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Journal of International Dental and Medical Research Editor list of 2009

ISSN: 1309-100X

Assoc. Prof. Dr. Izzet YAVUZ Editor-in-Chief and General Director Assist. Prof. Dr. Ozkan ADIGUZEL Associate Editor and Director Prof. Dr. M. Zulkuf AKDAG, Prof. Dr. Sinerik N. AYRAPETYAN Associate Editor for Biomedical research Assoc. Prof. Dr. Refik ULKU Associate Editor for Medicine Assist. Prof. Dr. Filiz ACUN KAYA Associate Editor for Dentistry Advisory Board Betul KARGUL (TURKEY) Ferranti WONG (UNITED KINGDOM) Filiz ACUN KAYA (TURKEY) Gauri LELE (INDIA) Gulten UNLU (TURKEY) Jalen Devecioglu KAMA (TURKEY) Moschos A. PAPADOPOULOS (GREECE) Nik Noriah Nik HUSSEIN (MALAYSIA) Sabiha Zelal ULKU (TURKEY) Sadullah KAYA (TURKEY) Editorial Board Xiong-Li YANG (CHINA) Zurab KOMETIANI (GEORGIA) Gajanan Kiran KULKARNI (CANADA) Mehmet Zulkuf AKDAG (TURKEY) Yuri LIMANSKI (UKRAINE) Gulten UNLU (TURKEY) Nik Noriah Nik HUSSEIN (MALAYSIA) Rafat Ali SIDDIQUI (PAKISTAN) Jalen DEVECIOGLU KAMA (TURKEY) Ferranti WONG (UNITED KINGDOM) Betul KARGUL (TURKEY) Susumu TEREKAWA (JAPAN) Nihal HAMAMCI (TURKEY) Moschos A. PAPADOPOULOS (GREECE) Izzet YAVUZ (TURKEY) Halimah AWANG (MALAYSIA) Serdar ERDINE (TURKEY) Margaret TZAPHLIDOU (GREECE) Ozkan ADIGUZEL (TURKEY) Muhammad FAHIM (INDIA) Smaragda KAVADIA (GREECE) Sabiha Zelal ULKU (TURKEY) Abdel Fattah BADAWI (EGYPT) Sossani SIDIROPOULOU (GREECE) Emin Caner TUMEN (TURKEY) Mostaphazadeh AMROLLAH (IRAN) Medi GANIBEGOVIC (BOSNIA and HERZEGOVINA) Filiz Acun KAYA (TURKEY) Stephen D. SMITH (UNITED STATES OF AMERICA) Guvenc BASARAN (TURKEY)

Sinerik N. AYRAPETYAN (ARMENIA) Suleyman DASDAG (TURKEY) Gauri LELE (INDIA) Ali GUR (TURKEY) Refik ULKU (TURKEY) Shailesh LELE (INDIA) Murat AKKUS (TURKEY) Fadel M. ALI (EGYPT) Alpaslan TUZCU (TURKEY) Igor BELYAEV (SWEDEN) Sedat AKDENIZ (TURKEY) KING Nigel MARTYN (HONG KONG SAR, P R CHINA) Muzeyyen YILDIRIM (TURKEY) Christine Bettina STAUDT (SWITZERLAND) Senay ARIKAN (TURKEY) Karla PADILLA (MEXICO) Eylem OZDEMIR (TURKEY) Selahattin TEKES (TURKEY) Claudia DELLAVIA ( ITALY ) Feriha CAGLAYAN (TURKEY) Marco MONTANARI (ITALY) Aziz YASAN (TURKEY) Filippo BATTELLI (ITALY) Ugur KEKLIKCI (TURKEY) Ali Al-ZAAG (IRAQ) Zeki AKKUS (TURKEY) Benik HARUTUNYAN (ARMENIA) Sadullah KAYA (TURKEY) Guliz Nigar GUNCU (TURKEY) Nurten ERDAL (TURKEY)


Journal of International Dental and Medical Research

ISSN: 1309-100X

TABLE OF CONTENTS DENTISTRY Influence of Composite Restorative Materials Composition on Their Diametral Tensile Strength Values Ali A. Razooki Al- Shekhli, Isra’a A. Al- Aubi Pages 67-70 Enamel Surfaces With SEM After The Application Of Different In Vivo Stripping Methods Delal Dara KILINC, Orhan HAMAMCI Pages 71-76 Odontogenic Abscess: A Retrospective Study (549 Case Between 1998-2008) Gulten UNLU, Serkan AGACAYAK, Vedat TARI, Hilal ALAN Pages 77-80 Isolated Arcus Zygoma Fractures (9 Cases Report) Rezzan GUNER, U. Nezih YILMAZ, Ferhan YAMAN, Cigdem CETIN Pages 81-85 A Retrospective Study of Dento-Alveolar Injuries in Diyarbakir, Turkey Hilal ALAN, Gulten UNLU, Vedat TARI Pages 86-88 MEDICINE Bacterial Community And Physico-Chemical Characteristics of Muthupettai Mangrove Environment, Southeast Coast Of India Manivasagan P, Sivakumar K, Thangaradjou T, Vijayalakshmi S, Balasubramanian T Pages 89-99 The Histological Structure of Filiform And Fungiform Papillae of the Puppy Rats Which Born From the Rats Given Egf After Sialoadenectomy Ayfer AKTAS, Yusuf NERGIZ Pages 100-104 Hypolipidemic Effect of Citrullus Colocynthis Seed Powder in Alloxan Induced Diabetic Rats Jeyanthi K.A, Mary Violet Christy A. Pages 105-109 BIOMEDICAL RESEAERCH The Effect of 900 Mhz Radiofrequency (Rf) Radiation on Some Hormonal and Biochemical Parameters in Rabbits Beyza KARADEDE, Mehmet Zulkuf AKDAG, Zeki KANAY, Ayse BOZBIYIK Pages 110-115

2009 Volume 2 - Number 3


Journal of International Dental And Medical Research ISSN 1309-100X http://www.ektodermaldisplazi.com/journal.htm

RESTORATIVE MATERIALS DIAMETRAL TENSILE STRENGTH Ali A. Razooki Al-Shekhli and Isra’a A. Al- Aubi

INFLUENCE OF COMPOSITE RESTORATIVE MATERIALS COMPOSITION ON THEIR DIAMETRAL TENSILE STRENGTH VALUES Ali A. Razooki Al- Shekhli1*, Isra’a A. Al- Aubi2 1. Ali A. Razooki Al- Shekhli; B.D.S., Ph.D.; Assistant Prof., Faculty of Dentistry, Ajman University of Science & technology, UAE. 2. Isra’a A. Al- Aubi; B.D.S. , M.Sc.; Lecturer, Faculty of Dentistry, Ajman University of Science & technology, UAE.

Abstract Purpose: To evaluate and compare the diametral tensile strength (DTS) values of the newly introduced nanofilled composites with the conventional types. Methods: Eight types of dental restorative composites of A3 shade were selected in this study for (DTS) testing: Filtek Supreme XT (3M Espe), Z100 Restorative (3M Espe), Filtek P60 (3M Espe), Filtek Z250 (3M Espe), Premise (Kerr), Point 4(Kerr), Herculite classic (Kerr), and Solitaire (Heraeus-Kulzer). Eight groups of specimens (n = 10) were prepared for diametral tensile strength evaluation. Resin composite specimens were prepared by incremental (two increments) insertion of composite into a circular nickel-chromium split mold of 6 mm in inner diameter and 3 mm in height and cured for 40 seconds for each increment of composite thickness. Specimens were placed into a dark bottle containing distilled water at 37°C for 7 days. DTS tests were performed in a Universal Testing Machine (0.5 mm/min). Results: The results showed that the highest DTS values were found for the Premise composite followed by Point 4, Herculite, Solitaire, Z250, Supreme XT, P60 and Z100 which exhibited the lowest DTS values. Conclusion: The composition of light activated composites is significantly influences their DTS values. (Journal of International Dental and Medical Research 2009; 2: (3), pp. 67-70) Keywords: Composite resins; nanofilled composites; diametral tensile strength. Received date: 07.October 2009 Introduction Composite resin technology has continuously evolved since its introduction by Bowen (1963) (1) as a reinforced Bis-GMA system. A major breakthrough in composite technology was the development of photo-curable resins (2). A continued development resulted in materials with reduced particle size and increased filler loading that significantly improved the universal applicability of light-cured composite resins (3). Resin composites are widely used in dentistry and have become one of the most commonly used esthetic restorative materials because of their adequate strength, excellent esthetics, moderate cost compared with ceramics, ability to be bonded to tooth structure (4), improvements in composition, simplification of the adhesive procedures and the *Corresponding author: Assistant Prof., Ali A. Razooki Al- Shekhli Faculty of Dentistry Ajman University of Science & technology UAE. E-mail: alirazooki@yahoo.com

Volume 2 ∙ Number ∙ 3 ∙ 2009

Accept date: 02 December 2009 decline in amalgam usage due to fear of mercury toxicity (5) represent additional advantages. During the last decades, the increasing demand for esthetic dentistry have led to the development of resin composite materials for direct restorations with improved physical and mechanical properties, esthetics and durability. The latest development in the field has been the introduction of nanofilled materials by combining nanometric particles and nanoclusters in a conventional resin matrix. Nanofilled materials are believed to offer excellent wear resistance, strength and ultimate esthetics due to their exceptional polishability, polish retention and lustrous appearance (6). The essence of nanotechnology is in the creation and utilization of materials and devices at the level of atoms, molecules, and supramolecular structures, and in the exploitation of unique properties and phenomena of particles (7) with size ranging from 0.1 to 100 nanometers.The compressive and diametral strengths and the fracture resistance of the nanocomposite materials are equivalent to or higher than those of the other commercial composites tested (hybrids, microhybrids and microfill) (8). Nanofilled resin Page 67


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composites show mechanical properties at least as good as those of universal hybrids and could thus be used for the same clinical indications as well as for anterior restorations due to their high aesthetic properties (9).

RESTORATIVE MATERIALS DIAMETRAL TENSILE STRENGTH Ali A. Razooki Al-Shekhli and Isra’a A. Al- Aubi

Eighty composite specimens were prepared forming 8 experimental groups (G1-G8) (n=10) of each composite type selected respectively.

Materials and Methods Eight commercially available light-cured composite resin restorative materials, namely, Filtek Supreme XT (3M-ESPE,St. Paul, MN,USA), Z100 Restorative (3M-ESPE,St. Paul, MN,USA), Filtek P60 (3M-ESPE,St. Paul, MN,USA), Filtek Z250 (3MESPE,St. Paul, MN,USA) Premise (Kerr,orange,CA 92867,U.S.A.), Point 4 (Kerr, Italia S.p.A.), Herculite classic (Kerr,Orange CA 92867,U.S.A.), and Solitaire (Heraeus Kulzer GmbH, D-63450 Hanau, Germany) , shade A3, were tested. These materials are described in Table 1. The materials were handled according to the manufacturer’s instructions. Figure 1. Composite matrix used in this study. The fully cured composite specimens were removed from the spited mold and stored in a lightproof container with distilled water at 37°C for 1 week. Diametral tensile strength testing was performed using a Universal Testing Machine (Instron Corporation, Canton, MA) at a crosshead speed of 0.5 mm/min. Specimens were positioned vertically on the testing machine base and subjected to compressive load until failure. The diametral tensile strength (DTS) was calculated using the equation: DTS = 2L/πDh, where L is the failure load, D the diameter, and h the height of the specimen (Figure 2).

Table 1. Composition of composite resins evaluated in the present study.

