inhibition of prostate cell adhesion to endothelial cells. 0.1% and 1.0% MCP in rats’ drinking water; controls had plain water Results: Significant reduction in lung metastases -- 50% reduction in 0.1% group; 56% reduction in 1.0% group (P<0.03 & P<0.001).

Inhibition of Spontaneous Metastasis in a Rat Prostate Model by Oral Administration of Modified Citrus Pectin Pienta KJ, Naik H, Akhtar A, Yamazaki K, Replogle, TS, Lehr J, Donat TL, Tait L, Hogan V, Raz A Wayne State University School of Medicine, Detroit, MI, USA J Natl Cancer Inst 1995 87(5):348-53. Conclusion: MCP acted as potent inhibitor of spontaneous prostate carcinoma metastasis.

Inhibition of Human Cancer Cell Growth & Metastasis in Nude Mice by Oral Intake of Modified Citrus Pectin Nangia-Makker P, Hogan V, Honjo Y, Baccarini S, Tait L, Bresalier R, Raz A Wayne State University, School of Medicine, Detroit, MI, USA J Natl Cancer Inst. 2002 94(24):1854-62. Method: MCP’s inhibition of breast & colon cancer progression; MCP’s interaction with Galectin-3 Results: 70.2% Reduction in Breast Tumor Growth Breast Angiogenesis: 66% Reduction Breast to Lung Metastasis: 0% MCP v. 100% Control Colon to Liver Metastasis: 0% MCP v. 60% Control Colon to Lymph Metastasis: 25% MCP v. 100% Control

Inhibition of Human Cancer Cell Growth & Metastasis in Nude Mice by Oral Intake of Modified Citrus Pectin Nangia-Makker P, Hogan V, Honjo Y, Baccarini S, Tait L, Bresalier R, Raz A Wayne State University, School of Medicine, Detroit, MI, USA J Natl Cancer Inst. 2002 94(24): 1854-62. Conclusion: MCP inhibits carbohydrate mediated tumor growth, angiogenesis & metastasis via effects on Galectin-3 function

Modified Citrus Pectin Induces Cytotoxicity of Prostate Cancer Cells in Co-Culture with Human Endothelial Monolayers Weiss T, McCulloch M, Eliaz I Amitabha Medical Clinic & Healing Center, Sebastopol, CA, USA EcoNugenics, Santa Rosa, CA, USA Intl Conference on Diet & The Prevention of Cancer 1999, Tampere, Finland. Method: Human vascular endothelial cell layer & PC-3 prostate cancer cells. Results: Strong tumor cell death response with MCP, compared to controls.

Effect of Modified Citrus Pectin on PSA Doubling Time in Prostate Cancer Patients: A Pilot Clinical Trial Strum S, Scholz M, McDermed J, McCulloch M, Eliaz, I Prostate Oncology Specialist, Marina del Rey, CA, USA Amitabha Medical Clinic & Healing Center, Sebastopol, CA, USA EcoNugenics, Santa Rosa, CA, USA. International Conference on Diet & The Prevention of Cancer 1999, Tampere, Finland. Method: MCP 15 g/day to patients with biochemical relapse post local therapy. PSA Doubling Time (PSADT) evaluated at intervals. Results: MCP significantly increased PSADT in prostate cancer patients.

Pilot Clinical Results: MCP’s Effect on PSA Doubling Time Patient MCP Use (Months) PSADT Change Status Patient 1 5 193% Response Patient 2 6 Patient 3 3 80% Patient 4 >15 55% Patient 5 6% P. Response Patient 6 -7% Stable Disease Patient 7 -69% No Response    

Modified Citrus Pectin Increases the Prostate-Specific Antigen Doubling Time in Men with Prostate Cancer: A Phase II Pilot Study Guess BW, Scholz MC, Strum SB, Lam RY, Johnson HJ, Jennrich RI Healing Touch Oncology, Marina del Rey, CA, USA Prostate Cancer & Prostatic Disease 2003;6(4):301-4. Method: 10 men with biochemical prostate cancer relapse used MCP: 15g daily for 1 year.

