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Assessment of Oxidative Stress. Cancer Aging Inflammation Arteriosclerosis Diabetes Mucosa of the Gut Disease Hepatic Disease Ischemic Reperfusion Cardiac.

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Presentation on theme: "Assessment of Oxidative Stress. Cancer Aging Inflammation Arteriosclerosis Diabetes Mucosa of the Gut Disease Hepatic Disease Ischemic Reperfusion Cardiac."— Presentation transcript:

1 Assessment of Oxidative Stress

2 Cancer Aging Inflammation Arteriosclerosis Diabetes Mucosa of the Gut Disease Hepatic Disease Ischemic Reperfusion Cardiac Disease (cardiac infarct etc.) Respiratory Disorder Renal Disease Cranial Nerves Disease Skin Disease Multiple Organ Failure Ocular Disease (Cataract) Dr. T Yoshikawa Oxidative stress involves in various diseases

3 Aging Balance is Important Immune Disorder LOO ・ L・L・ LO ・ H2O2H2O2 1O21O2 ・O2-・O2- OH Other Antioxidant Enzymes Other Antioxidants Catalase Flavonoids Vitamin C GSH-Px Carotenoids Uric Acid SOD Vitamin E Life Style-Related Diseases Parkinson disease High blood pressure Arterial sclerosis Myocardial infarction Dementia, Cataract etc. Cancer OXIDATIVE STRESS Balance of Oxidative Stress and Anti-oxidative System

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5 Oxidative Stress Markers 8-OHdG Thymidine Glycol Acrolein Crotonaldehyde Hexanoyl-Lysine 4-hydroxy-2-hexenal 4-hydroxy-2-nonenal 7-ketocholesterol Malondialdehyde Methylglyoxal Dibromo-Tyrosine Dithyrosine DNA Oxidation Protein Oxidation Lipid Oxidation

6 Damaged DNA (8-OHdG) Blood Urine Reactive Oxygen Species Repair System Damaged Base (8-OHdG) Cell Oxidative Stress Normal DNA (dG) Normal Base (dG) DNA (dG) Anti-Oxidative System Formation of 8-OHdG

7 a) Highly specific for 8-OHdG. b) Applications: urine, serum, tissue, cultured cells, saliva… c) Assay time: 3.5 hours (New 8-OHdG Check) d) Number of samples (N=3; 18 samples / kit) e) Risk management of health and aging. f) Monitoring the effect of chemicals, UV, radiation etc… g) Assessment the effect of anti-oxidants. Patent No ; Patent No Prized ;Grand invention award (1999 Japan) ;Director-general of the science and technology agency premium (2000 Japan) Anti-8-OHdG Monoclonal Antibody (N45.1) & 8-OHdG ELISA kit Assessment of Oxidative Stress inside the body. Number of Person Urinary 8-OHdG (ng/mg CRE) Concentration of Urinary 8-OHdG (250 healthy subjects)

8 2) Cancer (8-OHdG in urine) Normal controls : / umol/mol CRE Cancer patients (before treatment): / umol/mol CRE  After initiation of therapy / umol/mol CRE. Irradiation and chemical treatment result in higher 8-OHdG formation. [Reference] : Determination of urinary 8-hydroxydeoxyguanosine by automated coupled-column high performance liquid chromatography: a powerful technique for assaying in vivo oxidative DNA damage in cancer patients. Eur J Cancer Jun;31A(6): Tagesson C, et.al. 3) Dialysis (8-OHdG in serum) Serum 8-OHdG concentration from dialysis patient is higher than that from normal controls. [Reference] : Blood 8-hydroxy-2'-deoxyguanosine is associated with erythropoietin resistance in haemodialysis patients. Nephrol Dial Transplant May;18(5): Kato A, Odamaki M, Hishida A. Diabetic patients(macro albuminuria): / ug/24h (24 h urine) Normal controls : 8.1 +/- 1.8 ug/24h [Reference] : Study of urinary 8-hydroxydeoxyguanosine as a biomarker of oxidative DNA damage in diabetic nephropathy patients. J Pharm Biomed Anal Sep 21;36(1): Xu GW, et.al. 1) Diabetes (8-OHdG in urine) Relationship between 8-OHdG and diseases (1)