The composites were placed into nickelchromium split matrixes (h = 3 mm, d = 6 mm)( Figure 1) according to ADA specification #27, item 5., with increments of 1.5 mm in thickness, cured after placement of each increment with bluephase C5 (LED) light-curing device (Ivoclar, Vivadent AG, FL9494 Schaan/Liechtenstein, Austria) for 40 seconds. The matrix was placed on a glass slab with a clear polyester strip for material’s placement. The last increment was also covered with a polyester strip and pressed with a glass slab to accommodate the Figure 2. Diagram of DTS test. R=DTS, L: load, D: diameter, h: height. composite into the matrix. Volume 2 ∙ Number ∙ 3 ∙ 2009

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Mean DTS values were expressed in MPa and data were analyzed by one-way ANOVA, followed by t- test at the 0.05 level of significance. Results Mean DTS values in MPa, standard deviation of the tested groups are presented in Table 2. The results showed that the highest DTS values were found for the Premise composite followed by Point 4, Herculite, Solitaire, Z250, Supreme XT, P60 and Z100 which exhibited the lowest DTS values (Figure 3).

Table 2. Mean DTS values for the experimental groups (MPa).

Figure 3. Mean DTS values (MPa) for the tested composites. The values obtained from the DTS testing were subjected to one way analysis of variance (ANOVA), which revealed a significant difference (P<0.05) among the experimental groups (Tables 3). Further analysis of the data was needed to examine the differences between different pairs of groups using the ( t-test analysis ) and indicated that , all pairs showed statistically significant differences ( P < 0.05 ) except pairs 2, 3, 8, 21, 22, 26, 27 and 28 showed statistically insignificant differences ( P > 0.05) Table (4). Discussion The DTS is a mechanical property used to understand the behavior of brittle materials when exposed to tensile stress commonly observed in anterior restorations. DTS is an acceptable and common test for dental composites (10-13). The results of this study demonstrated that, there were significant differences between one nanofilled composite (Premise) and all the other composites being tested in this study including the Volume 2 ∙ Number ∙ 3 ∙ 2009

RESTORATIVE MATERIALS DIAMETRAL TENSILE STRENGTH Ali A. Razooki Al-Shekhli and Isra’a A. Al- Aubi

other nanofilled composite Supreme XT (Table 4). Monomeric composition affects the degree of conversion of dental composites and quality of the restoration (14-17). In this study, the incorporation of ethoxylated Bis-GMA monomer (Table 1) could be one of the most important factors related to the relatively high average DTS values for Premise composite compared to other conventional composites or to other nanofilled composite (Supreme XT) (Figure 3). This study revealed that, composites that their organic matrix composed from only one type of monomer in a form of the ethoxylated Bis-GMA or Bis-GMA (Premise and Point4), exhibited higher DTS values compared to other composites being tested in this study that incorporating more than principal monomer and/or diluent monomers in their organic matrixes composition (Figure 3) (Table 1). In addition to the previously mentioned causes, Premise composite is the only composite in this study which showed a unique difference from the other composites being tested in this study in its incorporation of prepolymerized filler and could be considered an additional factor for its high mean DTS values (Table 1). This study also revealed that, there was no correlation found between the percentage of filler loading by volume and the DTS values (Table 1) and this finding does not coincide with the findings of Chung & Greener (14) in that, an increase of filler content in resin matrix improves DTS values. Finally, this study demonstrated an inverse relationship between the average filler particle size and DTS values of the tested composites (0.02 μm for Premise nanofilled composite) (Table 1) and Supreme (XT) nanofilled composite (clusters of 0.6 to 1.4 μm) or other conventional composites being tested in this study. Conclusions 1- The DTS of resin composites is mainly influenced by their organic monomer composition, prepolymerized filler and average filler particle size. 2- There was no correlation found between the percentage of filler loading by volume and the DTS values. References 1. Bowen RL. Properties of a silica-reinforced polymer for dental restorations. J Am Dent Assoc 1963; 66: 57-64. 2. Caughman WF, Rueggeberg FA, Curtis JW Jr. Clinical guidelines for photocuring restorative resins. J Am Dent Assoc 1995; 126: 1280-6. 3. Terry DA. Direct applications of a nanocomposite resin system: part 1- the evolution of contemporary composite materials. Pract Proced Aesthet Dent 2004; 16: 417-22. 4. Lu H, Roeder LB, Lei L, Powers JM. Effect of surface roughness on stain resistance of dental resin composites. J Esthet Restor Dent 2005;

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RESTORATIVE MATERIALS DIAMETRAL TENSILE STRENGTH Ali A. Razooki Al-Shekhli and Isra’a A. Al- Aubi

17: 102-9. 5. Baseren M. Surface roughness of nanofill and nanohybrid composite resin and ormocer-based tooth-colored restorative materials after several finishing and polishing procedures. J Biomater Appl 2004; 19:121-34. 6. Silikas N, Kavvadia K, Eliades G. Surface characterization of modern resin composites: a multitechnique approach. Am J Dent 2005; 18: 95100. 7. Zhang Y, Lim CT, Ramakrishna S, Huang ZM. Recent development of polymer nanofibers for biomedical and biotechnological applications. J Mater Sci Mater Med 2005; 16: 933-46. 8. Mitra SB, Wu D, Holmes BN. An application of nanotechnology in advanced dental materials. J Am Dent Assoc 2003; 134: 1382-90. 9. Beun S, Glorieux T, Devaux J, Vreven J, Leloup G. Characterization of nanofilled compared to universal and microfilled composites. Dent Mater 2006. In press. 10. Council On Dental Materials And Devices. New American Dental Association Specification Nº 27 for direct filling resins. J Am Dent Assoc 1977; 94(6):1191-4. 11. Aguiar FHB, Braceiro ATB, Ambrosano GMB, Lovadino JR. Hardness and diametral tensile strength of a hybrid composite resin polymerized with different modes and immersed in ethanol or distilled water media. Dent Mater 2005; 21:1098-103. 12. Soares CJ, Pizi EC, Fonseca RB, Martins LRM. A-Mechanical properties of light-cured composites polymerized with several additional post-curing methods. Oper Dent 2005; 30(3):389-94. 13. Zandinejad AA, Atai M, Pahlevan A. The effect of ceramic and porous fillers on the mechanical properties of experimental dental composites. Dent Mater 2006; 22(4):382-7. 14. Chung KH, Greener H. Correlation between degree of conversion, filler concentration and mechanical properties of posterior composite resins. J Oral Rehabil 1990; 17:487-94. 15. Chung KH. The relationship between composition and properties of posterior resin composites. J Dent Res 1990; 69(3):852-6. 16. Zandinejad AA, Atai M, Pahlevan A. The effect of ceramic and porous fillers on the mechanical properties of experimental dental composites. Dent Mater 2006; 22(4):382-7. 17. Tolosa MCCG, Paulillo LAMS, Giannini M, Santos AJS, Dias CTS. Influence of composite restorative materials and light-curing units on diametrical tensile strength. Braz Oral Res 2005; 19(2):123-6.

Table 4. t-test of the differences between different pairs of groups.

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THE EVALUATION OF ENAMEL SURFACES Delal Dara KILINC and Orhan HAMAMCI

ENAMEL SURFACES WITH SEM AFTER THE APPLICATION OF DIFFERENT IN VIVO STRIPPING METHODS Delal Dara KILINC1*, Orhan HAMAMCI2 1

Private Practice Adiyaman / TURKEY Professor Dicle University Faculty of Dentistry Department of Orthodontics Diyarbakir / TURKEY

2

Abstract Aim: Space gaining method is one of the long standing methods used in orthodontics that still preserves its currency. The space method applied as made from the approximal surfaces of teeth is among the alternative methods developed against other methods. In this study, it is aimed to evaluate the results of morphological changes formed by classical metal band stripes and air rotor stripping (ARS) over teeth between 1st and 3d months by checking Scanning Electron Microscope (SEM). Materials and Methods: 40 premolar teeth of 10 patients were applied stripping process using ARS and metal band stripes as a space method from mesial and distal surfaces. The mesial and distal surfaces of 40 premolar teeth were examined by extracting teeth in the 1st and 3rd months. Results: In terms of surface roughness statistically significant difference was found in the stripping process made by ARS and metal band stripes. (p<.05) However when it was evaluated according to the 1st and 3rd months no statistically significant difference was found. (p>.05) Conclusion: As a result of our research we were convinced that ARS system will be more beneficial in terms of patient comfort and hygiene motivation after the treatment. (Journal of International Dental and Medical Research 2009; 2: (3), pp. 71-76) Key words: Air rotor stripping, band strippes, approximal abrasion, enamel thickness. Received date: 15. July 2009 Introduction Air rotor stripping (ARS) is a technique used to remove controlled amount of proximal enamel in posterior segments to gain arch length for retracting and aligning anterior teeth. This treatment philosophy centers on overcoming the difficulties in adult extraction cases and the instability of over expansion in none-extraction cases. In addition, a posterior stripping strategy has been reported to significantly reduce treatment time when compared with premolar extraction treatment.1 The ARS can be differentiated from reproximation, slenderizing, or simply stripping in that these terms usually refer to enamel reduction procedures in the mandibulary incisor region. The main indications for the technique include reshaping the approximal contacts,2 solving Bolton discrepancy problems,3 treating mild or moderate *Corresponding author: Dr. Dara KILINÇ Private Pratice Adiyaman/TURKEY Fax: + 90 412 2488100 Phone: + 90 412 2488101/3410 Email: ddarakilinc@ gmail.com

Volume 2 ∙ Number ∙ 3 ∙ 2009

Accept date: 15 October 2009 crowding, and stabilizing the dental arch.4 Little concern over caries development is raised when the mandible incisor are slenderized as this area seldom develops caries. However, extending stripping posterior into areas that are certainly more caries prone may lead to increased caries susceptibility. With scanning electron microscopy (SEM), researches5 reported that metal strips used for gross reduction of proximal surfaces of premolars in vivo caused irreversible furrows in the enamel resulting in a significant increase plaque accumulation. A number of in vitro investigations have also examined caries formation on abraded and unabraded enamel surfaces 6-11 and have found significant increases in the rate of demineralization on abraded versus intact enamel. These findings have led others 12-14 to suggest that stripping should be done with caution, and only in patients with good oral hygiene. The aim of this study to examine the structure characteristics on the enamel surfaces that were applied stripping in the 1st and 3rd months following the two different stripping methods made by using ARS and band strippes.

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Materials and Methods Our research was planned according to total 40 teeth including the lower and upper I. Premolar teeth of 10 patients applied to our clinic for treatment suffering skeletal and dental Class I anterior crowding anomaly. The following criteria were taken into consideration in the selection of patients and teeth. The patient; 1) Should have a good mouth hygiene and skeletal and dental class I anterior crowding requiring 4 premolar extractions for treatment. 2) Should not have any systemic disease. 3) Should not have any endodontic, conservative and prosthetic restoration in all teeth. 4) Should not have any bad habits such as bruxism. 5) Should not have enamel hyperplasia and enamel thicknesses should be compatible with the research of Shillingburg et al15(table 1, 2).

Table 1. Enamel And Dentine Thickness Of Upper Teeth (M: Mesial, D: Distal, F: Facial, L: Lingual, T: Tuberculum, O: Fissur).