Modified Citrus Pectin Increases the Prostate-Specific Antigen Doubling Time in Men with Prostate Cancer: A Phase II Pilot Study Guess BW, Scholz MC, Strum SB, Lam RY, Johnson HJ, Jennrich RI Healing Touch Oncology, Marina del Rey, CA, USA Prostate Cancer & Prostatic Disease 2003;6(4):301-4. Results: MCP significantly increased PSADT in 7 out of the 10 participants (p<0.01).

Phase II Study: PSADT Results After 1 Year 968 % * *

Using Splines to Detect Changes in PSA Doubling Times Guess B, Jennrich R, Johnson H, Redheffer R, Scholz M, Healing Touch Oncology, Marin del Ray, CA, Department of Statistics, UCLA, CA, The Prostate 2003 54:88-95.

Typical Patient Results MCP

Clinical Benefit in Patients with Advanced Solid Tumors Treated with Modified Citrus Pectin Azémar M, Hildenbrand B, Haering B, Heim ME, Unger C Albert-Ludwigs-University in Freiburg, Germany Sonnenberg-Klinik, Bad Sooden-Allendorf, Germany Clinical Medicine: Oncology 2007 1:73–80. Method: MCP 15g daily. Results: 49 patients with advanced solid tumors. 29 evaluated after 2 months -- 21% showed stabilization & improvements in quality of life. One patient w/ metastasized prostate carcinoma showed 50% decrease in PSA, with significant increase in quality of life & decrease in pain.

Clinical Benefit in Patients with Advanced Solid Tumors Treated with Modified Citrus Pectin Azémar M, Hildenbrand B, Haering B, Heim ME, Unger C Albert-Ludwigs-University in Freiburg, Germany Sonnenberg-Klinik, Bad Sooden-Allendorf, Germany Clinical Medicine: Oncology 2007 1:73–80. Conclusion: MCP shows clinical benefits & improvements in quality of life in advanced cancer patients.

Inhibitory Effect of Modified Citrus Pectin on Liver Metastasis in a Mouse Colon Cancer Model Liu HY, Huang ZL, Yang GH, Lu WQ, Yu NR Guangzhou Medical College, Guangzhou, China World J Gastroenterol 2008 14(48): 7386-7391. Method: 5 groups of 15 mice. MCP: 0.0%, 1.0%, 2.5% and 5.0%; and negative control. Colon cancer cells injected into spleen except negative control -- liver metastasis observed after 3 wks. ELISA used to detect Galectin-3. Results: MCP groups: Metastasis 80%, 73.3% & 60%. MCP 0.0%: Metastasis100%.

Conclusion: MCP significantly reduced liver metastasis. Inhibitory Effect of Modified Citrus Pectin on Liver Metastasis in a Mouse Colon Cancer Model Liu HY, Huang ZL, Yang GH, Lu WQ, Yu NR Guangzhou Medical College, Guangzhou, China World J Gastroenterol 2008 14(48): 7386-7391. Conclusion: MCP significantly reduced liver metastasis.

PectaSol-C Modified Citrus Pectin Induces Apoptosis & Inhibition of Proliferation in Human & Mouse Androgen Dependent & Independent Prostate Cancer Cells Yan J, Katz A Columbia University Medical Center, New York, NY, USA Integrative Cancer Therapies 2010 9:197-203. Method: 1% MCP treatment of human prostate cancer cell lines (LNCaP & PC3) and mouse prostate cancer cell lines (CASP2-1 & CASP1-1)

4 day MCP treatment showed cytotoxicity: PectaSol-C Modified Citrus Pectin Induces Apoptosis & Inhibition of Proliferation in Human & Mouse Androgen Dependent & Independent Prostate Cancer Cells Yan J, Katz A Columbia University Medical Center, New York, NY, USA Integrative Cancer Therapies 2010 9:197-203. Results: Confirmed apoptosis and inhibition of cancer cell proliferation 4 day MCP treatment showed cytotoxicity: 52.28% in LNCaP 48.16% in PC3 23.03% in CASP2-1 49.01% in CASP1-1

