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RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES BANGALORE, KARNATAKA. ANNEXURE – II PROFORMA FOR REGISTRATION OF TOPIC FOR DISSERTATION 1 Name Of The Candidate And Address (In Block Letters) Dr. AYESHA N POST GRADUATE STUDENT, DEPARTMENT OF ORAL MEDICINE AND RADIOLOGY, VYDEHI INSTITUTE OF DENTAL SCIENCES AND RESEARCH CENTRE. #82 ,NALLURAHALLI ,EPIP AREA,WHITEFIELD BANGALORE-560066. 2 Name Of The Institution VYDEHI INSTITUTE OF DENTAL SCIENCES AND RESEARCH CENTRE. BANGALORE-560066. KARNATAKA. 3 Course Of Study & Subject MDS ORAL MEDICINE AND RADIOLOGY 4 Date Of Admission To Course 28-05 -2013 5 Title Of The Topic “THE INFLUENCE OF HANDHELD MOBILE PHONES ON FLOW RATE AND PROTEIN CONCENTRATION OF HUMAN PAROTID SALIVAA CROSS SECTIONAL STUDY” 6 BRIEF RESUME OF THE INTENDED WORK 6.1 Need For The Study : Mobile technology has emerged into our world rapidly and has caused many changes in our lifestyles. The handheld mobile phone operation is based on electromagnetic waves, specially radiofrequency waves and microwaves. The transmission is performed through a range of frequencies between 800 and 2200 MHz¹. The long duration and proximity of mobile phones to human body during use has given rise to concerns of possible adverse effects resulting from absorption of these emissions by the tissues adjacent to area of mobile phone handset use2. The radiation causes adverse health effects like parotid gland tumors and epithelial malignancies, auditory canal pathologies, tinnitus3. Parotid glands are the largest salivary glands, situated in front of the ear and behind the ramus of the mandible in proximity to the skin of the face. The parotid glands secrete a serous saliva and are mainly activated after stimulation resulting from smell, taste, mastication and radiofrequency radiation¹ . Saliva is mainly composed of water (99.5%) and solids (0.5%) -organic substances and inorganic substances . Normal salivary flow rate is 1ml/min. The location of parotid gland makes it especially vulnerable to changes resulting from mobile phone heat and radiation². Salivary proteins play a role in relation to protection of the teeth against caries and dental erosion. Normal saliva protein concentration is 6-8.5mg/dl. Several saliva proteins such as proline rich proteins, cystatins, MG1, lactoferrin, lysozyme and amylase take part in the formation of acquired pellicle that covers the teeth. Thus, a thick pellicle seems to be more protective against acid challenge on hard dental tissue than thin one in preventing risk of developing dental caries and erosion. A real or perceived decrease in the amount of saliva results in Xerostomia which leads to rampant caries, increased plaque formation, opportunistic fungal infections, mucositis, difficulty in swallowing and eating, malnourishment. Few studies have been conducted in western countries and very few documented in India regarding the influence of mobile phone on the salivary flow rate 1,2. Hence, the aim of this study is to evaluate the parotid salivary secretion rate and protein concentration between dominant side (more frequently used side) and non dominant side (less frequently used side) of handheld mobile phone users. 6.2 Review of Literature: Burlage F R, Pijpe J, Coopes R P, Hemles M E, Meertes H, Canninus A et al (2005) found the variability of flow rate when collecting stimulated human parotid. They estimated the accuracy and reproducibility of citric acid stimulated parotid saliva sampling and found a stronger correlation between flow rates in healthy individuals from right and left parotid glands and even more stronger relation in patients with sjogren’s syndrome4. Goldwein O, Aframian D J (2009) conducted a study to evaluate the influence of handheld mobile phones inducing physiologic changes in adjacent parotid gland. A significantly higher saliva secretion rate and lower total protein concentration was noticed in dominant side compared with non dominant side¹. Augner C, Hacker G W, Oberfeld G, Florian M, Hitzl W, Hutter J et al (2010) conducted a study to know whether exposure to radiofrequency electromagnetic fields emitted by mobile phone base stations may have effects on salivary alphaamylase, immunoglobulin A and cortisol levels in human subjects. There was significant increase of cortisol and higher concentration of alpha-amylase due to exposure. Immunoglobulin A concentration was not related to exposure5. Duan Y, Zhang H Z, Bu R F (2011) investigated the association between cellular phone use and epithelial parotid gland malignancy. They concluded that the there is a possible dose- response relationship of cellular phone use with malignancy³. Bhargava S, Motwani M B, Patni V M (2012) studied the effect of handheld mobile phone use on parotid gland salivary flow rate and volume. An significant increase in salivary flow rate along with increased blood flow rate and volume of parotid glands of the side where mobile phones are frequently placed was observed in heavy user group². Hamzany Y, Feinmesser R, Shipitzer, Mizrachi A, Hilly O, Hod R et al (2013) studied to compare salivary secretion, focusing on various salivary components and oxidative stress indices between users of mobile phones and non users. The data showed a significant increase in all oxidative stress indices studied in mobile phone users. Salivary flow was increased and total protein, albumin and amylase activity were decreased in mobile phone user . Also the use of mobile phones may cause oxidative stress and modify salivary function6. 