Table 1. Enamel And Dentin Thickness Of Upper Teeth (M: Mesial, D: Distal, F: Facial, L: Lingual, T: Tuberculum, O: Fissur). Volume 2 ∙ Number ∙ 3 ∙ 2009

THE EVALUATION OF ENAMEL SURFACES Delal Dara KILINC and Orhan HAMAMCI

It was planned to apply ARS operation to the mesial surfaces of the teeth that will be used in our study and make stripping with thin granned band stripes to the distal surfaces. After stripping the premolar teeth in the left quadrant were decided to be extracted at the end of the first month and the premolar teeth in the right quadrant were at the end off the third month. In our study, the periapical and bitewing radiographies were obtained from the teeth in the area where stripping is intended to be made.16 Enamel thicknesses and anatomic variations were evaluated in detail in the radiographies obtained. The patients with sufficient enamel thickness and who don’t have anatomic variation were selected for stripping that will be made by ARS and band stripes. The thin granned band stripes were applied through the aproximal surfaces of the teeth. The stripping obtained by band stripes was performed manually providing a parallel motion. The stripping was ended as the band stripe could move through the teeth approximal easily. Wire indicator was placed to prevent the laserations that might form in tissues and also ARS application to be a guide before the stripping as suggested by Sheridan.17-19 In ARS operation specially produced safe tipped (Stars Series) (Dentsplay Raintree Essix, Inc. Metairie, LA, USA) burs was applied water cooled to prevent the teeth from air rotor heat. The edges of these burs were designed and produced so as to prevent step formation. Stripping was made as offered by the producer firm. * Applying stripping by using safe tipped 699 L carbide burs * Initial contouring and smoothing out by safe tipped medium grid (100 µ particles of size) * Continuation of smouting out by safe tipped fine grid (30 µ particles of size) * Making the surface ending by safe tipped extra fine grid (30 µ of size) Stripping was ended in ideal distance by measuring through “space measuring gauges”, which is obtained with ARS kit. It was applied once a month with a period of 2 minutes by the help of a micro brush, providing spittle isolation accompanied by flour solution named Remin + (Raintree Essix, Inc. Metairie, LA ,USA) located inside the ARS kit. The patient was recommended not to eat anything for a hour. 1. Premolar teeth in the left quadrant were extracted at the end of the first month and the 1.premolar teeth in the right quadrant were extracted at the end of the third month. The extracted teeth were left inside 25 % gluther aldehyde solution and fixed until SEM examination. Page 72


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The mesial and distal surfaces of the samples obtained were examined in SEM device. The SEM images of the tooth, that was not applied any stripping was obtained. Randomly chosen 10 photographs were evaluated as “rough” or “less rough” by taking the SEM image that was not operated in different times as the guided. No statistically significant difference was observed among the results of the researchers. All other photos were evaluated afterwards. The data obtained were evaluated by using Chi-square test and SPSS 13.0 package program. Results

THE EVALUATION OF ENAMEL SURFACES Delal Dara KILINC and Orhan HAMAMCI

Figure 3. The SEM photographs of less rough enamel surfaces on the third month after ARS application.

The ARS applied teeth was examined as 70 % less rough, 30 % rough in the first month (Figure 1,2).

Figure 4. The SEM photographs of rough enamel surfaces on the third month after ARS application. Figure 1. The SEM photographs of less rough enamel surfaces on the first month after ARS application.

Figure 2. The SEM photographs of rough enamel surfaces on the first month after ARS application. In the third month 82.5 % less rough and 17.5 % roughness ratio was observed (Figure 3,4).

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When the ARS applied teeth operation was evaluated together as 1 and 3 months they were observed as 76.3 % less rough and 23.7 % rough. No statistically significant difference was found in the teeth applied ARS operation between 1 and 3 months (p>.05) (Table 3).

Table 3. The Evaluation of roughness after ARS Application and Stripping with Band Stripes According to Months. When the ARS applied teeth is examined as lower and upper arch, 72.5 % less roughness as seen in the lower arch and in the upper arch this ratio determined as 80 %. (p>.05) When upper and lower teeth were examined alltogether; 76.8 % less roughness was observed and ther was not statistically significant difference (p>.05) (Table 4) (Figure 5,6). Page 73


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THE EVALUATION OF ENAMEL SURFACES Delal Dara KILINC and Orhan HAMAMCI

Table 4. The Evaluation of Roughness after ARS Application and Stripping with Band Stripes in The Upper and Lower Arch. Figure 7. The SEM photographs of less rough enamel surfaces on the first month after band stripes application.

Figure 5. The SEM photographs of ARS applied enamel surfaces in the lower arch. Figure 8. The SEM photographs of rough enamel surfaces on the first month after band stripes application.

Figure 6. The SEM photographs of ARS applied enamel surfaces in the upper arch. In the band stripes applied teeth 30 % less roughness, 70% roughness was observed at the first month (Figure 7,8) and in the third month 17.5 % less roughness and 82.5% roughness was observed.(Figure 9,10) When band stripes applied teeth were evaluated together for 1 and 3 months 23.8 % less roughness and 76.2 % roughness was observed. (Table 3) When the band stripes applied teeth examined by separating as upper and lower arch in the lower arch 27.5 % less roughness was observed and this ratio was 20 % in the upper arch. When all upper and lower teeth are taken together 23.8 % less roughness was observed. No statistically significant difference was found (p>.05) (Table 4). (Figure 11,12). Volume 2 ∙ Number ∙ 3 ∙ 2009

Figure 9. The SEM photographs of less rough enamel surfaces on the third month after band stripes application.

Figure 10. The SEM photographs of less rough Page 74


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enamel surfaces on the third month after band stripes application.

Figure 11. The SEM photographs of band stripes applied enamel surfaces in the upper arch.

Figure 12. The SEM photographs of band stripes applied enamel surfaces in the lower arch. When evaluated with stripping methods in the first month 70 % less roughness was observed in ARS the first month and 30 % ratio was observed in band stripes. (p<.05) In the third month 82.5 % less roughness was observed in ARS operation and 17.5 % ratio was observed in band stripes. (p<.05) In the first and third months 76.3 % less roughness was observed in ARS operation and 23.7 % ratio was observed in band stripes (p<.05)(Table 5).

Table 5. The evaluation of roughness after ARS application and stripping with band stripes among themselves according to the months. Volume 2 ∙ Number ∙ 3 ∙ 2009

THE EVALUATION OF ENAMEL SURFACES Delal Dara KILINC and Orhan HAMAMCI

Discussion The abrasion in aproximal surfaces were observed by Begg20 in Australia Stone Age man and it was determined that there were no crowding in the regions where interproximal abrasion was observed. Today it is observed that the arch length, arc depth, with and intercanin length decreases by time21. The contraction in arch length turns into crowding and anomalies by time22. Today the procedures depending on slenderizing the enamel thickness in orthodontics practice is used more efficiently for the solution of crowding from minor to mild level among clinicians. 23 In the border line cases events that extraction decision can not be taken to gain the needed space for treatment without expansion, stripping 0.25 mm from the aproximal surfaces of the teeth was recommended24. In the border line cases that were decided to be treated by ARS 0.50 mm stripping was recommended from the approximal surfaces of the teeth.17 Straud et.al 25 measured the thickness of enamel and dentine and reported that the enamel thickness in the distal surfaces of the teeth was more and space gainning of 8 mm could be solved by abrasion of the enamel of the teeth at a ratio of 50 % and by abrasion of the molar teeth. Morphological changes in enamel as a result of stripping procedures are inevitable. As in our study the researchers3, 4 evaluated the examination of teeth surface as qualitative studies depending on observation in terms of morphology. These morphological changes carry importance in terms of the future life comfort of the patient. Because the increase in the amount of roughness in the enamel increases colonization 26-28 and forms microbial colonization approximately 25 times compared to a smooth surface27. It is indicated that this colonization increases periodontal problems and decay incidence.26-28 But there is also studies that don’t contain any problem as a result of the stripping in the follow-up containing 2-5 years.29 Jarjoura et al30 called the patients for controlling in once in six months who have ARS applied teeth and had no operation and didn’t observe any problems. Research reported that the secondary enamel formed after abrading on enamel surface was more resistant against decay29. In the research of the Zacrisson et al.31 no decay formation in teeth was seen after strippind in a period of 10 years follow-up. We also didn’t deserve areas inclined to decay or periodontal problems over teeth as a result of 3 monthly controls. We think that the bacterial colonization will be prevented and integrity will continue by a good awareness raising, flour application and motivation. Page 75


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As a result of strpping abrasive troughs were seen in teeth. In our study variable ratios of wells and troughs were observed. Radlanski5 indicated in SEM study that the traces occurring after abrading with a thick granned drill on enamel surface could be decreased by a thinner granned drill but could not be removed completely. Arman et al.28 in their SEM surface examination reported that cavity and spurs on the surface of the enamel can be observed even in a good polishing operation. Moreover abrading traces can be followed in a ten year followup.32 Radlanski et al.5 observed irreversible changes in stripping made by band stripes and indicated the inclination to the formation of plaque accumulation and dental caries. We believe that the difference between the stripping surfaces made by ARS and band stripes arises from using polishing techniques in ARS operation as mentioned by the researchers. One of the complications that may form in stripping operation was reported as sensitivity. In our study, no sensitivity was observed in the information obtained from patients till the teeth extraction period. We believe that as recommended by Rossouw and Tortorella33 enamel thickness was determined by obtaining bitewing radiographies and so possible complications may be prevented and this situation arises from the lack of tubular and nerve ending inside the enamel tissue in case of the ARS operation is made in disciplined approach. Conclusion As a result we believe that while in all teeth applied ARS and band stripes areas causing plaque accumulation and retention are observed; the ARS applied surfaces can give more positive results in terms of self-healing. References 1.

Winter WW. The artistry of tooth reshaping for beauty and to gain space. Lancer J 1990; 3:1-4. 2. P.E. Rossouw and A. Tortorella, Enamel reduction procedures in orthodontic treatment, J Can Dent Assoc 2003; 69:378–83. 3. D.L. Tuverson, Anterior interocclusal relations. Part I, Am J Orthod 1980;78:361–70. 4. H. Peck and S. Peck, An index for assessing tooth shape deviations as applied to the mandibular incisors, Am J Orthod 1972;61:384–401. 5. Radlanski RJ, Ralf R, Jager A, Zimmer B. Plaque accumulations caused by interdental stripping. Am J Orthod Dentofac Orthop 1988;94:416-20. 6. Kapur KK, Ficher E, Manly RS. Effects of surface alteration on the permeability of enamel to a lactate buffer. J Dent Res 1961;40:1174-82. 7. Wickwire NA. Effect of orthodontic reduction procedures on the permeability of enamel. (Thesis) Dallas, Texas: Baylor University,1964 8. Rodgers GA, Wagner MJ. Protection of stripped enamel surface with topical flouride applications. Am J Orthod 1969;56:551-9. 9. Sullivan HR. The solubity of enamel surfaces. J Dent Res 1954;33:504-10. 10. Brudevold F. A study of phosphate solubility of the human enamel