Combination Effect of PectaSol and Doxorubicin on Viability, Cell Cycle Arrest and Apoptosis in DU-145 and LNCaP Prostate Cancer Cell Lines Najmeh T, Houri S, Parvin M, Firouzeh B, Arash HN, Abdolfattah S, Ebrahim H Tehran University, Tehran, Iran Cell Biology International (2012) doi:10.1042/CBI20110309. Method: 48 hour effects of PectaSol on Doxorubicin (Dox) cytotoxicity, apoptosis and cell cycle in prostate cancer cell lines. IC50 Decrease: 1.5 fold IC50 Decrease: 1.3 fold

Combination Effect of PectaSol and Doxorubicin on Viability, Cell Cycle Arrest and Apoptosis in DU-145 and LNCaP Prostate Cancer Cell Lines Najmeh T, Houri S, Parvin M, Firouzeh B, Arash HN, Abdolfattah S, Ebrahim H Tehran University, Tehran, Iran Cell Biology International (2012) doi:10.1042/CBI20110309. Conclusion: Lower, less toxic doses Dox needed when combined with PectaSol.

MCP During Chemotherapy & Radiation MCP can enhance therapeutic effect of chemotherapy drugs and treatment of chemo resistant cancers Cisplatin (Platinol), Bortezomib (Velcade), Dexamethasone (Decadron), Doxorubicin MCP use very important in preventing post chemotherapy & radiation damage Specifically post radiation induced inflammation and fibrosis

Summary: MCP in Cancer Treatment MCP Reduces: Primary Tumor Angiogenesis Metastatic Process MCP Provides: Blocking of Galectin-3 Effects Synergistic Effect with Chemotherapy Protection Against Post-Radiation Damage Improved Quality of Life

Modified Citrus Pectin Immune Research

MCP Immune System Benefits Modified Citrus Pectin: Induces NK Cell Activation Induces NK Cell Activity Activates T Cytotoxic Cells Increases B Cell Activation

Activation of Human T-Cytotoxic Cell, B-Cell, and Natural Killer (NK)-Cells and Induction of NK-Cell Activity Against K562 Chronic Myeloid Leukemia Cells with Modified Citrus Pectin Ramachandran C, Wilk, B, Hotchkiss, A, Eliaz, I & Melnick SJ Dharma Biomedical, Miami, FL, USA, EcoNugenics, Santa Rosa, CA, USA Eastern Regional Research Center, ARS, USDA, Wyndmoor, PA, USA Department of Pathology, Miami Children's Hospital, Miami, FL, USA BMC Complementary and Alternative Medicine 2011, 11:59. Method (Part I): Healthy human blood samples incubated with increasing doses of MCP and antibodies. At 24 hours samples lyzed and run on a flow cytometer. Analyzed % of activated T-cytotoxic cell, B-cells, and NK-cells; and % increase over untreated control.

Results Part I: MCP Activates T-Cytotoxic Cells ** * *

Results Part I: MCP Increases B-Cell Activation *** **

Results Part I: MCP Increases NK Cell Activation ** ** * *

MCP: Induction of NK-Cell Activity Against K562 Chronic Myeloid Leukemia Cells Method (Part II): NK cell ability to induce leukemia cell death analyzed by co-incubating MCP-treated lymphocytes with K562 T-cell leukemia cells. Healthy human lymphocyte samples treated with increasing doses of MCP. After 24 hours, K562 labeled. Plates returned to incubator (for 4 hrs) to induce leukemia cell death.

Results Part II: MCP Induces NK Cell Activity

Modified Citrus Pectin: Chelation & Detoxification Research

MCP Heavy Metal Elimination Forms stacked “egg box” structure Each pocket negatively charged Negative charge binds heavy metals Toxic metal trapped in the “egg box” Safely excreted from the body

The Effect of Modified Citrus Pectin on Urinary Excretion of Toxic Elements Eliaz I, Hotchkiss AT, Fishman ML, Rode D; Amitabha Medical Clinic & Healing Center, Sebastopol, CA, USA; Eastern Regional Research Center, ARS, USDA, Wyndmoor, PA, USA; UC, Davis, CA, USA Phytother Res 2006 20(10):859-64. Methods: MCP administered for 6 days. Baseline 24 hr urine collection before MCP, and on days 1 and 6. Days 1 - 5: MCP 15 g/day & Day 6: MCP 20 g/day.

