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RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES , BANGALORE , KARNATAKA ANNEXURE II PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION 1 NAME OF THE CANDIDATE AND ADDRESS Dr. SNEHALIKA MORE POST GRADUATE STUDENT DEPARTMENT OF PEDODONTICS & PREVENTIVE DENTISTRY , #20, YESHWANTPUR SUBURB, TUMKUR ROAD, K.L.E SOCIETY’S INSTITUTE OF DENTAL SCIENCES, BANGALORE – 560022. 2 NAME OF THE INSTITUTION K.L.E.SOCIETY’S INSTITUTE OF DENTAL SCIENCES, BANGALORE – 22. 3 COURSE OF STUDY AND SUBJECT MASTER OF DENTAL SURGERY – PEDODONTICS AND PREVENTIVE DENTISTRY 4 DATE OF ADMISSION TO COURSE 10th JUNE 2013 5 TITLE OF THE TOPIC CORRELATION BETWEEN FOOD RETENTION AND ORAL CARBOHYDRATE CLEARANCE RATE IN CHILDREN 1 6 .BRIEF RESUME OF THE INTENDED WORK : 6.1 NEED FOR THE STUDY : There has been substantial increase in consumption of refined foods,carbonated beverages and fruit drinks which are high in fermentable carbohydrates.1 It is widely accepted that all foods containing "fermentable carbohydrates" have the potential to contribute to caries formation. Fermentable carbohydrates are present in most starches and all sugars, including those that occur naturally in foods and those added in processed foods.2 Food particles that become trapped on the surfaces of the teeth serve as reservoirs of fermentable carbohydrates, thereby permitting plaque microorganisms to continue to produce acids and prolong the cariogenic challenge to the teeth which eventually contribute to the development and progression of dental caries.3 Several studies have looked at oral clearance and retention in different individuals but the clearance pattern of children is different than those of adults. Also the food was retained in the mouth for longer periods in the children. Thus while attempting to assess the food’s cariogenicity,it is necessary to measure the length of time that the food is retained in the mouth.4 The aim of the present study is to estimate the retentive capacity and salivary carbohydrate clearance rate of various commercially available test foods popularly consumed by the child on day to day basis. 6.2 REVIEW OF LITERATURE : • Assessment of the oral clearance of various foodstuffs(glucose rinse,glucose tablet and biscuits) in children of different ages was done in comparison with adults.The study concluded that young children(3yrs-7yrs) have both slower salivary sugar clearance and larger variation in clearance among various foodstuffs than older children and adults.5 2 • A study was conducted on popular cereal based foods and fruits and total carbohydrate content of salivary expectorants following their consumption(apple, orange, banana, pasta, wholemeal bread, rice, Alpen,white bread,branflakes,cornflakes,Weetabix).The results showed that greater percentage of carbohydrate was retained following consumption of rice and breads.4 • The occurrence and distribution of mutans streptococci in child population in Africa, Europe, and North America was compared in the study.They concluded that the differences in caries experience in children of the three continents cannot be explained by the prevailing mutans streptococci species but instead should be attributed to differences in the cariogenicity of the various diets.6 • A study was done to measure sugars,starches and metabolic acids in retained food particles.The results showed that high sucrose foods(caramel,milk chochlate,chochlate-caramel bars) rapidly cleared from the teeth while high starch foods(oatmeal cookies,potato chips) retained for upto 20 minutes.7 • One study conducted to rank several reference foods according to their plaque pH response and carbohydrate retention(apple drink caramel chocolate, cookie, skimmed milk powder, snack cracker, and wheat flake) showed that apple-flavored fruit drink and chocolate were the most acidogenic foods and skimmed milk powder the least acidogenic and caramel followed by wheatflake was the most retentive and skimmed milk the least retentive.8 • Evaluation of the association between caries prevalence and intake of confectionery(mainly sugars), and sweetened baked goods(mixture of starch and sugars) in one study concluded that high intake of foods containing a mixture of starch and sugars was associated with an increased prevalence of dental caries in children.9 • The relationships among the pH response of coronal plaque-sugar and the caries status of the tooth surface and the relationship of plaque-flora and caries was examined in the study. The results indicated that increasing subject caries status is characterized by increasing plaque levels of highly-acid-tolerant, acidogenic bacteria and an increasing plaque-pH- lowering potential.10 • Another study described