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THE EVALUATION OF ENAMEL SURFACES Delal Dara KILINC and Orhan HAMAMCI

surface J Dent Res 1948;27:320-9. 11. Isaac S, Brudevold F, Smith FA, Gardner DW. Solubilityrate and natural fluoride content of surface and subsurface enamel. J Dent Res 1958;37:254-63. 12. Hudson AL. A study of the effects of mesiodistal reduction of mandibulary anterior teeth. Am J Orthod 1956;42:615-24. 13. Tuverson DL. Anterior interocclusal relation. Am J Orthod 1980;78:361-70. 14. Paskow H. Self aligment following interproximal stripping. Am J Orthod 1970;58:240-9. 15. Shillingburg HT Jr, Jacobi R, Brackett SE. Fundamentals of Tooth Preparations for Cast Metal and Porcelain Restorations. Carol Stream, Ill: Quintessence Publishing Co; 1987. 16. Twessne DA, Firestone AR, Heaven JT, Feagin FF, Jacobson A. Air rotor stripping and enamel demineralization Am J Orthod Dentofac Orthop 1994;105:142-52. 17. Sheredian JJ. The physiologic rationale for air rotor stripping J Clinc Orthod 1997;31:609-12. 18. Sheredian JJ. Air rotor stripping J Clin Orthod 1985;19:43-59. 19. Sheredian JJ. Air rotor stripping update. J Clin Orthod 1987;21:781-8. 20. Begg PR. Stone age man’s dentition Am J Orthod 1954;40:298312. 21. Richardson ME. Late lover arch crowding: Facial growth or forward drift? Eur J Orthod 1979;4:219-25. 22. Rossouw EP, Preston CB, Lombard CJ, Truter JW. A longitudinal evaluation of the anterior border of the dentition. Am J Orthod 1993;104:146-52. 23. Peck H, Peck S. An index for assessing tooth shape deviations as applied to the mandubulary incisor. Am J Orthod 1972;61:384401. 24. Proffit WR, Fields Jr HW. Contemporary Orthodontics Mosby Edition 2nd. Edition 2000 pp:82,83,105,575,576. 25. Straud JL, English J, Buschang PH. Enamel thickness of the posterior dentition: It’s implications for nonextraction treatment Angle Orthod 1988;68:141-6. 26. Leknes KN. The influence of anatomic and iatrogenic root surface characteristics on bacterial colonization and periodontal destruction: a review J Periodontol 1997;68:507-16. 27. Quıryvev M, Bollen CM, the influence of surface roughness and surface free energy on supra and subgingival plaque formation in man. A review of literature J Clin Periodontol 1995;22:1-14. 28. Arman A, Cehreli BS, Özel E, Arhun N, Çetinşahin A, Soyman M. Qualitive and Quantitive evaluation of enamel after various stripping methods. Am J Orthop Dentofac Orthop 2006;130: 131.e7-131.e14. 29. Crain G, Sheridan JJ. Susceptibility to caries and periodontal disease after posterior air rotor stripping. J Clin Orthod 1990;24:84-5. 30. Jarjoura K, Gagnon G, Nieberg L. Caries risk after interproximal enamel reduction, Am J Orthod Dentofac Orthop 2006;130:26-30. 31. Zachrisson BU, NyØygaard L, Mobarak K. Dental health assessed more than 10 years after interproximal enamel reduction of mandibular anterior teeth. Am J Orthod Dentofac Orthop 2007;131:162-9. 32. Thordarson A, Zachrisson BU, Mjör IA. Remodelling of canines to the shape of lateral incisors by grinding: a long term clinical and radiographic evaluation. Am J Orthod Dentofac Orthop 1991;100:123-32. 33. Rossouw EP, Tortorella A. Enamel reduction procedures in orthodontic treatment J Can Denr Assoc. 2003;69:378-83.

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ODONTOGENIC ABSCESS Gulten UNLU, et al

ODONTOGENIC ABSCESS: A RETROSPECTIVE STUDY (549 CASE BETWEEN 1998-2008)** Gulten UNLU1*, Serkan AGACAYAK2, Vedat TARI2, Hilal ALAN3 1. Professor, DDS PhD, Dicle University, Faculty of Dentistry Departmant of Maxilofacial Surgery Diyarbakir / TURKEY 2. Research Assistant, MsC, Dicle University, Faculty of Dentistry Departmant of Maxilofacial Surgery Diyarbakir / TURKEY 3. Research Assistant, MsC, PhD, Dicle University, Faculty of Dentistry Departmant of Maxilofacial Surgery Diyarbakir / TURKEY

Abstract Purpose: The purpose of this study was to prospectively evaluate a series of patients with severe odontogenic abscess. Patients and Methods: In this study, we retrospectively evaluated patients with odontogenic abscess treated at the Oral and Maxillofacial Department of Dicle University during a ten-year period between 1998 and 2008. In this study 549 patients were included. Among the patients, 324 (59.0%) were males and 225 (41.0%) were females. Standardized data collection included surgical and medical treatment, complications and clinical features information. Appropriate descriptive statistics were computed. Results: In 10 years (from 1998 to 2008) 549 patients were treated. Among the patients, 324 (59.0%) were males and 225 (41.0%) were females. Patients were treated by 67% (368) extraoral drainage and 33% (181) intraoral drainage. The most common abscess formation sites were mandibular posterior (74.8%-441 case), maxillar posterior (9.6%-53 case); maxillar anterior (8.9%-49 case) and mandibular anterior (6.5%-36 case) regions. Conclusions: This study indicated that severe odontogenic abscess can be life threatening. Intra oral and extra oral drainage may be useful for successful treatment of extensive odontogenic infection, which can be life threatening when medical therapy is ineffective. (Journal of International Dental and Medical Research 2009; 2: (3), pp. 77-80) Keywords: Odontogenic abscess. Received date: 19. August 2009 Introduction Most published cases of series of severe odontogenic abscess are retrospective (1, 2). As such, they were subjected to errors due to missing data, misclassification or misinterpretation of clinical records, and inconsistent treatment methods. Therefore, we designed a retrospective and descriptive study of 549 consecutive cases of severe odontogenic abscess, defined as those

*Corresponding author: Prof. Dr. Gulten UNLU, DDS Phd Dicle University, Faculty Of Dentistry, Department of Maxilofacial Surgery, 21280 Diyarbakir, TURKEY. Tel: +90 412 248 81 01-3486 Fax: +90 412 248 81 00 E-mail: gunlu@hotmail.com **This study was presented at VIII. Congress of Association of Oral Surgery (2008 Bodrum / TURKEY)

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Accept date: 25 October 2009 warranting hospital admission. Standardized data were collected from each case, and uniform treatment methods were used (3-5). The specific aims of this study were: 1) to accumulate prospective and descriptive data to characterize severe odontogenic abscess, and 2) to determine the drainage for treatment of severe odontogenic abscess. Our hypothesis was that, surgical incision and drainage combined with prompt medical treatment of all affected anatomic spaces, would result in improvement in swelling, fever, and white blood cell count (WBC) by 48 hours after surgery(6-8). Patients and Methods STUDY DESIGN/SAMPLE In this study, we used a retrospective case series design, in which all consecutive patients with odontogenic abscess, severe enough to justify hospital admission, were treated with incision and drainage and medical treatment (unless allergic) of all affected anatomic deep fascial spaces as soon Page 77


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1) development of an allergic or toxic reaction to the antibiotic; 2) development of necrotizing fasciitis, in which case broad-spectrum antibiotic therapy was indicated; or 3) no improvement of temperature, WBC, and swelling after 48 hours of continuous medical therapy with the same antibiotic, plus a postoperative CT scan demonstrating adequate surgical drainage of all anatomic deep fascial spaces affected by cellulitis or abscess (10, 13). If inadequate surgical drainage was shown on postoperative CT scan, then the operation was repeated, with appropriate drainage of all spaces affected by cellulitis or abscess (Figure 1). 60

55

50

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35

34

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26 23

20 17

23

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p=0.001 10

13

12

13 10

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7 3

0

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20

20

20

20

20

20

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19

19

07

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02

01

00

99

98

Treatment Methods

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as possible during the end of treatment. The subjects enrolled in this study were presented for care between January 1998 and January 2008 at the clinic of Oral and Maxillofacial Surgery, Faculty of Dentistry, University of Dicle in Diyarbakir. A total of 549 subjects were enrolled in this study based on the following criteria: severe odontogenic abscess (as determined by an attending oral and maxillofacial surgeon) and hospital entrance. Informed consent was obtained by using forms and procedures developed for this institutional review board–approved study. The criteria for hospital entrance were: odontogenic abscess causing swelling in one or more of the deep fascial spaces of the head and neck, impending threat to the airway or vital structures, fever greater than 101°F, need for inpatient control of a concomitant systemic disease. Potential subjects were excluded from this study according to the following criteria: pregnancy, nonodontogenic cause (e.g. trauma-related or upper respiratory infection), and refusal of consent. Previously published nomenclature and descriptions of the deep fascial spaces were used for the purposes of this study (3, 4).

ODONTOGENIC ABSCESS Gulten UNLU, et al

Year

All patients were subjected to the same treatment protocol. The patient was prepared for surgery as soon as possible after hospital admission. Appropriate preoperative medical workup was performed, including history and physical examination, complete blood cell count, urinalysis, appropriate imaging studies, and medical consultation when necessary. Preoperative imaging methods included periapical and panoramic dental x-rays, as well as preoperative computed tomography (CT) scanning in selected cases. After establishment of a secure airway, the skin and mucosa were prepared with antiseptic solution. Drainage was performed for all anatomic fascial spaces that were involved by either cellulitis or abscess. Specimens for culture and sensitivity testing were harvested by either aspiration or by swab sampling of open surgical wounds (6-11). All spaces that opened were copiously irrigated and maintained using latex Penrose or Jackson-Pratt type drains (12). Postoperative CT scanning was performed when indicated based on the patient’s progress and response to treatment. All patients received medical treatment unless they gave a history of antibiotic allergy or presented with signs and symptoms of necrotizing fasciitis. Medical therapeutic failure was defined as: Volume 2 ∙ Number ∙ 3 ∙ 2009

Figure 1. Distribution of the cases according to different parameter by the years. DATA COLLECTION The demographic variables recorded were gender. Preadmission variables were: smoking, drug allergies, preadmission antibiotic therapy, and the presence of immunocompromising diseases (such as diabetes, human immunodeficiency virus [HIV] seropositivity (14), use of immunosuppressive medications, severe kidney disease, and cancer chemotherapy within the previous year). The timerelated variables included the number of preoperative days of pain, preoperative days of swelling, length of stay, operating room time, the time between admission and surgery, and season of the year. Preoperative clinical variables included causative teeth, number of teeth involved, dental diagnosis (such as caries, periodontal disease, or pericoronitis), dyspnea, dysphagia, trismus (maximum interincisal opening _20 mm), WBC, and admission core temperature. For purposes of statistical analysis, certain variables were grouped together. For example, upper teeth were grouped into categories of anterior maxillar, posterior maxillar. Anamnestic data were obtained from the Page 78


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subjects in a standardized fashion, limited to the current episode of infection, and verified by the attending surgeon. The recorded treatment variables included the anatomic spaces drained intraoral or extraoral drainage (18). The odontogenic abscess was categorized to be originated from mandibular posterior, maxillar posterior, maxillar anterior and mandibular anterior regions. Statistical evaluation was made with SPSS (Statistical Package for Social Sciences) 11,5 computer program. Chi square (or Fisher’s exact) test used for the analyses of categorical variable and Spearman’s correlation test was used for correlation between groups (Table 2).

ODONTOGENIC ABSCESS Gulten UNLU, et al

of infection arising from maxillary and mandibular teeth (7, 8) Conclusions The results of this study indicated that severe odontogenic abscess can be life threatening. Intraoral and extraoral drainage may be useful to successful treatment extensive odontogenic infection, which can be life threatening when medical therapy is ineffective. Socioeconomic factors, particularly ignorance, illiteracy, and poverty, are important contributory factors towards the high incidence of odontogenic abscess in developing countries.