6.3 OBJECTIVES OF THE STUDY: 1. To evaluate parotid salivary flow rate and protein concentration in healthy individuals who use handheld mobile phones. 2. To compare the salivary flow rate and protein concentration in dominant and non dominant side of handheld mobile phones. 7 MATERIALS AND METHODS: 7.1 Source of data: 7.1.1 Type of study: Cross sectional Study 7.1.2 Study area : Department of Oral Medicine & Radiology, Vydehi Institute Of Dental Sciences 7.1.3 Study population: Out patients visiting Vydehi Institute of Dental Sciences for various dental treatment 7.1.4 Duration of study: 2013-2015 7.1.5 Materials: Case History Proforma Mouth mirror, probe, tweezer Swabs Graduated tubes Test tubes Centrifuge Biuret reagent 7.2 METHOD OF COLLECTION OF DATA: (including sampling procedure if any). Sample Size: Study group- 60 healthy individuals were selected and subdivided into 2 groups Group1- 30 individuals who use handheld mobile phone for ˂ 2 hours a day Group 2- 30 individuals who use handheld mobile phone for > 2 hours a day In each group 15 males and 15 females are selected. Inclusion criteria 1. Healthy individuals aged between 19 to 35 with a history of using the handheld mobile phones for at least 3 years. Exclusion criteria: 1. Patients on any medication 2. Chronic alcohol or smoking abuse 3. Xerostomia 4. Systemic diseases 5. Past head or neck injury, trauma 6. Pregnancy 7. No preferable custom to side holding the handheld mobile phones Data collection methology: 1. 60 healthy individuals are selected and subdivided into 2 groups of which Group 1 with 30 individuals who use mobile phone for less than 2 hours and Group 2 with 30 individuals who use mobile phone for more than 2 hours. In each group 15 males and 15 females are selected, in age group of 19- 35 years with mean use of handheld mobile for 3 years are selected for the study after written consent is obtained from them. 2. A detailed case history will be recorded along with questionnaire concerning medical history, life style and habits practised. 3. The subjects will be examined during their visit to the OPD of Vydehi institute of Dental Sciences. 4. Participants will be asked to refrain from intake of any food or beverages except water, refrain from chewing gum, intake of coffee prohibited 2 hours before the saliva collection. 5. Saliva is collected between 9 AM – 12 PM for all the subjects. 6. Subjects were asked to relax for 5 minutes before salivary flow rate measurement. 7. Saliva is collected using sterile disposable containers. Prior to placement of the container, buccal mucosa is dried with gauze and stenson’s ducts are slightly squeezed to locate duct orifices. 8. Rationale for using unstimulated saliva rather than stimulated was to examine the parotid glands in their resting state ( the state in which the glands are for most part of the day, i.e. unstimulated ). 9. Saliva was collected for 3 min, samples are kept on ice 4ºc after saliva collection. Thereafter, samples should be centrifuged at 14000g for 20 min at 4˚c. 10. The supernatant is collected and protein concentration determined according to Biuret method of protein estimation7 . Laboratory procedure : The protein concentration determined according to procedure described by Biuret test. Biuret TestPrincipleThis is a general test for proteins. Cupric ion in an alkaline medium forms a violet coloured complex with peptide bond nitrogens of peptides and proteins. The reaction is so named since biuret (NH2CONHCONH2) formed by condensation of two molecules of urea when heated to 180˚c, also answers this test. The minimum requirement for a positive test is the presence of 2 peptide bonds in the molecule. Procedure4 ml of 5% NaOH in a test tube is taken and 2 drops of CuSO4 is added and mixed well. Then divided into parts. To one 1ml sample containing protein is added and to other an equal volume of water is added. Test tube containing protein gives purple colour. After the purple product is formed the protein is measured on spectrometer at 540 nm. - Result calculated as mg/dl. Statistical analysis: The values will be statistically analysed by the following tests1. Students ‘t’ test 2. Chi- Square test 7.3 Does the Study require any investigation or intervention to be conducted on patients or other human subject or animals? If so, please describe briefly. Yes, the study requires collection and evaluation of saliva from the subjects. 7.4 Has Ethical Clearance been obtained from your Institution? Yes 8. List of references: 1. Goldwein O and Aframian D J. The influence of handheld mobile phones on human parotid gland secretion. Oral Diseases 2009;16:146-150. 2. Bhargava S, Motwani M B and Patni V M. Effect of handheld mobile phone use on parotid gland salivary flow rate and volume. Oral surg Oral Med Oral Pathol Oral Radiol 2012;114(2):200-206. 3. Duan Y, Zhang HZ, Bu R F. Correlation between cellular phone use and epithelial parotid gland malignancies. Int J Oral Maxillofac Surg 2011;40 (9):966- 972. 4. Burlage F R, Pijpe J, Coppes R P et al.Variability of flow rate when collecting stimulated human parotid. Eur J Oral Sci 2005;113:356-359. 5. Augner C, Hacker G W, Oberfeld G, Florain M, Hitzl J, Pauser G. Effects of exposure to GSM mobile phone station signals on salivary Cortisol, AlphaAmylase and Immunoglobulin A. Biomed Environ Sci 2010;23(3):199-207. 6. Hamzany Y, Feinmesser R, Shpitzer T, Mizrachi A, Hilly O et al. Is human saliva an indicator of the adverse health effects of using mobile phones. Antioxid Redox Signal 2013;18(6):622- 627. 7. Navadesh M, Kumar K.S. Measuring salivary flow: challenges and opportunities. JADA 2008; 139(Suppl 2):35S-40S.