WELCOME TO 8th WORLD CONGRESS ON TOXICOLOGY AND PHARMACOLOGY

1. WELCOME TO8th WORLD CONGRESS ONTOXICOLOGY AND PHARMACOLOGY Dr. RavindraTiwari Research Cordinator UGC Research Unit Bhavan’s New Science College Hyderabad Telangana State INDIA Email- rravindra_tiwari@yahoo.com

2. “THE IMPACT OF EXTREMELY LOW FREQUENCY-ELECTROMAGNETIC FIELDS (ELF-EMFS) AND LIGHT AT NIGHT (LAN) ON ESTRADIOL (E2) LEVELS, OXIDATIVE STRESS AND DNA CONFIGURATION IN FEMALE NIGHT SHIFT WORKERS” Tiwari R1, Surender V1, Bhargava SC2, Ahuja YR3, Kavitha Varak1 1 UGC research unit, Bhavan’s new science college, Hyderabad, Telangana State, India. 2 Department of electrical and electronic engineering, SreeNidhi Institute of Science and Technology, Hyderabad, Telangana State, India. 3Genetics unit, Vasavi Medical and Research Centre, Hyderabad, Telangana State, India. “

3. INTRODUCTION An Electro Magnetic Field (EMF) is an array of waves that arise with the gathering of electric and magnetic fields, which oscillate at a specific frequency and at definite intervals with each other.

4. INTRODUCTION Phones and wireless radiofrequency devices are the most common EMF sources that are used in close proximity to the human body. Several epidemiological studies have found that long-term night working women exposed to EMFs have a higher risk of breast cancer than those who do not work at night.

5. Lack of Concentration

7. ESTRDIOL & BREAST CANCER A woman's risk for breast cancer is supposed to associated with increased levels of estrogen. Estradiol is a female sex hormone produced by the ovaries, adrenal gland and also the placenta during pregnancy. Estradiol is the most important hormone during a female’s reproductive years, and is required for reproductive and sexual function as well as having an impact on various organs and tissues.

8. BACKGROUND • Literature Review INTERNATIONAL and NATIONAL • Data inconclusive • Controversial • Lacks scientific evidence Prompts our study to rule out the conflicting reports of ELFs

9. BACK GROUND WORLD HEALTH ORGANISATION (WHO) International Commission on Non-ionizing Radiation Protection (ICNIRP) • The RF Research Agenda indicates “high priority" research for future health risk assessments. • The International Agency for Research on Cancer (IARC) classified RF field as group 2B carcinogen . • Epidemiological studies give no consistent evidence of causal relation between RF exposure and any health effect. Bio-Initiative reports are – Inconclusive.

10. OUR STRENGTH BACKGROUND • Completed two UGC Major Research Projects in the same field. 1. Study of genomic stability and hormonal configuration in human subjects exposed to extremely low frequency electromagnetic waves from different sources like mobile phones, computers and high tension wires (2005-2008). 2. Risk assessment of human subjects occupationally exposed to ELF-EMFs and light at night (LAN) (2011-2014).

11. OUR STRENGTH BACK GROUND • Corresponding Author - Published papers in the field of EMR in journals of good impact factors. Written chapters in books on effects and applications of EMR. • Referee for various research articles in journals of good impact factors and project – ICMR. • Invited speaker recently for an international conference in Beijing-China, Chicago. • One of the Papers is on the top of 20 Articles related to the same field. • Ministry of Science and Technology on behalf of society for environment and development collected data of the outcome from our major project (UGC - 2008).

12. Mobile Phones and Female Night Shift -Statistics in India India Population 1.25 billion Mobile subscribers 980+ Million Female Night Shift Workers App 11 %

13. PICO • Population:Individuals exposed to Electromagnetic Fields and Light at Night (LAN). • Intervention: To bring awareness among public, if the findings indicate a positive effect. • Comparator: a a) Women subjects occupationally exposed to EMFs and LAN:400 b) Controls (not occupationally exposed to EMFs and LAN) : 200 • Outcome:To rule out the existing debate and controversy based on our results to evaluate the mechanism of changes if any, particularly related to Estrogen levels and DNA integrity

14. OBJECTIVES • To study DNA damage in leukocytes. • Chromosomal aberrations. • Malondialdehyde [MDA] in Plasma. • Estimation of Serum Nitric Oxide. • Estimation of Estradiol levels.