Results A total of 549 subjects (324 males, 225 females) were enrolled in this study. Patients were treated by 67% (368) extraoral drainage and 33% (181) intraoral drainage. The most common abscess formation sites were mandibular posterior (74.8%-441 case), maxillar posterior (9.6%-53 case); maxillar anterior (8.9%-49 case) and mandibular anterior (6.5%-36 case) regions (Table 1). The design of this study did not include a prospective evaluation of the utility of computed tomography (CT), periapical and panoramic dental x-rays in the pre-and postoperative evaluation of severe odontogenic abscess. This may have led to misclassification of the anatomic location of infection in some cases, which is a limitation of this study. The accuracy of abscess detection in head and neck infections is improved by the combination of clinical examination and contrastenhanced CT (5, 6). During the course of the study, however, we observed several advantages of CT. In addition, postoperative CT was very useful in identifying correct drain placement as well as undrained loculations of pus or extension of infection during treatment. In descending necrotizing mediastinitis, Freeman et al (9) reported a reduced mortality of 0 in 10 cases, using a protocol of open thoracotomy for direct mediastinal drainage and postoperative CT taken every 48 to 72 hours in patients with the lack of clear improvement after surgery. They reported using ranges of 3 to 15 CT scans per case and 4 to 8 operations per case. Laparotomy for extension of infection into the abdominal cavity was necessary in 30% of cases, and CT was most useful in identifying the need for laparotomy (7) Recently, CT has been used to trace the anatomic pathways of the spread Volume 2 ∙ Number ∙ 3 ∙ 2009

Table 1. Distribution of the cases according to different parameter by the years. Symmetric Measures

Interval by Interval Ordinal by Ordinal N of Valid Cases

Pearson's R Spearman Correlation

Value .665 .640 40

Asymp. a Std. Error .059 .121

b

Approx. T 5.488 5.133

Approx. Sig. .000c .000c

a. Not assuming the null hypothesis. b. Using the asymptotic standard error assuming the null hypothesis. c. Based on normal approximation.

Table 2. Year, according to the different distribution within Localization are highly correlated (r=0.640, p<0.0001). References 1. 2.

3.

4.

Huang TT, Liu TC, Chen PR, et al: Deep neck infection: Analysis of 185 cases. Head Neck 2004; 26:854. Chen MK, Wen YS, Chang CC, et al: Predisposing factors of lifethreatening deep neck infection: Logistic regression analysis of 214 cases. J Otol 1998; 27:141. Flynn TR, Liu TC, Shanti RM, Levy M, et al: Severe Odontogenic Infections, Part One: Prospective Report. J Oral Maxillofac Surg 2006; 64:1093. Flynn TR, Liu TC, Shanti RM, Levy M, et al: Severe Odontogenic Infections, Part Two: Prospective Report. J Oral Maxillofac Surg

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5.

6.

7.

8.

9.

10. 11. 12.

13.

14.

ODONTOGENIC ABSCESS Gulten UNLU, et al

2006;64:1104. Flynn TR: Anatomy and surgery of deep fascial space infections, in Kelly JJ: Oral and Maxillofacial Surgery Knowledge Update 1994, American Assocation of Oral and Maxilofacial Surgeons, Rosemont, IL 1994; 79-107. Flynn TR: Anatomy of oral and maxillofacial infections, in Topazian RG, Goldberg MH, Hupp JR (eds): Oral and Maxillofacial Infections. Ed 4. Philadelphia, PA, Saunders, 2002; 188-213. Obayashi N, Ariji Y, Goto M, et al: Spread of odontogenic infection originating in the maxillary teeth: Computerized tomographic assessment. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2004; 98:223. Yonetsu K, Izumi M, Nakamura T: Deep facial infections of odontogenic origin: CT assessment of pathways of space involvement. AJNR Am J Neuroradiol 2002; 23:468. Lewis MAO, MacFarlane TW, McGowan DA: Quantitative bacteriology of acute dentoalveolar abscesses. J Med Microbiol 1986; 21:101. Flynn TR, Halpern LR: Antibiotic selection in head and neck infections. Oral Maxillofac Surg Clin N Am 2003; 15:17. Flynn TR, Stokes LN, Lee AM, et al: Molecular microbiology of orofacial infections. J Oral Maxillofac Surg 2002; 60:72. Flynn TR , Hoekstra W, Lawrence FR: The use of drains in oral and maxillofacial surgery: A review and a new approach: J Oral and Maxillofacial Surgery 1983; 41: 508-511. Freeman RK, Valliere E, Verrier ED, et al: Descending necrotizing mediastinitis: An analysis of the effects of serial surgical debridement on patient mortality. J Thorac Cardiovasc Surg 2000; 119:260. Carey JW, Dodson TB: Hospital course of HIV-positive patients with odontogenic infections. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2001; 91:23.

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ARCUS ZYGOMA FRACTURES Rezzan TANRIKULU et al.

ISOLATED ARCUS ZYGOMA FRACTURES (9 CASES REPORT)* Rezzan GUNER1, U. Nezih YILMAZ2, Ferhan YAMAN3**, Cigdem CETIN4 1. Assoc. Prof. Dr. Department of Oral and Maxillofacial Surgery, Faculty of Dentistry, Dicle University, Diyarbakir / TURKEY 2. PhD. DDs. Department of Oral and Maxillofacial Surgery, Faculty of Dentistry, Dicle University, Diyarbakir / TURKEY 3. Assist. Prof. Dr. Department of Oral and Maxillofacial Surgery, Faculty of Dentistry, Dicle University, Diyarbakir / TURKEY 4. PhD. DDs. Oral and Maxillofacial Surgery, Private Practice, Diyarbakir / TURKEY

Abstract Arcus zygomaticus is one of the weakest parts in the facial bone that can be easily affected and fractured by traumas. The typical shape of the fracture is midline depression and separation from zygomatic and temporal bone. Sometimes by the whole arcus fracture can be occur. The arcus fractures can be seen the component of the zygomatic complex fracture and also may happen as an isolated damage even if occurs minimal trauma to the lateral face parts. Most of the cases, as a result of fracture the arcus zygomaticus force the coronoid process and result in limited mouth opening and trismus. If these kinds of symptom will be seen the treatment is necessary. Conventionally, Keen, Gilles, Hook traction and open reduction treatment method and also new treatment modality can be use. For satisfied functional and aesthetically clinical results, meticulous approach is necessary. Our study’s aim is to present 9 isolated arcus zygoma fracture cases a point of clinical findings, treatment methods, and result. In addition, classification and treatment methods will be discussed in the light of literature reviews. In our study group we have 1 female and 8 male patient. Etiological factors are violence, falling, and sport accidents respectively 6 cases, 1 and 2 cases. Main compliment was limiting of the mouth opening in all patients. Keen’s method (Intraoral approach) and hook tractions method were preferred as a choice of treatment modalities. Only for one case, which is delayed arcus zygomaticus fracture, temporal approach was performed. After the operation along to 5-7 day, we performed intermaxiller fixation all the patients. After operation maximal mouth opening and functional rehabilitation was obtained. (Journal of International Dental and Medical Research 2009; 2: (3), pp. 81-85) Key words: Arcus Zygoma, Midface fracture. Received date: 20. September 2009 Introduction The zygomatic arch, contradistinction to the zygoma, is a relatively weak part of the facial bone. Fracture of the zygomatic arch and does occur

**Corresponding author: Assist. Prof. Dr. Ferhan YAMAN Dicle University, Faculty Of Dentistry, Department of Maxilofacial Surgery, 21280 Diyarbakir, TURKEY. Tel: +90 412 248 81 01-3496 Fax: +90 412 248 81 00 E-mail: dtferhan@hotmail.com *This paper was presented in Oral and Maxillofacial Surgery Society 1st International Congress 2007, Antalya / TURKEY

Volume 2 ∙ Number ∙ 3 ∙ 2009

Accept date: 09 Nowember 2009 zygomatic and temporal suture, as well as along the full length of the arch (1). Usually those M shaped fractures mentioned in text books and having 3 fractures line2. Fracture of the zygomatic arch often occurs as a part of tripod fracture of the zygoma and of the Le Fort III- type maxillary fractures (2). Isolated zygomatic arch fractures compromise about 10 % of all zygoma fractures (3). A displaced zygomatic arch fractures can be clinically diagnosed by observation of in the region by pain, prevention coronoid from moving forward as in jaw opening and by trismus caused by trauma to the temporalis muscle1. Indeed , trismus has been reported to occur in approximately half of isolated zygomatic arch fracture. The zygomatic arch fractures should treat to restore function and aesthetic (1). Facial fractures have recently been classified in fine detail according to computed Page 81


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tomographic findings. Nevertheless, there exists no classification of the zygomatic arch fracture, which has a physiognomically important place, to provide guidance for treatment (2,3). Because of these reasons new classification systems are improved: The classification according to dislocation of fracture is summarized a below (3). 1 Type I : No displacement Type II : Displacement with bone contact at all fracture lines Type III : Displacement without bone contact at 1 fracture line Type IV : Displacement without bone contact at 2 fracture lines Type V : Comminution or displacement without bone contact at 3 or more fracture lines Numerous techniques have been reported for the reduction of zygomatic arch fractures, such as closed reduction by the Gilles method or an intraoral approach percutaneous approach (hook traction) and open reduction (4). The patient should be given a soft diet for 810 days postoperatively to limit function of the masseter muscle which pulls downward to zygoma(4). Fixation by the intraosseous wiring rarely is required. The incision for open reduction lies above the zygomatic arch; care must be taken avoid the branches of the facial nerve .When exposed the soft tissue not be separated excessively lest the bone fragment loose their blood supply(4).

ARCUS ZYGOMA FRACTURES Rezzan TANRIKULU et al.

Figure 1. Lateral depression view as a preoperative.

Figure 2. Postoperative view of patient. Case Reports 9 patient, were included in the study. One of them was female and the others were male. Mean age of all patient was 33 year old (Table-1). The most common causes of fractures were violence and the fallows were sport accident and falls. Limited mouth opening was observed as symptom all patients. As a result of fracture depression of lateral depression was detected in all patient. (Fig- 1,2). The others symptom which were associated with fractures were pain and edema. On the radiological evaluation of all patients we used axial CT for 5 patients and conventional radiography (submento-vertical graph) for 4 patients. Except for patients with multi-fragmented arcus zygoma, “M” shaped depression was observed in all patients (Fig-3,5). 3 patients were treated by hook traction (Fig3,4), 5 patients were treated by Keen modality (intraoral approach) (Fig-5,6) and also one patient who was applied to our clinic on the 12th day of fracture was treated by Gillies's method (Fig- 7,8). Volume 2 ∙ Number ∙ 3 ∙ 2009

Figure 3. Multi-fragmented arcus zygoma. To prevent to relapse intermaxillary fixation (IMF) was performed to all patients for 5-7 days in the post operation period. Finally we detected that on clinical observations normal function and esthetic results had been obtained far all patients. Complete anatomic reduction was ensured on treated with Keen and Gillies’s approach. (Fig5,6,7,8) Page 82


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ARCUS ZYGOMA FRACTURES Rezzan TANRIKULU et al.

Figure 4. Overcorrection was seen on 2 of 3 Figure 7. patients treated with hook traction modality(Fig-3, 4).

Figure 8. Figure 5. Complete anatomic reduction was ensured on treated with Keen and Gillies’s approach (Fig. 5-8).