The Effect of Modified Citrus Pectin on Urinary Excretion of Toxic Elements Eliaz I, Hotchkiss AT, Fishman ML, Rode D; Amitabha Medical Clinic & Healing Center, Sebastopol, CA, USA; Eastern Regional Research Center, ARS, USDA, Wyndmoor, PA, USA; Univer. of CA, Davis, CA, USA Phytother Res 2006 20(10):859-64. Results: Urinary excretion of lead, mercury, cadmium & arsenic increased. Essential minerals not changed. No side effects reported.

The Effect of Modified Citrus Pectin on Urinary Excretion of Toxic Elements # * p < .05 # p < .10 # # * * * #

MCP & Urinary Excretion of Toxins MCP Chelation: Increased urinary excretion of toxic metals Demonstrated heavy metal chelation due to reduced molecular size & esterification 10% rhamnogalacturonan II -- known for binding affinity and immune enhancement Does not affect essential minerals No side effects reported

Modified Citrus Pectin Decreases the Total Body Burden: A Pilot Human Clinical trial Eliaz I. Amitabha Medical Clinic & Healing Center, Sebastopol, California, USA. EcoNugenics, Santa Rosa, California, USA. 228th ACS National Meeting, Philadelphia, PA. 2004. Methods: Oral intake 5 g Modified Citrus Pectin/3x day for 4-10 months. Base line body burden and change measured with DMPS challenge (250mg i.v. followed by 6 hr. urine collection). Results: All subjects showed significant decrease in Mercury levels. Average decrease was 62.17%, ranging between 38.13% & 74.83% (p=0.0313). No significant side effects were noted.

Study Conclusion Percent Reduction in Mercury from Baseline MCP was effective in decreasing the total body burden of Mercury in all subjects. MCP is a promising systemic gentle chelator of heavy metals that can be used on an on going basis. 10 months 4.5 months 4 months 6 months 6.5 months MCP Intervention Individual Results

The Role of Modified Citrus Pectin as an Effective Chelator of Lead in Children Hospitalized with Toxic Lead Levels Zhao ZY, Liang L, Fan X, Yu Z, Hotchkiss AT, Wilk BJ, Eliaz I Children’s Hospital, Zhejiang University, School of Medicine, Hangzhou, Republic of China Centrax International, Inc, San Francisco, CA, USA Eastern Regional Research Center, ARS, USDA, Wyndmoor, PA, USA EcoNugenics, Santa Rosa, CA, USA Altern Ther Health Med. 2008 14(4):34-8.

Lead in Blood Serum P Value = 0.0016

Lead in 24 Hour Urine Excretion P Value = 0.0007

Synergistic Benefits of MCP

Prostate Poly Botanical Formula Integrative blend of 33 ingredients: Vitamins, Minerals, Botanically Enhanced Medicinal Mushrooms, and Botanical Extracts

ProstaCaid Induces G2/M Cell Cycle Arrest & Apoptosis in Human & Mouse Androgen-Dependent & -Independent Prostate Cancer Cells Yan J & Katz AE; Department of Urology, Columbia University Medical Center, New York, NY, USA. Integr Cancer Ther 2010 9:186-196. Effects on cell viability on androgen-dependent, LNCaP (A) & CASP 2.1 (C), & androgen-independent PC3 (B) & CASP 1.1 (D) Effects on long-term cell viability by colony formation

Effect of ProstaCaid on Invasive Behavior of Prostate Cancer Cells (A) Cell adhesion. PC-3 cells were treated with ProstaCaid for 24 hours and cell adhesion to fibronectin determined. (B) Cell migration. Cell migration was determined after 24 hours of incubation with ProstaCaid in Boyden Chambers. C D uPA Density 1.00 0.61 0.38 0.16 uPA 0 20 40 80 ProstaCaid ( mg/ml) (D) uPA secretion. PC-3 cells were treated for 24 hours and the expression of uPA detected in conditioned media with anti-uPA antibody by Western blot. * p<0.05 (C) Cell invasion. Cell invasion was determined after 24 hours of incubation with ProstaCaid in Boyden Chambers coated with Matrigel.