15. STUDY DESIGN Qualitative (Observational): • Survey – women working in night shift in It software companies and hospitals. • Social determinants of health problems if any with the help of questionnaire. Quantitative (Experimental): • Estimations of the parameters as per the objectives • Data Interpretation. • Statistical Analysis.

16. METHODOLOGY • Subjects – a) Women subjects occupationally exposed to EMFs and LAN : 400 b) Controls (not occupationally exposed to EMFs and LAN) : 200 • Sampling - • Peripheral blood in a sterile heparinized Vacutainer tubes. • Comet assay -Single Cell Gel Electrophoresis. • Chromosomal Aberrations - MNT- Micronucleus Test. • Malondialdehyde [MDA] Assay. • Estimation of Serum Nitric Oxide. • Estradiol Levels- CLIA . C O M E T A S S A Y E S T R A D I O L L E V E L N I T R I C O X I D E MN T A S S A Y M D A A S S A Y

17. METHODOLOGY Comet Assay Sensitive test to study the degree of DNA damage devised by Singh et al (1988) with some modification after McGlyannet al (1999) RegularAgarose Low Melting Agarose Sample + LMA C O M E T A S S A Y MN T N I T R I C O X I D E MD A A S S A Y E S T R A D I O L L E V E L Lysing Alkali Treatment Electrophoresis Buffer

18. METHODOLOGY Micronucleus Test (MNT) To estimate chromosomal aberrations by slide observation method. M N T A S S A Y C O M E T A S S A Y MD A A S S A Y N I T R I C O X I D E E S T R A D I O L L E V E L GENE EXPRESSION

19. WORK DONE SO FAR MALONDIALDEHYDE [MDA] ASSAY: The oxidative stress marker, Malondialdehyde [MDA] is the stable end product of lipid peroxidation and an efficient parameter for lipid oxidative damage. Formation of MDA was assessed by quantification of Thiobarbituric Acid Reactive Species (TBARS). C O M E T A S S A Y MN T N I T R I C O X I D E E S T R A D I O L L E V E L MD A A S S A Y

20. WORK DONE SO FAR ESTIMATION OF NITRIC OXIDE: As Nitric Oxide (NO) is an unstable compound, it is rapidly converted to nitrates and nitrites in the body, hence their concentration is parallel to NO levels. Total production of NO was determined by assaying for nitrite and nitrate. Nitrate and nitrite concentrations were then estimated by the Griess method. N I T R I C O X I D E E S T R A D I O L L E V E L MN T C O M E T A S S A Y [M D A] A S S A Y

21. WORK DONE SO FAR • ESTRADIOL LEVEL: • Estrogen is a steroid hormone that is necessary for the normal development and growth of the breasts, menstrual cycle and reproductive organs essential for perpetuation of species. • Exposure to EMFs increase the level of Estrogen. • Estradiol levels were estimated by CLIA (CHEMILUMINESCENCE IMMUNO ASSAY) –[Ratcliffe et al., 1988] E S T R A D I O L L E V E L N I T R I C O X I D E [M D A] A S S A Y MN T A S S A Y C O M E T A S S A Y

22. WORK DONE SO FAR E S T R A D I O L L E V E L M D A A S S A Y N I T R I C O X I D E C O M E T A S S A Y MN T AS SAY

24. The t-value for estradiol estimation was 22.99. The p-value was <00001. The result was significant at p < .05. The alkaline comet assay indicated DNA damage ranging between 1.72µm to 9.23µm. The maximum tail length (9.923 + 0.67µm) was observed in night shift female workers.

25. CONCLUSION • The so far results suggested the induction of DNA damage and increase in frequency of micronuclei in night shift workers, probably due to genotoxic action of ELF-EMFs and LAN. • The MDA levels in plasma and No levels in Serum of exposed subjects from controls suggest a statistical significance. Thus Oxidative Stress markers could have an important implication in the comprehension of the biological stress response to ELF-EMFs and LAN. • Elevated Levels of Estradiol (E2) in Exposed compared to controls suggest to rule out the possibility of Breast Cancer.

26. CONCLUSION • This study will therefore contribute – • To Assess the Biomarkers involve in Electro Magnetic Pollution. • To envisage the Role of Estradiol in Pathophysiology of Breast Cancer.

28. Thank You! Corresponding author: 08790101015 rravindra_tiwari@yahoo.com