Figure 6. Volume 2 ∙ Number ∙ 3 ∙ 2009

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symptoms, diagnosis and treatment modalities on our patients (* Submentovertical). Discussion Arcus zygoma has a thin structure and also is effected easily by traumas. Therefore among facial fractures zygomatic arch fractures rather frequently (2). Some of the zygomatic arch fractures may be in the form of component of the mid-face fracture(2). The others in the form isolated fractures in the arch only results from localized forced landing on the face laterally and having relatively less impact3. Isolated zygomatic arch fractures comprise about 10% of all zygomatic fractures(2). When not treated properly, the arch fractures may lead not only to various cosmetic deformities related to skeletal structure of the face but also to functional disorders resulting from the pressure on the coronoid process(3). There are a lot of studies which related to demographic distribution of isolated arcus zygoma fractures in the literature. In these studies it is reported that fractures were more common in males than females (3). Also in our study almost whole of fractures were occur in males. Although in same investigation, falls were mostly observed etiological factor, in our study determined that violence is the most occur etiological factor (3). The left side was more frequently involved than the right side, but the reason for this is unknown (3). Classification of the fractures greatly facilitates the surgeon’s choice of the treatment and also classification facilitates that it constitutes a common terminology among surgeons and enables communication, too (3). Facial fractures have recently been classified in fine detail according to computed tomographic findings (3). However, recently there has been no classification of the generally encountered isolated zygomatic arch fractures to provide guidance for treatment (2, 3). To make detailed classification of zygomatic fractures in various shapes, which does not exist in the literature, and to form an algorithm for treatment was attracted attention (3). Yamomato et al (3). therefore classified the fractures into 5 types according to the degree of displacement and loss of bone contact (3). This classification is useful to determine the treatment method and need for fixation. Yamomato et al (3) reported that in type I fractures with no displacement, reduction is not necessary. In type III and IV fractures with displacement without bone contact at 1 or 2 fractured sites, good reduction can be obtained, Volume 2 ∙ Number ∙ 3 ∙ 2009

ARCUS ZYGOMA FRACTURES Rezzan TANRIKULU et al.

although there may be less stability after reduction, however, in type V fractures, open reduction may be needed to reduce comminuted bone fragments together with fixation or stabilization to maintain their alignment (3). Honig and Merten (5). were classified the fractures which called by their names, on the below: (HM) class I is defined as an isolated tripod fracture, HM class II as an isolated stick fracture of the arch, and HM class III is a combined fracture of the malar bone and the zygomatic arch. Researchers determined that open reduction is mandatory in class III fractures (HM) (5). The reduction status was evaluated by axial x- ray film and classified into 4 types: excellent, good, fair, and poor . Excellent reduction was achieved mostly in type II fractures (5). Fracture of the zygomatic arch is usually treated using blind methods. As the fracture lines cannot be visualized directly in closed reduction, digital exploration and crepitus noise or conventional radiographic imaging are used clinically as a guide to reposition the fragments. Successful closed reductions are often difficult (6). Postoperative radiographs are often the only way to assess the adequacy of the reduction. Intraoperative assessment of the zygomatic arch is very important in achieving adequate repositioning (6). The correct alignment of the zygomatic arch indicates the proper position of the zygomatic bone and ensures adequate prominence of the lateral midfacial aspect6. Gulicher et al studied on estimate the value of ultrasonography as an intraoperative repositioning control in the treatment of the zygoma fractures and they concluded that ultrasonography rapid and easy perform, and is recommended, and is intraoperative visualizing tool (6). And also in the literature it was reported that the use of ultrasonography, portable fluoroscopy and C-arm may have the advantage of intraoperative evaluation of the reduction status (6, 7). The treatment for isolated zygomatic arch fractures depends on the degree of displacement (1). Usually, fractures with significant displacement need reduction (1). There are several methods to reduce zygomatic arch fractures such as intraoral approach (Keen modality), hook traction (percutanous approach), temporal approach by Gillies’s and open reduction (4). Some investigator reported that Gillies’s approach has been most frequently used modality (1, 3). These researches are be opinion that the Gillies’s method offers several advantages, such as easy execution under local anesthesia, little possibility of facial nerve damage or direct trauma to the globe, and no visible Page 84


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scars within the hairline (3). Yamomato et al reported that in your department, reduction by the Gillies’s method was the first choice, because the procedure can be performed consistently and the results are satisfactory (3). In the same way we also treated delayed fracture successfully. Other methods of reduction via lateral eyebrow incision and by a transcutaneously inserted hook have also been reported (5). Recently, intraoral approaches have become the preferred choice due to the advantage of leaving no visible scars. Of these 9 patients, 5 patients were treated by Keen method and complete anatomic reduction was obtained. Besides these methods hook traction method is frequently used (1, 4) and we also preferred hook traction for 3 of 9 patient. Yamomato et al (3) reported that with laceration of the buccal skin below the affected zygomatic arch, hook reduction was also chosen (3). There were no laceration for our patient. In 1 case 2-3 mm incision was made on arcus zygoma. For the others percutanous approach was performed. In 2 of 3 cases that treated with hook traction method overcorrection was occurred. Some researches reported that intermaxillary fixation (IMF) is not necessary after zygomatic arch reduction. However in communicated fractures other researches reported that open reduction and fixation may be necessary (4). We preferred, IMF to prevent possible relapse for 5-7 days. We are in opinion of that IMF for short period doesn’t make discomfort and useful to prevent relapse. Recently endoscopic reduction and fixation method has been used for zygomatic fractures (7). The use of an endoscope may be helpful to perform reduction and fixation inside the optical cavity via a small incision. And also endoscopeassisted zygomatic arch realignment and fixation allow anatomic repair without sustaining the drawbacks of extensive access incisions (7, 8).

ARCUS ZYGOMA FRACTURES Rezzan TANRIKULU et al.

Additionally desired results are not guaranteed for this technique (10). Besides this modality tchrakeal tube and foley catheter is used for treatment of arcus zygoma fracture in literature (11, 12). References 1. Fonseca, R; J, Oral and Maxillofacial Surgery, ,Chapter 6, Volume 3, 1st edition, p. 198-199. W.B Saunders Company 2000. 2. Ozyazgan I, Günay GK, Eskitaşçıoğlu T, Özköse M, Çoruh A; A new proposal of classication zygomatic arch fractures. J Oral and Maxillofac Surg. 2007;65: 462-469. 3. Yamamoto K., Murakami K., Sugiura T., Fujimoto M., Inoue M., Kawakami M. Ohgi K., Kirita T; Isolated Zygomatic Arch Fractures. J Oral Maksillofac Surg 2007. 4. Kruger E; Oral and Maxillofacial Traumatology, Schilli W, Chapter 10, Volume 2, p. 172- 173. Quintessence Publishing Co., Inc., Chicago, Illinois 1986. 5. Honig JF, Merten HA. Classification system and treatment of zygomatic arch in the clinical setting. J Craniomaxillofac. Surg. 2004;15: 986-9. 6. Gulicher D, Krimmel M, Reinert S; The role of intraoperative ultrasonography in zygomatic complex fracture repair. Int. J Oral and Maxillofac. Surg. 2006;35: 224-230. 7. Badjate SJ, Cariappa KM. C-Arm for accurate reduction of zygomatic arch-a case report. Br Dent J 2005;10: 275-7. 8. M. Czerwinski and C. Lee, Traumatic arch injury: Indication and an endoscopic method of repair, Facial Plast Surg. 2004;20:231-238. 9. Carter TG, Bagheri S, Dierks EJ. Towel clip reduction of depressed zygomatic arch fracture. J Oral Maxillofac Surg. 2005;63:1244-54. 10. Manzon S, Choudhary N, Philbert R, Towel clip reduction of the depressed zygomatic arch fracture. J Oral Maxillofac Surg. 2006;64:1323-1326. 11. El-Haddy AM: The use of Foley catheter in isolated zygomatic arch fractures. Plast. Reconstr. Surg. 2005;116: 853-856. 12. Holmes S, Bridle C. Use of tracheal tube in isolated fractures of the zygomatic arch. Plast Reconstr. Surg. 2006;117:315.

Conclusions Another treatment modality towel clip reduction which created by Carter et al (9). Although the towel clip reduction of the depressed zygomatic arch fracture may be another simple technique in the armamentarium of surgical management of facial fractures, the unpredictability, limitations, and operator technique sensitivity can have an impact on the success of the operation where the procedure is applied (10). Manzon et al(10) reported that this technique is limited because it may only be used to reduce minimally displaced fractures. Volume 2 ∙ Number ∙ 3 ∙ 2009

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DENTO-ALVEOLAR INJURIES Hilal ALAN, Gulten UNLU, and Vedat TARI

A RETROSPECTIVE STUDY OF DENTO-ALVEOLAR INJURIES IN DIYARBAKIR, TURKEY Hilal ALAN1, Gulten UNLU2*, Vedat TARI3, Serkan AGACAYAK3 1. MsC, PhD, DDS, Researcher, Dicle University, Faculty of Dentistry, Department of Oral and Maxillofacial Surgery, Diyarbakir / TURKEY. 2. Prof.Dr., Dicle University, Faculty of Dentistry, Department of Oral and Maxillofacial Surgery, Diyarbakir / TURKEY. 3. MsC, DDS, Researcher Assist., Dicle University, Faculty of Dentistry, Department of Oral and Maxillofacial Surgery, Diyarbakir / TURKEY.

Abstract The purpose of this retrospective study was to determine the occurrence and the type of traumatic dental injuries managed by the University of Dicle, Faculty of Dentistry, Department of Oral and Maxillofacial Surgery. A total of 539 patients were seen for dental injuries with 1597 tooth injuries. During a period of 22 years, among 539 patients with dentoalveolar injuries, 300 (55.7 %) had been involved in falls, 144 (26.7%) resulted from traffic accidents, 47 (8.7%) from assaults, 29 (5.3%) from animal kicking, 11 (2%) from accidents at work, 5 (1%) in accidents that were sports related, 4 (0.6%) from accidents of an unspecified nature. This study shows that dental injury rates and patterns in a Diyarbakır-Turkey region are similar to other populations. (Journal of International Dental and Medical Research 2009; 2: (3), pp. 86-88) Keywords: Dentoalveolar trauma, Dental injuries, Retrospective Study. Received date: 21 August 2009 Introduction Traumatic injuries to the teeth are among the most serious oral health problems in active children and adolescents. Dental injuries are particularly important because of the critical sensory, communicative, gustatory and psychosocial functions of the teeth and the mouth. Unlike injuries to other parts of the body, fractures of the crowns of the teeth do not heal or repair. Dental trauma may have an uncertain prognosis; damage may appear at the time of the injury, and traumatic luxation can eventually result in pulpal necrosis (1). Dental injuries are caused by motor vehicle accidents, falls, sports accidents, interpersonal violence, animal kicking and iatrogenic reasons (during dentoalveolar surgery or using laryngoscopy for general anesthesia) and occurs in isolation or in association with facial injuries or multi-system injuries (2-8). The first successful attempt to classify dental injuries was made by Ellis and Daury in *Corresponding author: Prof. Dr. Gulten UNLU, DDS Phd Dicle University, Faculty Of Dentistry, Department of Maxilofacial Surgery, 21280 Diyarbakir, TURKEY. Tel: +90 412 248 81 01-3486 Fax: +90 412 248 81 00 E-mail: gunlu@hotmail.com

Volume 2 ∙ Number ∙ 3 ∙ 2009

Accept date: 14 October 2009 1970. This was expanded by the World Health Organization in 1978 and more recently by Andreasen and Andreasen in 1994. The classification of dental injuries may be divided into four categories: Injuries to the hard dental tissues and the pulp; injuries to the periodontal tissues; injuries to the supporting bone and injuries to the gingiva and oral mucosa (1). The pattern of injury depends upon the site, direction and energy of impact and the resilience of the periodontal structures surrounding the tooth. Injuries to the teeth such as concussion, subluxation, lateral luxation, and intrusion tend to occur if the lips cushion the impact or the force is distributed over several teeth. If the force hits the teeth directly, fracture of a crown, displacement of a tooth, and penetrating lip wounds are more likely. When the force is indirectly transferred to the teeth by an axial blow to the chin, fractures of crowns plus or minus root fractures are more likely, and the possibility of fracture of the mandible, luxation of the temporomandibular joint (TMJ), and cerebral injury must not be overlooked (6). The purpose of this study was to determine the frequency, outcomes and risk factors for dental injury in all patients’ age groups presenting at the Department of Oral and Maxillofacial Surgery at the Faculty of Dentistry, University of Dicle in Diyarbakır, Turkey.

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Materials and Methods Information concerning age and sex distribution, etiology, types, place and extent of trauma as well as seasonal variations, time difference between traumatic injury and seeking of dental care and number of traumatic injuries was recorded retrospectively from 539 patients which treated in the Department of Oral and Maxillofacial Surgery, University of Dicle, Diyarbakır, Turkey between January 1986-January 2008 during 22 year period. Patient characteristics were analyzed using descriptive statistics. Comparisons were performed with χ2 tests.