ProstaCaid Inhibits Tumor Growth in a Xenograft Model of Human Prostate Cancer Results: No effect on body weight or activity of liver enzymes (ALT, AST). No sign of toxicity in liver, spleen, kidney, lung and heart Inhibition of tumor volumes (1024.6±378.6 vs. 749.3±234.3, P<0.001) qRT-PCR analysis showed significant up regulation of expression of CDKN1A (p21) and inhibition of expression of IGF2, NR2F2 and PLAU (uPA) Conclusion: ProstaCaid has significant anticancer activity in vivo with no signs of toxicity.

Prostate Polybotanical Formula

Breast Polybotanical Formula Contains botanicals, purified biologically active nutritional compounds and botanically enhanced medicinal mushrooms

Suppression of Proliferation & Invasive Behavior of Human Metastatic Breast Cancer Cells by Dietary Supplement BreastDefend Jiang J, Wojnowski R, Jedinak A, Sliva D; Cancer Research Laboratory, Methodist Research Institute, Indianapolis, IN, USA Department of Medicine, Indiana University. Cancer Center, School of Med., Indiana University, Indianapolis, IN, USA. Integr Cancer Ther 2011; Jun 10 (2):192 – 200.

BreastDefend Suppresses MDA-MB-231 Growth and Breast-to-Lung Metastasis in a Orthotopic Tumor Model Jiang J, Thyagarajan-Sahu A, Loganathan J, Eliaz I, Terry C, Sandusky GE, Sliva D. Oncol Rep. 2012; 28: 1139-1145. Method: Highly aggressive triple negative human breast cancer cells (MDA-MB-231) implanted into the mammary gland in mice then given the formula orally (100 mg/kg of body weight) for four weeks. Results: The formula significantly decreased tumor growth and breast to lung metastasis, and did not affect body weight or activity of liver enzymes or show any sign of toxicity in liver spleen, kidney, lung and heart tissues in mice. The cancer metastasized to the lungs in 67% of controls but only 20 percent of treated mice. The number of metastases per animal was also significantly affected by the formula. Down-regulation of primary tumor genes PLAU (urokinase plasminogen activator, uPA) and CXCR4 (C-X-C chemokine receptor-4).

Breast Polybotanical Formula

Synergistic and Additive Effects of Modified Citrus Pectin with Two Novel Polybotanical Compounds, in the Suppression of Invasive Behavior of Human Breast and Prostate Cancer Cells Jiang J, Eliaz I, and Sliva D. Cancer Research Laboratory, MRI, Indiana University Health, Indianapolis, IN, USA Amitabha Med. Clinic & Healing Center, Sebastopol, CA, USA Depart. of Med., and Indiana Univer. Cancer Center, Indiana Univer. School of Med., Indianapolis, IN, USA. Integr Cancer Ther. 2013 March 12 (2):145-52.

Prostate Polybotanical Formula * Effect on Proliferation of Human Prostate Cancer Cells PC3 growth measured using MTT assay. Cells incubated for 24 hrs with ProstaCaid (ug/ml). *P < .05

Effect on Migration of Human Prostate Cancer Cells Prostate Formula & MCP Effect on Migration of Human Prostate Cancer Cells The invasive behavior of PC3 evaluated using migration assay in the presence of (A) MCP (mg/mL) & (B) MCP (mg/mL) plus Prostacaid (10 ug/mL). B A

Breast Polybotanical Formula Effect on Proliferation of Human Breast Cancer Cells * Growth of MDA-MB-231 measured using MTT assay. MDA-MB-231 cells incubated for 24 hrs with BreastDefend (ug/mL) at the indicated concentrations. *P < .05

Breast Formula & MCP Invasive behavior of MDA-MB-231 evaluated using migration assay in the presence of (A) MCP (mg/mL) & (B) MCP (mg/mL) plus BreastDefend (20 ug/mL). B A

Modified Citrus Pectin Published Data Summary Preclinical and Clinical Research Highlights on Modified Citrus Pectin