9 4) Cystic fibrosis (8-OHdG in urine) Cystic fibrosis patients : / nmol/mol CRE Normal controls : / nmol/mol CRE [Reference]: Oxidative damage to DNA in patients with cystic fibrosis. Free Radic Biol Med Apr;18(4): Brown RK, et. al. 5) Alzheimer's disease (8-OHdG in tissue and cerebrospinal fluid) 1. 8-OHdG levels in the intact DNA from cerebrospinal fluid with AD (9 patients) is higher than normal controls. [Reference]: Increased DNA oxidation and decreased levels of repair products in Alzheimer's disease ventricular CSF. J Neurochem Feb;72(2): Lovell MA, Gabbita SP, Markesbery WR OHdG levels in brain tissue with AD became high. [Reference]: Oxidative alterations in Alzheimer's disease. Brain Pathol Jan;9(1): Markesbery WR, Carney JM. 6) Administration AZT to HIV Patients (8-OHdG in urine) HIV patients (Administration AZT) : 355 +/- 100 pmol/kg/day HIV patients (Avoid AZT) : 182 +/- 29 pmol/kg/day [Reference]: AZT treatment induces molecular and ultrastructural oxidative damage to muscle mitochondria. Prevention by antioxidant vitamins. J Clin Invest Jul 1;102(1):4-9. de la Asuncion JG, et.al. Relationship between 8-OHdG and diseases (2)

10 8) Periodontitis (8-OHdG in Saliva) Untreated patients with periodontitis (78 patients) : /- 0.1ng/mL Normal controls (17 patients) : /- 0.1ng/mL [Reference]: New biomarker evidence of oxidative DNA damage in whole saliva from clinically healthy and periodontally diseased individuals. J Periodontol May;73(5): M Takane, et.al. 7) Childhood Bacterial Meningitis (8-OHdG in cerebrospinal fluid) Bacterial meningitis patients’ cerebrospinal fluid 8-OHdG levels became high significantly. [Reference]: Oxidative stress in childhood meningitis: measurement of 8-hydroxy-2'-deoxyguanosine concentration in cerebrospinal fluid. Redox Rep. 2000;5(5): Tsukahara H,et.al. Relationship between 8-OHdG and diseases (3)

11 [Reference]: Increased urinary excretion of the oxidative DNA adduct, 8-hydroxydeoxyguanosine, as a possible early indicator of occupational cancer hazards in the asbestos, rubber, and azo- dye industries. Pol J Occup Med Environ Health, 1993, 6:4, Chemical Industry Workers Chemical Industry Workers’ Urinary 8-OHdG levels became High.

12 [Reference] : Levels of 8-hydroxy-2'-deoxyguanosine in DNA of white blood cells from workers highly exposed to asbestos in Germany. Mutat Res, 2000 Jul, 468:2, Oxidative stress by inhalation of asbestos

13 [Reference] : The study of DNA oxidative damage in benzene-exposed workers. Mutat Res, 1996 Oct, 370:3-4, Oxidative stress caused by benzene Benzene cause oxidative damage to DNA in peripheral blood cells.

14 [Reference] : Relationships between perceived workload, stress and oxidative DNA damage. Int Arch Occup Environ Health 2001 Mar;74(2):153-7 Influence of psychological stress 8-OHdG in peripheral blood cells from women under higher psychological stress are higher.

15 [Reference] : Detection of oxidative DNA damage in human sperm and the association with cigarette smoking. Reprod Toxicol, 1997 Sep, 11:5, Sperm DNA from smokers

16 Vegetable juice(Green Juice) reduce urinary 8-OHdG

17 Effect of lemon flavonoids to diabetic rats

18 Oxidative Stress Markers 8-OHdG Thymidine Glycol Acrolein Crotonaldehyde Hexanoyl-Lysine 4-hydroxy-2-hexenal 4-hydroxy-2-nonenal 7-ketocholesterol Malondialdehyde Methylglyoxal Dibromo-Tyrosine Dithyrosine DNA Oxidation Protein Oxidation Lipid Oxidation

19 a) Highly specific for HEL. b) Applications: urine, serum, tissue, cultured cells, saliva… c) Assay time: overnight and 2 hours d) Number of samples (N=3; 18 samples / kit) e) Risk management of health and aging. f) Monitoring the effect of chemicals, UV, radiation etc… g) Assessment the effect of anti-oxidants. Anti Hexanoyl-Lysine(HEL) Monoclonal Antibody & HEL ELISA kit Assessment of Oxidative Stress inside the body. What is HEXANOYL-LYSINE ?