DENTO-ALVEOLAR INJURIES Hilal ALAN, Gulten UNLU, and Vedat TARI

The overall monthly distribution of dental injuries is uneven, peaking in summer and declining in winter (Fig.2).

Results During a period of 22 years, among 539 patients with dentoalveolar injuries, 300 (55.7 %) had been involved in falls, 5 (1%) in accidents that were sports related, 144 (26.7%) resulted from traffic accidents, 47 (8.7%) from assaults, 29 (5.3%) from animal kicking, 11 (2%) from accidents at work, 4 (0.6%) from accidents of an unspecified nature (Fig.1).

Figure 2. Seasonal distribution of dento-alveolar injuries. The type of injuries was summarized in Fig 3. The most common dental injuries were exarticulation (avulsion) and followed by luxation.

Figure 3. Type of injuries. Figure 1. Causes of injury. Seventy percent of all injuries occurred in men and boys and thirty percent in women and girls, yielding a male-to-female ratio of 2.3:1. The age range was 1 years to 64 years (mean 13 years) and most common people in the 515 years old age group. Anterior maxillary teeth and especially maxillary central incisors were most common traumatized teeth. There were altogether 1597 injured teeth from the 539 trauma cases. In other word, one patient averagely had 3 teeth traumatized. It was also found that 70% of dental traumas were permanent teeth and 30% primary teeth. For both types of dentition, injuries to the upper anterior teeth were predominant. Volume 2 ∙ Number ∙ 3 ∙ 2009

Discussion We have evaluated the incidence and etiology of dentoalveolar trauma in patients over a 22 years period, from January 1986, to January 2008. Data were collected for medical history, symptoms reported or pathologic signs displayed by the patients, and results of clinical and radiologic examination. The records of patients were analyzed according to frequency of dental trauma in facial injuries, age and gender distribution, monthly and yearly distribution of injury, cause of injury, frequency and type of injury. Our results confirm that boys are more prone to injuries than girls. A possible explanation for this could be that girls mature earlier than boys, who also tend to be more involved in outdoor activities Page 87


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(2-9). The etiology of injuries in the present study was in agreement with earlier reports. The most common cause of injury was a fall. As a child learns to walk and run, the incidence falls increases because the development of co-ordination and mobility is immature (2,3,10). When young children fall they are less able to protect the face, and are likely to sustain soft tissue injuries to the lips, tongue and face (3,9). Older age groups are more likely to be involved in traffic accidents. This confirms previous studies (9). Studies performed Gassner shown that, play accidents were the prime cause of dental trauma and followed by sport accidents (4, 5). The cumulative monthly distribution of injuries peaked in the summer months and fell in the winter months. This has been observed in several other papers (2-4, 9). Traffic voyages are further more in the summer because of holiday and traffic accidents increase in the summer. All of these may be the reasons that dental injuries cases increase in the summer. Sae-Lim et.al (1995) were reported that 79% of dental trauma were permanent teeth and 21% primary teeth (8). This result is appropriate the result of ours. It is reported that in both primary and permanent dentitions, dental trauma was significantly more often inflicted on maxillary anterior teeth (8, 9). In the present sample was in agreement with earlier reports. Mostly investigators were reported that the most common type of trauma were luxation injuries (4, 5, 8, 9). In comparison with the findings of other studies, the most common type of injury is extrusion in this study. The most common type of treatment used in our clinic was arch bar splint (29%) and followed by wire composite splint (25%), wire splint (25%), and extraction (21%) (Fig.4).

DENTO-ALVEOLAR INJURIES Hilal ALAN, Gulten UNLU, and Vedat TARI

The teaching of injury epidemiology and injury prevention to health care workers should be improved. Population-based epidemiologic data on injuries must be used to reassess conventional wisdom about injuries and to target future efforts at prevention of dentoalveolar injuries. Conclusions In this article, we provide evidence that databases such as the one instituted at our hospital raise and establish awareness of causes of and reasons for trauma. This study shows that dental and facial skeleton injury rates and patterns in Diyarbakir-Turkey region are similar to other populations. In addition emergency medicine can play an important role in the initial management of tooth injuries. Therefore, further study with a dentist is recommended. Acknowledgements: The authors wish to thank Prof.Dr. Yusuf Çelik for analyzed statistics of the study. References 1. Birgen N, Inanıcı MA, Aliustaoglu S, The forensic evaluation of dental injuries in Istanbul, Turkey. Forensic Science International 1999;106: 37-7. 2. O’Neil DW, Clark MV, Lowe JW, Harrington MS, Oral trauma in children: A hospital survey. Oral Surg Oral Med Oral Pathol 1989;68:691-6. 3. Kotecha S, Scanell J, Monaghan A, Williams RW, A four year retrospective study of 1,062 patients presenting with maxillofacial emergencies at a specialist paediatric hospital. Br J Oral Maxillofac Surg 2008; 293-296. 4. Gassner R, Tuli T, Hachl O, Moreira R, Ulmer H, Craniomacillofacial trauma in children: a review of 3,385 cases with 6,06 injuries in 10 years. J Oral Maxillofac Surg 2004; 62:399-9. 5. Gassner R, Bösch R, Tuli T, Emshoff R, Prevelence of dental trauma in 6000 patients eith facial injuries. Oral Surg Oral Med Oral Pathol 1999; 87:27-6. 6. Dewhurst SN, Mason C, Robert J, Emergency treatment of orodental: a review. Br J Oral Maxillofac Surg. 1998;36:165-11. 7. Newland MC, Ellis SJ, Peters RK et al. Dental injury associated with anesthesia: a report of 161,687 anesthetics given over 14 years. Journal of clinical Anesthesia 2007;19:339-7. 8. Sae-Lim V, Tan HH, Yuen KW, traumatic dental injuries at the accident and emergency department of Singapore general hospital. Endod Dent Traumatol 1995;11:32-5. 9. Schatz JP, Joho JP, A retrospective study of dento-alveolar injuries. Endod Dent Traumatol 1994;10:11-4. 10. Love RM, Ponnambalam Y. Dental and maxillofacial skeletal injuries seen at the University of Otago School of Dentistry, New Zealand 2000-2004. Dental Traumatology 2008; 24: 170–7.

Figure 4. Type of Treatment. Volume 2 ∙ Number ∙ 3 ∙ 2009

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MUTHUPETTAI MANGROVE ENVIRONMENT P. Manivasagan et al

BACTERIAL COMMUNITY AND PHYSICO-CHEMICAL CHARACTERISTICS OF MUTHUPETTAI MANGROVE ENVIRONMENT, SOUTHEAST COAST OF INDIA Manivasagan P1*, Sivakumar K2, Thangaradjou T3, Vijayalakshmi S4, Balasubramanian T5 1. Research Scholar, CAS in Marine Biology, Annamalai University, INDIA 2. Senior Lecturer, CAS in Marine Biology, Annamalai University, INDIA 3. Lecturer, CAS in Marine Biology, Annamalai University, INDIA 4. Research Associate, CAS in Marine Biology, Annamalai University, INDIA 5. Professer & Director, CAS in Marine Biology, Annamalai University INDIA

Abstract A study on bacterial community and physico-chemical parameters of water and sediment samples in mangrove environment of Muthupettai, South east coast of India was carried out in 2006-07. Six stations in an around the mangrove sites were selected for sampling and the following parameters were recorded at monthly intervals atmospheric temperature, surface water temperature, sediment temperature, salinity, water pH, sediment pH, dissolved oxygen, nitrite, nitrate , total nitrogen, inorganic phosphate, total phosphorus, ammonia, silicate and the microbial quality like total heterotrophic bacteria (THB), total coliforms (TC), faecal coliforms (FC), pathogenic bacteria as Vibrio cholerae, V. parahaemolyticus, Escherichia coli, Salmonella spp., Shigella spp., Klebsiella spp., Streptococcus spp. and Pseudomonas spp. THB strains (six hundred and seventy) and Vibrio strains (one hundred and twenty five) were isolated from mangrove environment. When compared to nutrient distribution, water nutrients were consistently higher in the mangrove environment. Partially treated aquaculture waste water are having additional sources of nutrients. Although, the mangrove habitat has been demonstrated to possess self-cleaning properties, data obtained warms possible anthropogenic pollution in the mangrove area in near future if the present conditions prevailed for a long period. (Journal of International Dental and Medical Research 2009; 2: (3), pp. 89-99) Key words: Mangrove, characteristics.

Bacterial community,

Received date: 23 September 2009 Introduction Mangrove wetlands along the coastal zone act as barrier against cyclones, protect coastal erosion and provide good nursery ground for number of commercially important aquatic organisms (Janaki-Raman et al. 2007).In mangroves and coastal waters, factors related to water quality such as temperature, salinity, pH, dissolved oxygen and nutrients are particularly important for determining the biological factors and ecosystem functions (Paramasivam and Kannan, 2005). Many mangrove environments are receiving pollutants due to varied activities taking place

*Corresponding author: P. Manivasagan* Research Scholar CAS in Marine Biology Annamalai University / INDIA E-mail: manimaribtech@gmail.com

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Muthupettai, Nutrients, Physico-chemical

Accept date: 13 November 2009 around the mangrove areas and with the inflowing freshwaters, thus resulting in the degradation of water quality. Therefore, it is necessary to monitor their habitat characteristics and water quality (Paramasivam and Kannan, 2005). The abundance and distribution of total heterotrophic bacteria have a direct bearing on other forms of nutrients in different compartments of the environment. Microbial indicators have been used world wide to indicate if human wastes have contaminated a water body and they are the most common member of faecal coliforms which indigenous to the intestinal tract of human and other warm-blooded animals. The microbes more commonly utilized are those found in elevated concentrations in human feces. The typical indicators used in the mangrove environment include total coliforms, faecal coliforms, Esherichia coli and Enterococci. In this paper, we report on the bacterial community and physico-chemical characteristics of Muthupettai mangrove environment, Southeast coast of India. Page 89


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Materials and methods Study area Muthupettai mangroves (Lat. 10o 25’N; Long. o 79 39’E) situated 400km south of Chennai lies along the south east coast of India. It has total area of 6800ha in which the water spread area covers approximately 2720 ha (Fig.1).