Preclinical Studies with MCP (17 Published Articles) In vitro (Type of Cancer) In vivo (Type of Cancer) Ex vivo (Type of Cancer) 5 (Prostate*) 2 (Prostate) including 1 Lung Metastasis Study 1 (Immune; Leukemia) 2 (Breast*,**) 2 (Breast*,**) including 1 Lung Metastasis Study 1 (Angiosarcoma & Hemangiosarcom) 1 (Melanoma***) 1 (Melanoma***) including 1 Lung Metastasis Study   3 (Colon**) including 1 Lymph and 1 Liver Metastasis Study  1 (Non Cancer; Small Intestine Permeability) 2 (Non Cancer; Kidney Injury, Arteriole Fibrosis) 9 Total 10 Total 2 Total *One in vitro study reported on both prostate and breast cancer (Glinskii OV, Huxley VH, Glinsky GV. et al. Neoplasia. 2005;7(5):522-527). **One article reported two in vivo studies and one in vitro study (Nangia-Makker P, Hogan V, Honjo Y, et al. J Natl Cancer Inst. 2002 Dec 18;94(24):1854-62). ***One articles reported both an in vivo and an in vitro study (Platt D, Raz A. J Natl Cancer Inst. 1992 Mar;84(6):438-42)

Seven Clinical Studies with MCP Human Clinical Study (Number of Participants) Type of Trial Results Zhoa, et al., Altern Ther Health Med. 2008. (n = 7) Pilot trial: Lead toxicity in hospitalized children ages 5-12 Very significant decrease in blood serum levels of lead (P = .0016) and very significant increase in 24-hour urine collection (P = .0007). No adverse effects. Azémar, et al., Clinical Medicine: Oncology. 2007. (n = 49) Pilot trial: To assess the quality of life, clinical benefit and antitumoral efficacy in advanced stage solid tumors (colorectal, prostate, breast, kidney, cervix/uterine, liver, pharynx, pancreatic, melanoma, stomach, bile duct carcinoma and chondrosarcoma) 22.5% patients showed a stable disease (SD) and 12.3% patients had a SD for a period longer than 24 weeks. One patient with metastasized prostate carcinoma showed a 50% decrease in serum PSA level after 16 weeks of treatment associated with a significant increase of clinical benefit, quality of life and decrease in pain. 20.7% patients had an overall clinical benefit response associated with a stabilization or improvement of life quality. All patients tolerated the therapy well without any severe adverse events. Eliaz, et al. Forsch Komplementmed. 2007. (n = 5) Case studies: Involving lowering of lead and mercury body burden 5 case studies presented show reduction in toxic heavy metals was achieved without side effects. Gradual decrease of total body burden is believed to have played an important role in recovery and health maintenance.

Human Clinical Study (Number of Participants) Type of Trial Results Eliaz, et al. Phytother Res. 2006. (n = 8) Pilot trial: Toxic heavy metal removal in healthy population In the first 24 hours the urinary excretion of arsenic increased significantly (p < 0.05). On day 6, urinary excretion was increased significantly for cadmium (p < 0.05). Lead showed a dramatic increase in excretion (p < 0.08), and mercury approached significance on day six. Other elements analyzed including essential elements did not show any significant change in excretion. Eliaz. 228th Annual American Chemistry Meeting. August 2004. (n = 5) Pilot trial: Decrease total body burden of mercury A significant decrease in total body mercury burden was seen in all participants. The decrease was found more significant over time. The mercury burden for the group dropped 68.32% (p=0.0313). All individual completed the study, and there were no side effects reported. Guess, et al. Prostate Cancer Prostatic Dis. 2003. (n = 13) Phase II pilot trial: Lengthening of PSA doubling time in biochemical relapsed prostate cancer patients. There were no serious side effects in any patient, but three patients withdrew from the study due to abdominal cramping or diarrhea. MCP was well tolerated by the remaining 10 evaluable patients. Seven (70%) of these evaluable patients had a significantly increased doubling time of PSA compared with their times before the treatment. Strum, et al. International Conference on Diet and Prevention of Cancer. May 1999. (n = 7) Pilot Study: Lengthening of prostate-specific antigen doubling time in biochemical relapsed prostate cancer patients. PSA doubling time was lengthened by more than 30% in 4/7 (57%) of patients. One patient had a partial response, one patient had stable disease, and one patient did not respond. No adverse effects were reported.  