young children’s intakes of nonmilk extrinsic (NME) and intrinsic/milk sugars and compared the intakes of sugars by young children with and without dental caries experience.They found that neither individual nor total sugars intakes differed between the subjects with and without caries.11 3 6.3 HYPOTHESIS Research hypothesis: There is significant difference in the food retention and carbohydrate clearance among the test foods. Null hypothesis: There is no significant difference in the food retention and carbohydrate clearance between the test foods 6.4 OBJECTIVES OF THE STUDY : The study is designed to, • To estimate the total carbohydrate content of saliva collected at various time period following the consumption of test food in young children using modified anthrone reagent. • To estimate the oral retentive capacity of test foods in children. 7 . MATERIALS AND METHODS : 7.1 Sources of data : The study group includes a total of 30 children in the age group of 3-8 yrs of both the sexes visiting to the department of Pediatric and Preventive dentistry K.L.E Institute of Dental Sciences and children attending preschools. 7.2 METHODS OF COLLECTION OF DATA(INCLUDING SAMPLING PROCEDURES IF ANY) : Sample size : The study sample consists a total of 30 children in the age group 3-8 yrs. Method of collection of sample : The study sample comprise of 30 children selected using simple random sampling. 4 Inclusion criteria : Age : 3-8yrs Exclusion criteria : • • • • • • Medically compromised children. Children under chronic medications. children on antibiotics in last 3 months. children with deep carious lesions. children with early chidhood caries. Children with malocclusion. STUDY DESIGN : A survey was conducted to select the popular test foods which were consumed by the children (3-6 yrs) on day to day basis. Diet history of the child with the help of diet chart will be obtained from the parents Subjects consuming those test foods on regular basis will only be included in the study The participating 30 subjects will be subjected to following test foods in the study TEST FOODS crème sandwich cookies chochlate bars potato chips cake flavoured and sugared cornflakes Glucose biscuits Bread group For retention of the test food: Same test foods will be chosen for retention of the test food. 5 • Estimation of salivary carbohydrate The food chosen to be tested are popularly consumed items commercially available. Solid food includes crème sandwich cookies, milk chochlate bars, potato chips , cake ,flavoured cornflakes,glucose biscuits.All the foods will be possibly taken from one local store in the chain of supermarkets in Bangalore. The same solid food will be taken for carbohydrate clearance and retention study. 30 healthy young children of age 3-8 yrs will be participating in this study.No restrictions will be imposed on normal oral hygiene practice. The participants will be asked to avoid drinking/eating(except water) for 2 hr before each experimental session. Baseline Salivary samples will be collected from each subject before the administration of food.Each subject will be given 10 gram of test food.Saliva samples will be collected at 0 min after the last bolus of food swallowed.15 ml of water will be rinsed around the mouth for 15sec and expectorated. Following this,10, 20 mins saliva samples will be collected in a sterile container. Saliva samples will be collected as single expectorations (<0.5 mL) over ice.The samples will be centrifuged to clarify the saliva, and the supernatant fluid will be assayed for total carbohydrate.This sample is directly used for carbohydrate estimation using modified anthrone reagent.0.5 ml of the samples will be transferred into the test tubes containing 5ml of conc. sulfuric acid, kept in ice water to minimize the turbulence produced in reaction. Anthrone. 0.5 ml of anthrone-ethyl acetate reagent will be carefully layered over the inner side of the test tubes and gentle swirling used to hydrolyze the ethyl acetate.After proper mixing the contents will be transferred to a 1000C boiling water bath for 10 minutes, followed by 40C, for 5 minutes and to prevent condensation of moisture on the optical tubes reading,5minutes in water bath at 200C.The absorbances due to production of green furfural derivative will be measured at 610 nm on a photoelectric spectrometer.Results will be expressed in glucose equivalents calculated from the standard curve of varying glucose concentrations. One test food will be given to each subject per session and 24 hours gap will be maintained before introducing the next test food.Following procedure,oral prophylaxis & topical fluoride application will be done. • Estimation of food retention 30 same subjects will be participating in this study. Food