20 2) Cerebral Infarction (HEL in serum) [Reference] : Effect of prophylactically administered edaravone during antegrade cerebral perfusion in a canine model of old cerebral infarction. J Thorac Cardiovasc Surg Mar;133(3):710-6, Suzuki et. al. 3) Atherosclerosis (HEL in tissue) [Reference] : Immunohistochemical detection of oxidative stress biomarkers, dityrosine and N(epsilon)-(hexanoyl)lysine, and C-reactive protein in rabbit atherosclerotic lesions. J Atheroscler Thromb Aug;15(4):185-92, Fukuchi et. al. 1) Dry eyes (HEL in cornea) [Reference] : Selenoprotein p controls oxidative stress in cornea. PLoS One Mar 29;5(3):e9911, Higuchi et. al. Relationship between HEL and diseases

21 Oxidative Stress Markers 8-OHdG Thymidine Glycol Acrolein Crotonaldehyde Hexanoyl-Lysine 4-hydroxy-2-hexenal 4-hydroxy-2-nonenal 7-ketocholesterol Malondialdehyde Methylglyoxal Dibromo-Tyrosine Dithyrosine DNA Oxidation Protein Oxidation Lipid Oxidation

22 Specifications a) Highly specific for 4-HNE. b) Clone #: HNEJ-2 c) Immunogen: 4-HNE-modified Keyhole Limpet Hemocyanin d) Subclass: Mouse IgG1(kappa) e) Form: Lyophilized Powder (containing 20 ug or 100 ug of IgG) c) Applications: Immunohistochemistry and western blotting. Anti 4- hydroxy-2-hexenal (4-HNE) Monoclonal Antibody (HNEJ-2) Assessment of Oxidative Stress inside the body. Structure of 4-hydroxy-2-nonenal

23 1) Sjögren's syndrome (4-HNE in salivary gland) [Reference]: Thioredoxin may exert a protective effect against tissue damage caused by oxidative stress in salivary glands of patients with Sjogren's syndrome, J Rheumatol Oct;34(10): , Kurimoto et. al. 3) Chronic obstructive pulmonary disease (COPD)(4-HNE in lung tissue) [Reference]: Activation of nuclear factor-kappaB in airway epithelial cells in patients with chronic obstructive pulmonary disease., Respiration. 2006;73(5):610-6, Yagi et.al. Relationship between 4-HNE and diseases 2) Active liver disease (4-HNE in liver tissue) [Reference]: M-30 and 4HNE are sequestered in different aggresomes in the same hepatocytes., Exp Mol Pathol Dec;83(3): , Amidi et. al.

24 a) Applications: Human and animal serum samples, foods and beverage samples. b) Assay time: 3 minutes c) Number of samples (N=3; 26 samples / kit) d) Monitoring the effect of chemicals, UV, radiation etc… Antioxidant Assay Antioxidant assay by Test kit for antioxidant (PAO) Principle of this assay:

25 1) Alzheimer disease (Antioxidants in plasma) [Reference]:Oxidative imbalance and cathepsin D changes as peripheral blood biomarkers of Alzheimer disease: a pilot study., FEBS Lett May 23;579(13): , Straface et. al. 2) Coronary artery disease (Antioxidants in plasma) [Reference]:Oxidative stress and its association with coronary artery disease and different atherogenic risk factors., J Intern Med Oct;256(4):308-15, Vassalle et. al. Relationship between antioxidants and diseases

26 Conclusion 1) Oxidative stress markers involves in various diseases. 2) 8-OHdG, HEL, 4-HNE are detected in various biological samples such as urine, serum, tissue, cerebrospinal fluid, saliva and cultured cells. 3) For antioxidant assay, PAO is useful. PAO is applicable for serum, foods and beverage samples. 4) Oxidative stress markers and antioxidant assay may be useful for pathological analysis, development of medicines, anti-aging research, or evaluation of functional foods.

27 Indian Distributor Address: Office#4, Green Meadows Bldg:5F, Lokhandwala Complex, Kandivali (E), Mumbai , INDIA. Contact No: Tel.: Fax: Ext: 18 Dir.: mail: Contact: Mr. Tushar Barad Applied Bioscience Consultants and Distributors

28 Thank you very much for your attention!


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