The role of active biomarker galectin-3 in chronic disease progression Galectin-3 Research The role of active biomarker galectin-3 in chronic disease progression

The PREVEND Study (Prevention of Renal and Vascular End-stage Disease) de Boer RA, van Veldhuisen DJ, R. T. Gansevoort RT, et al. The fibrosis marker Galectin-3 and outcome in the general population. J Intern Med. 2012 doi: 10.1111/j.1365-2796.2011.02476.x. University of Groningen, Groningen, The Netherlands

Galectin-3 Levels & Mortality from All Causes in the General Population: PREVEND Overall Average 11.9

Galectin-3 in General Population: PREVEND (number of subjects = 7,968) CHARACTERISTIC Median Gal3 TOTAL 11.9 QUINTILE-1 7.7 QUINTILE-2 9.4 QUINTILE-3 10.9 QUINTILE-4 12.6 QUINTILE-5 15.6 P DM% 3.6 2.3 3.1 4.3 6.1 0.000 MI 3.7 1.8 2.4 2.7 4.2 7.4 Hypertension 33.4 22.2 26.6 31.1 39.7 47.9 Stroke % 0.9 0.8 0.6 1.3 1.6 0.004 Systolic BP 129.2±20.2 125.0±18.1 126.6±19.0 128.6±19.6 131.3±20.6 134.9±22.5 Diastolic BP 74.0±9.7 72.1 ±9.4 73.3±9.8 74.1±9.6 75.2±9.8 75.4±9.8

Galectin-3 in General Population: PREVEND (number of subjects = 7,968) CHARACTERISTIC Median Gal3 TOTAL 11.9 QUINTILE-1 7.7 QUINTILE-2 9.4 QUINTILE-3 10.9 QUINTILE-4 12.6 QUINTILE-5 15.6 P CRP 1.29 [0.56-3.00] 0.89 [0.39-2.16] 1.04 [0.49-2.40] 1.33 [0.58-2.92] 1.53 [0.71-3.42] 1.98 [0.85-4.28] 0.000 Cholesterol 5.66±1.12 5.41±1.05 5.56±1.10 5.68±1.11 5.79±1.11 5.91±1.17 LDL 3.69±1.05 3.47±1.00 3.60±1.01 3.71±1.04 3.77±1.05 3.90±1.06 HDL 1.27 [1.03+1.56] 1.32 [1.07-1.62] 1.28 [1.04-1.57] 1.25 [1.03+1.55] 1.24 [1.03-1.53] [0.99-1.52] Triglycerides 1.16 [0.85-1.68] 1.02 [0.75-1.43] 1.11 [0.82-1.59] 1.17 [0.86-1.69] 1.23 [0.89-1.78] 1.31 [0.95-1.92]

The COACH (Coordinating Study on Outcomes of Advising and Counseling in Heart Failure) Multicenter, randomized, controlled trial conducted in The Netherlands. A prospective sub-study was designed to evaluate Galectin-3 in patients with chronic heart failure and define cut-off levels for subsequent validation studies. Galectin-3 levels were measured in 582 banked EDTA-plasma samples.

COACH Galectin-3 Sub-Study Mortality from All Causes at 1 Year in Patients with CHF

COACH Galectin-3 Sub-Study Cardiovascular Mortality or Heart Failure-Related Hospitalization at 1 Year in Patients with CHF

Galectin-3 and Cardiovascular Health DeFillipi et al, U.S. Cardiology, 7,1, 3–6 (2010) clearly indicate that circulating Galectin-3 is an important factor in fibrosis of many organs and organ systems, and that reducing circulating Galectin-3 may have an important role in remediating cardiac injury and progression to heart failure (HF). Similarly, Psarras et al, Eur. Heart J., April 26 (2011) demonstrate that reduction in Galectin-3 levels in the myocardium may reduce fibrosis in the heart and improve outlook. De Boer et al, Ann. Med., 43,1, 60–68 (2011) identify Galectin-3 as a key indicator in cardiac health. Shash et al, Eur J. Heart Fail., 12,8, 826–32 (2011) identify Galectin-3 levels as a key agent in heart failure through fibrosis. De Boer et al. Eur. J. Heart Fail., 11, 9, 811-817 (2009) link an increase in Galectin-3 expression and presence to heightened fibrosis, and heart failure. The same article links Galectin-3 to inflammation. Inflammation is the hallmark of arteriosclerosis, therefore Galectin-3 levels also contribute to coronary artery disease, peripheral artery disease, strokes, and vascular dementia.

