Anti-Cancer & Anti-HIV effects of ALKA V-6 Dr. C. Reed Richardson & Dr. Dhiraj Vattem TEXAS STATE UNIVERSITY San Marcos Texas
OBJECTIVES The overall objective of this research was to determine Cancer chemotherapeutic Anti-retroviral effects of ALKA-HYDROXY in cell cultures and other in vitro systems.
Anti-Cancer Effect What is the ability of ALKA V-6 to prevent mutations in the DNA? What is the ability of ALKA V-6 to prevent oxidative stress? Induce programmed cell death Activate antioxidant enzymes Increase Nitric oxide
Can ALKA V-6 kill Cancer cells?? Colon Cancer Cells (HT-29) 250000 200000 150000 100000 50000 No. of cells 100.0 Yes!! Almost completely! 80.7 % Decrease -1.1 120 100 80 60 40 20 0-20 Decrease in Live Cells (%) 0 Control T1 (1:40) T2 (1:400) T3 (1:4000) -40
Adhesion of Cancer Cells Attachment of cancer cells eventually leads to metastasis of cancer. LESS ADHESION THE BETTER!!
Colon Cancer Cells Adhesion & ALKA V- 6 250000 100 Colon Cancer Cells (HT-29) 200000 150000 100000 50000 0 no. of cells 69.0 70% decrease in adhesion! % Decrease 14.1 80 60 40 20 0-20 Decrease in attached cells (%) Control T1 (1:400) T2 (1:4000)
Mutations and Cancer Mutations Sometimes Lead to Cancer Caused by pollutants, toxins Fewer Mutant Cells the Better!!
Mutagens & ALKA V-6 Decrease in number of mutations (%) 120 100 80 60 40 20 Insertion Frameshift mutation Antimutagenic effect of ALKAVITA Missense mutation Nonsense mutation 50-100% decrease in mutations! T1 (1:40) T2 (1:400) T3 (1:4000) NaN3 =2 ug/plate Missense mutation (ALKA V-6) Deletion Frameshift mutation 0 TA98 TA100 TA102 TA1535 TA1537
Apoptosis Programmed cell death Cells normally age and die after a few divisions Defective development and progression of cancer. resistance to chemotherapy Selectively induce apoptosis in cancer cells Measured by fragmented DNA Normal Cell Cancer Cell
ALKA V-6 and Apoptosis Apoptosis increased 60 50 Control 1:5000 w/v 1:2500 w/v 1:1000 w/v 1:100 w/v Fragmented DNA (%) 40 30 20 54.1 42.1 44.8 47.6 42.8 42.1 45.9 50.1 48.8 39.5 10 0 Day 2 Day 6
Oxidative stress Altered homeostatic balance resulting from oxidant insult Free radicals in the body overwhelm antioxidant defense systems Responsible for many diseases Cancer Decrease free radicals Measured by MDA Increase antioxidant defenses Enzymes Chemicals Radicals AOX defense Imbalance
ALKA V-6 and Oxidative stress MDA (mmol/mg protein) 5 5 4 4 3 3 2 2 1 1 0-1 Dose dependent decrease in oxidative stress!! 2.8 2.8 2.8 2.8 2.4 2.4 2.2 1.6 Day 2 Day 6 Control 1:5000 w/v 1:2500 w/v 1:1000 w/v 1:100 w/v 1.3 0.9
SOD and CAT Superoxide (O 2- ) and hydrogen peroxide (H 2 O 2 ) are Toxic compounds If not removed can cause cancer Superoxide is mopped up by superoxide dismutases (SODs) Hydrogen peroxide is removed by catalase (CAT) and glutathione peroxidase. Cancer cells have high levels of metabolism that Superoxide (O 2- ) and hydrogen peroxide (H 2 O 2 ) Low levels of SOD and CAT Higher the levels of SOD and CAT the better it is!
ALKA V-6 and SOD 180 SOD (umol/mg protein) 160 140 120 100 80 60 40 20 0 Control 1:5000 w/v 1:2500 w/v 1:1000 w/v 1:100 w/v 109.1 98.7 98.5 97.9 102.2 85.5 93.2 79.6 85.5 Day 2 Day 6 Dose dependent increase in SOD!! 138.5
ALKA V-6 and CAT Catalase Activity (mu/mg protein) 200 150 100 50 0-50 -100 Control 1:5000 w/v 1:2500 w/v 1:1000 w/v 1:100 w/v 85.6 50.5 62.0 50.1 29.3 6.0 29.3 57.8 Day -21.7 2-35.7 Day 6 Significant increase in CAT!!
Glutathione Most important antioxidant Prevents conditions that can lead to diseases Cancer Cataracts human immunodeficiency virus (HIV). Detoxify various harmful chemicals Heavy metals and pollutants MORE THE BETTER!
ALKA V-6 & Glutathione Dose dependent increase in GSH!! 80 Control 1:5000 w/v 70 1:2500 w/v 1:1000 w/v GSH (mmol/mg protein) 60 50 40 30 20 10 1:100 w/v 67.6 48.2 49.8 48.2 32.5 19.5 24.5 36.8 70.0 71.3 0 Day 2 Day 6
Nitric Oxide The immune system uses nitric oxide fighting viral, bacterial and parasitic infections, Decreases proliferation of tumours Associated with learning, memory, sleeping, feeling pain, and, probably, depression. Inflammation and rheumatism. Reduce Blood Pressure Viagra MORE THE BETTER!
ALKA V-6 & Nitric Oxide Significant increase in NOx!! NOx (umol/l/mg protein) 800 700 600 500 400 300 200 100 0 Control 1:5000 w/v 1:2500 w/v 1:1000 w/v 1:100 w/v 509.7 508.0 436.7447.4 446.1 434.7 342.7 364.4 284.2 284.2 Day 2 Day 6
Anti-Retroviral effects
Anti Viral Drugs Virus evades immune system Attaches to our cells using surface sugars If this process can be inhibited infection can be prevented
Viral Envelope Different sugars Determine the shape Shape can be changed by changing sugar composition If shape changes they can no longer bind to the cell receptor They cannot hide from immune system anymore
Glucohydrolase Glycohydrolase enzymes are found in the eukaryotic host cell s Golgi apparatus Inhibition has been found to decrease the infectivity of the HIV virion Two enzymes do this: Glucosidase Glucuronidase They add sugars to the viral envelope More inhibition the better!!
ALKA V-6 & Glucosidase Inhibition of HIV envelope protein glucosylation Inhibition of glucosylation (%) 140 120 100 80 60 40 20 0 Dose dependent decrease 1-10 1-100 1-1000 1-10000
ALKA V-6 & Glucuronidase Inhibition of HIV envelope protein glucuronylation Inhibition of glucuronylation (%) 140 120 100 80 60 40 20 0 Dose dependent decrease 1-10 1-100 1-1000 1-10000
ALKA V-6 & Sugar composition Can ALKA V-6 effect the sugar composition? Looking or changes not increase/decrease Ribose Heptose Sialic acid Uronic acid
ALKA V-6 & Ribose Altered Ribose
Heptose & ALKA V-6 Altered Heptose
Sialic acid & ALKA V-6 Altered Sialic acid
ALKA V-6 & Uronic acid Altered Uronic acid