retention will be evaluated on all of the teeth of each quadrant. The informed consent of all human subjects will be obtained and possible discomfort and risks will be fully explained to the subjects. Subjects will receive a preliminary prophylaxis for removal of 6 calculus, plaque and stains from all tooth surfaces.Each subject will be consuming the same test food as used for salivary carbohydrate estimation.Portions of food will be eaten in normal manner and the time when the last bolus of food is swallowed will be taken as 0 min.Saliva ejectors will be placed into the subjects mouths immediately and food particles remaining on all surfaces of the teeth will be recovered and placed in plastic vials.Removal of retained food will be done by scalers and curettes.The samples are then lyophilized and weighed. Subjects will be asked to brush and floss after the scaling and wait for 10 minutes before the administration of next food portion.The procedure will be repeated two times so that the retention could be determined at two minutes and five minutes after swallowing.Altogether,subjects will consume three portions of each food during one session.Following procedure,oral prophylaxis & topical fluoride application will be done. STATISTICAL TESTS THAT WILL BE USED : • The results obtained from the study will be compared and analysed statistically • Comparison in between the test food at 0,10,20 minutes will be carried out using ANOVA test • If ANOVA is significant further pairwise comparison between foods will be carried using Tukey Test(Post Hoc Test) • For the retention study,comparison between the dry weights of retained food particles of each test food will be carried using ANOVA test 7.2 DOES THE STUDY REQUIRE ANY INVESTIGATIONS OR OTHER INTERVENTION TO BE CONDUCTED ON PATIENTS OR HUMANS OR ANIMALS ? IF SO PLEASE DESCRIBE BRIEFLY . No 7.3 HAS THE ETHICAL CLEARANCE BEEN OBTAINED FROM YOUR INSTITUTION ? Yes. The ethical committee clearance certificate is enclosed 7 8. LIST OF REFERENCES : • Saha S, Jagannath GV, Shivkumar S, Pal SK. Effect of commonly consumed fresh fruit juices and commercially available fruit juices on ph of saliva at various time intervals. J Int Dent Med Res 2011;4(1):7-11. • Hegde A, Shetty R, Sequeira A R .The Acidogenicity of Various Chocolates Available in Indian Market: A Comparative Study. International Journal of Clinical Pédiatric Dentistry 2009;2(2):20-24. • Kashket S, Houte JV, Lopez LR and Stocks S. Lack of correlation between food retention on the human dentition and consumer perception of food stickiness. J Dent Res 1991;70:1314. • Pollard MA et al. Acidogenic potential and total salivary carbohydrate content of expectorants following consumption of some cereal based foods and fruits. Caries Res 1996;30:132-137. • Lingström P, Ruyven FOG, Houte JV and Kent R. The pH of Dental Plaque in its Relation to Early Enamel Caries and Dental Plaque Flora in Humans. J Dent Res 2000;79(2):770-777. • Dodds MWJ and Edga WM. The Relationship Between Plaque pH, Plaque Acid Anion Profiles, and Oral Carbohydrate Retention After Ingestion of Several 'Reference Foods' by Human Subjects. J Dent Res 1988;67(5):861-865. • Crossner CG, Hase JC, Birkhed D. Oral sugar clearance in children compared with adults. Caries Res 1991;25:201-206. • Teresa AM , Julie ME, Michelle AL, Warren JJ, Steven ML. Comparison of the intakes of sugars by young children with and without dental caries experience. JADA 2007;138:39-46. • W.H. van Palenstein Helderman, M.I.N. Matee, J.S. van der Hoeven, and F.H.M. Mikx. Cariogenicity Depends More on Diet than the Prevailing Mutans Streptococcal Species. J Dent Res 1996;75(1):535-545. • Reina Garcia-Closas. A cross-sectional study of dental caries, intake of confectionery and foods rich in starch and sugars and salivary counts of Streptococcus mutans in children in Spain. Am J Cli Nutr 1997;66:1257-63. • Kashket S, Zhang J, and Houte JV. Accumulation of Fermentable Sugars and Metabolic Acids in Food Particles that Become Entrapped on the Dentition. J Dent Res 1996;75(11):1885-1891. 8 9 Signature of Candidate 10 Remarks of the guide APPROVED 11 Name and designation of 11.1 Guide Dr. SANDHYADEVI. S. PATIL , PROF AND HEAD OF DEPARTMENT DEPARTMENT OF PEDODONTICS AND PREVENTIVE DENTISTRY, K.L.E SOCIETY’S INSTITUTE OF DENTAL SCIENCES, BANGALORE - 22 11.2 Signature 11.3 Co- Guide ( if any ) 11.4 Signature 11.5 Head Of The Department Dr. SANDHYADEVI. S. PATIL , PROF AND HEAD OF DEPARTMENT DEPARTMENT OF PEDODONTICS AND PREVENTIVE DENTISTRY,K.L.E SOCIETY’S INSTITUTE OF DENTAL SCIENCES, BANGALORE - 22 11.6 Signature 9 12 12.1 Remarks of the Chairman and Principal Dr .SRIVATSA PRINCIPAL , K.L.E SOCIETY’S INSTITUTE OF DENTAL SCIENCES, BANGALORE - 22 12.2 Signature 10