Modified Citrus Pectin Reduces Galectin-3 Expression and Disease Severity in Experimental Acute Kidney Injury Kolatsi-Joannou M, Price KL, Winyard PJ, Long DA. Nephro-Urology Unit, UCL Institute of Child Health, London, UK PLoS ONE 2011 April 6(4): e18683.

MCP & Kidney Injury Study Background: Folic acid (FA)-induced acute kidney injury model Method: Mice given 1% MCP-supplemented water, or plain water, 1 week before FA injection Results: During initial injury phase, all FA treated mice lost weight while their kidneys enlarged secondary to renal insult Gross changes significantly lessened in MCP group MCP clearly reduced renal cell proliferation Recovery phase: MCP group showed decreased Galectin-3 expression with decreased renal fibrosis, macrophages, pro-inflammatory cytokine expression, and apoptosis

Galectin-3 Mediates Aldosterone-Induced Vascular Fibrosis Calvier L, Miana M, Reboul P, et al. Arterioscler Thromb Vasc Biol. 2013 Jan;33(1):67-75.

Aldosterone Galectin-3 MCP Vascular Fibrosis Study Background: Aldosterone is involved in arterial stiffness and heart failure. Galectin-3 plays an important role in inflammation, fibrosis, and heart failure. MCP blocks Galectin-3 Methods and Results: Rats were treated with aldosterone combined with MCP for 3 weeks. Hypertensive aldosterone-treated rats presented vascular hypertrophy, inflammation, fibrosis, and increased aortic Gal-3 expression. Those also treated with MCP treatment abolished all the above effects. Conclusion: Galectin-3 is required for inflammatory and fibrotic responses to aldosterone in vascular smooth muscle cells in vitro and in vivo. By inhibiting Galectin-3, MCP prevents vascular fibrosis.

Elevated Galectin-3: What can we do? Test for Galectin-3 levels Address general inflammation & hyperviscosity Use MCP at appropriate dosages by: Condition Galectin-3 levels Therapeutic goal

Serum Galectin-3 Testing USFDA approved blood test that measures Galectin-3 for Cardiovascular Disease

Galectin-3 Levels: Reference Ranges • Galectin-3 levels > 17.8 ng/ml are considered to be an extreme risk factor of mortality. • Ideal levels are < 14 ng/ml in the general population. • For cancer and cardiac patients, ideal levels are < 12 ng/ml. • Galectin-3 levels change in 20% of population every 3 months. Repeated testing is important.

Galectin-3 Levels: Cancer Desired levels <12.0 ng/ml Follow up on Galectin-3 levels routinely every 3-6 months

Active Cancer Levels <17.8, or not tested MCP 15g daily Levels >17.8 MCP 20 - 25g daily Put MCP Table in

Cancer Long Term Maintenance (3 years post therapy) Levels < 12.0 ng/ml MCP 5g daily Levels = 12.0 – 14.0 ng/ml MCP 10g daily Levels = 14.0 – 17.8 ng/ml MCP 15g daily Levels >17.8 ng/ml MCP 20 – 25g daily Put MCP Table in

Galectin-3 Levels: MCP Dosage (grams) Galectin-3 Test Results (ng/ml) No Known Medical Conditions Cardiovascular, Inflammation, Fibrosis, Hepatitis Active Cancer Post Cancer (3 years) <12 5 15 12.0 – 14.0 10 14.0 – 17.8 > 17.8 20 20 - 25 Not Tested 5 -10

In Conclusion: Galectin-3 and Modified Citrus Pectin Galectin-3 is a novel, active biomarker that is both a cause of multiple diseases and a diagnostic and prognostic biomarker. Modified Citrus Pectin is a proven natural Galectin-3 blocker.