St 1

St 2

St 3

St 4

St 5

St 6

Figure 1. Map of Muthupettai mangrove environment showing different locations. It has two specialized habitats viz. mangroves and lagoon. Many tributaries of the river Cauvery delta such as Paminiyar, Koraiyar, Kilaithangiyar, Kandankurichanar and Marakkakoraiyar flow through Muthupettai and nearby villages and form a lagoon before they enter into the sea, Bay of Bengal. Avicennia marina is the dominant mangrove species in Muthupettai and accounts for nearly 95% of the vegetative cover. The sampling areas of Station 1 (Aquaculture discharge area); Station 2 (Sethuguda); Station 3 (Lagoon); Station 4 (Sellimunai); Station 5 (Sea mouth region) and Station 6 (Open Sea) were selected for the present study. Surface water and sediment samples at six stations were collected at monthly intervals during April (2006) to March (2007) and transported to the laboratory by keeping them in ice box and processed within 24 hours and microbial analysis were carried within 4 hours. Physico-chemical analyses Initial measurements on temperature (digital thermometer), pH (pH Scan 1 Tester-Eutech Instruments) and salinity (Refractometer Atago F/mill 8901) of the water samples were made onboard and dissolved oxygen was estimated by the modified Winkler’s method described by Volume 2 ∙ Number ∙ 3 ∙ 2009

MUTHUPETTAI MANGROVE ENVIRONMENT P. Manivasagan et al

Strickland and Parsons (1972). Concentration of water nutrients such as nitrite (NO2), nitrate (NO3), total nitrogen, inorganic phosphate, total phosphorus (PO4), ammonia (NH4) and silicate (SiO3) were analyzed by following the methods of Strickland and Parsons (1972) and APHA (1995). Sediment pH was measured according to Chattopadhyay (1980).Parsons correlation coefficient was carried out for understanding the interrelationships between various physico-chemical parameters using SPSS-10. Microbiological analysis Total heterotrophic bacteria The total heterotrophic bacteria (THB) count was determined on Zobell marine agar medium using the spread-plate technique, in triplicate. After 48h incubation the colonies were counted. Morphologically diverse colonies were isolated from each plate and sub-cultivated several times on Zobell marine agar medium. Isolated strains were stored in a fridge at 4oC in marine agar slants (prepared with filtered seawater adjusted to 20% salinity with distilled water) (Sousa et al. 2006). Vibrio counting and isolation Vibrio spp. were counted and estimates of numbers made using the compendium of microbiological methods (Downes and Ito, 2001). The colonies from each Thio sulphate citrate bile salt sucrose (TCBS) plate were selected and subcultured in marine agar medium for isolation of the strains (Sousa et al. 2006). Enumeration of enteric pathogens Total enteric pathogens were enumerated by adopting the membrane filter technique using XLD Agar (Xylose Lysine Deoxycholate Agar) medium recommended for the selective isolation of enteric pathogens especially Salmonella sp., Shigella sp. and Klebsiella sp. species. After 48 hours of incubation, colonies appeared with a typical pink, pink to red colours of Shigella sp. and with yellow colours of Salmonella sp. and with black colours of Klebsiella sp. were isolated. Enumeration of total coli forms Total coliforms (TC) were counted by standard membrane filter (MF) methods using MacConkey Agar. The MF method involves filtering a sample through 0.45μg pore size membrane filter (47mm diameter membrane, Fisher, Pittsburg, PA) that retains the bacteria (Tomoyuki et al. 2004) were impregnated in the Petri plates. Appropriate volumes of samples were filtered and placed on MacConkey Agar and incubated at 41oC for 24h. Page 90


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Enumeration of faecal coliforms The same MF method was used for enumerating faecal coliform utilized a modified MFC agar (Tomoyuki et al. 2004) and incubated at 44+0.5oC for 24h and colonies with various shades of blue were counted as faecal coliforms. Enumeration of Escherichia coli M7 Hr FC Agar was used as a selective medium for E. coli isolation by employing membrane filtration procedure. Enumeration of Enterococci sp. Enterococci sp were also analyzed by membrane filter (MF) methods using MEnterococcus Agar. After 24 hours of incubation at 41oC, the colonies of S. faecalis appeared with maroon colour were counted. Enumeration of Pseudomonas aeruginosa Cetrimide Agar was used as selective medium for the isolation of P. aeruginosa by employing membrane filtration procedure. After 48 hours of incubation at 37oC, the colonies of P. aeruginosa appeared with luxuriant yellow colour were enumerated. Biochemical analysis All the strains were identified using biochemical analysis according to Bergey’s manual of determinative bacteriology (Buchanan and Gibbons, 1974) without modifications. The bacteria isolated from the TCBS plates (selective for Vibrios), were further identified by biochemical tests described by Tison (1999). Results Physico-chemical characteristics Atmospheric temperature, surface water temperature, sediment temperature, salinity, water pH, sediment pH, dissolved oxygen, nitrite, nitrate, total nitrogen, inorganic phosphate, total phosphorus, ammonia and silicate values are shown in Fig. 2 – 15. In general there is only very little spatial variation in most of the physicalchemical parameters recorded during the study outing to their closer geographical location. However, there is a cleat temporal variations in most of these parameters. Specifically speaking increasing amount of water nutrients recorded during the monsoon season correlating with land run off and higher river water inflow. Regarding correlation study between the Volume 2 ∙ Number ∙ 3 ∙ 2009

MUTHUPETTAI MANGROVE ENVIRONMENT P. Manivasagan et al

parameters of water, station 1 water temperature showed a significant negative correlation with dissolved oxygen (r = -857). Nitrate showed a significant positive correlation with total phosphorus at p =0.01 level. In station 2 atmospheric temperature showed a significant positive correlation with salinity (r = 915). Ammonia showed a significant negative correlation with inorganic phosphate at p =0.01 level. In station 3 pH showed a significant negative correlation with nitrite (r = 842). While silicate exhibited a positive correlation with total nitrogen and significant at p =0.01 level. In station 4 total nitrogen showed a significant positive correlation with inorganic phosphate (r = 959). Silicate showed a significant positive correlation with total phosphorus at p =0.01 level. In station 5 atmospheric temperature showed a significant negative correlation with total nitrogen (r = -802) and ammonia showed a negative correlation with total phosphorus at p =0.01 level. In station 6 salinity showed a significant negative correlation with dissolved oxygen (r = -840). Nitrate showed a positive correlation with inorganic phosphate at p =0.01 level. Microbiological analysis Total heterotrophic bacteria, Vibrio cholerae, Vibrio parahaemolyticus, Escherichia coli, Salmonella spp., Shigella sp., Klebsiella sp., Streptococcus sp., Pseudomonas sp., total coliforms and faecal coliforms values are shown in Fig. 16-37. Regarding correlation study between the parameters of water and sediment, station 1 total heterotrophic bacteria showed a significant positive correlation with V. parahaemolyticus (r = 946). Escherichia coli showed a positive correlation with Klebsiella sp. and significant at p =0.01 level. In station 2 V. cholerae showed a significant positive correlation with total coliforms (r = 958). Salmonella spp. showed a positive correlation with Pseudomonas sp. and significant at p =0.01 level. In station 3 Shigella sp. showed a significant positive correlation with faecal coliforms (r = 922). The Klebsiella sp. exhibited a positive correlation with Pseudomonas sp. and significant at p =0.01 level. In station 4 faecal coliforms showed a significant positive correlation with Pseudomonas sp. (r = 963). Shigella sp. showed a positive correlation with faecal coliforms and significant at p =0.01 level. In station 5 V. cholerae showed a significant positive correlation with Salmonella spp. (r = 959). Klebsiella sp. showed a negative correlation with faecal coliforms and significant at p = 0.01 level. In station 6 V. parahaemolyticus showed a significant positive correlation with total coliforms (r = 921). E. Page 91


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coli showed a positive correlation with Shigella sp. and significant at p = 0.01 level. Identification of total heterotrophic bacteria Totally 670 strains were isolated based on colony morphology from THB plates for identification of genus (Table 1). The majority of the isolated strains from station 2 and station 5 were identified. The lowest number of isolates was identified from station 6. Identification of Vibrio spp. One hundred and twenty five strains isolated on the Vibrio TCBS selective medium were characterized phenotypically (Table 2). The number and names of species isolated from the TCBS plates. The greatest number of different species isolated was found at station 1. The second greatest number of species isolated was detected at station 2 and 5. The lowest number of isolates was identified from station 6.

Table 1. Heterotrophic bacteria identified from mangrove environment.

Table 2. List of Vibrio spp. Identified from mangrove environment. Discussion Study attempted to correlate the native physico-chemical parameters with bioavailability of different antibiotic resistant bacteria and its population in the mangrove environment of Muthupettai, India. Temperature is one of the most Volume 2 ∙ Number ∙ 3 ∙ 2009

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important factor responsible for the regulation of physiological activities of microorganisms. In nature, each species has its own optimal temperature requirements for their growth as well as development (Paramasivam and Kannan, 2005). In the present study, the seasonal mean temperature was maximum in summer and minimum in monsoon. Temperature fluctuation was observed between 22.8 and 34.8oC. Slightly lower temperature was recorded at Station 2 and higher in station 4. All the stations recorded lower (0o/oo) salinity during the monsoon season than the other seasons reaching the maximum (43.1o/oo) during the summer when there is no fresh water flow in the rivers. Higher pH recorded (8.4) during the summer in the present study due to the removal of CO2 by the photosynthetic organisms and the lower pH observed (7.2) during the monsoon season. The first two factors encourage a heating of the water during the day, provoking evaporation and an increase in the salinity values and rain reduces the temperature, pH and salinity during monsoon period. The present study observed that the nitrite, nitrate and total nitrogen concentrations in water and sediment nitrogen in higher ranges (1.362µmol/l, 5.436µmol/l, 25.361µ mol/l and 8.59µg/g) respectively. Inorganic phosphate and total phosphorus concentration of water were observed in range between 0.36 to 1.23 µ mol/l, 1.71 to 5.36µ mol/l respectively and sediment phosphorus was in the range of 0.75 to 2.67µg/g. Regarding concentration of ammonia in water higher range in (0.077µ mol/l) monsoon season. The maximum concentration of reactive silicate (156.8µmol/l) was observed during the monsoon period and the minimum recorded during summer. The results were highly correlate with the results of Paramasivam and Kannan (2005). The increased level of nutrients in water and sediment during monsoon period due to land run off and freshwater inflow in the rivers through leaching from manured and fertilized agricultural soils, aquaculture discharge and sewage effluents from the surrounding environment. Higher THB population density of water (71×106 CFU ml-1) and sediments (75×107 CFU g-1 dry wt) was recorded from Station 2. Kathiresan (2000) has reported that dead organic matter in the sediments of the mangrove area favors the higher bacterial growth in this environment. The lower density (31×106 CFU ml-1) in water were recorded at station 6 and (39×107 CFU g-1 dry wt) in sediment were recorded at Station 5. This could be possible because this mangrove area is open in nature as the mangrove leaves takes more than 10 days to Page 92


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get decompose (Kathiresan, 2000) and more than 50% of the mangrove leaves were transported to other places before they get decomposed (Ajithkumar et al., 2006). In the present study, Higher V. cholerae and V. parahaemolyticus population density in water and sediment (41×103 CFUml-1) and (45×104 CFU g-1dry wt) were recorded from Station 1 with shrimp forms effluent pollution and indicators of sewage pollution. The lower density was recorded in the water (16×103 CFUml-1) and sediment (19×104 CFU g-1 dry wt) samples of Station 5. Higher V. parahaemolyticus population density in water and sediment (38×103 CFUml-1) and (42×104 CFU g-1dry wt) were observed from station 4 and station 6. The lower density was recorded in the water (12×103 CFUml-1) and sediment (16×104 CFU g-1 dry wt) samples of Station 5. The results were similar by Sousa et al, 2005. Higher total coliforms population density in water and sediment (63×105 CFUml-1) and (68×106 CFU g-1dry wt) were recorded from Station 6. The total coliforms are the most widely used indicators for the microbial pollution evaluation of fresh and marine waters, which suggest that coliforms don not survive in high salinities particularly under the sunlight. The lower density was recorded in the water (25×103 CFUml-1) and sediment (27×104 CFU g-1 dry wt) samples of Station 3. The effect of sunlight is a function of the water salinity which indicates that coliforms in seawater are subjected to a lower rate of survival. Total coliforms are poorly resistant microorganisms in the natural environment. Higher faecal coliforms population density in water and sediment (55×104 CFUml-1) and (58×105 CFU g-1dry wt) were recorded from Station 6. The presence of faecal coliforms in the seawater by the shore indicates that the source of pollution is likely to be domestic waste from the local sewage outfall, fresh water inflow and other unidentified sources of faecal pollution such as mangrove environment. The lower density was recorded in the water (21×103 CFUml-1) and sediment (23×104 CFU g-1 dry wt) samples of Station 3. Coliforms are poorly resistant microorganisms in the natural environment. However, the constant flow of domestic wastes produces high levels of pollution which permanently decreases the quality of seawater for recreational purposes. Similar results were observed by Hashim et al. 2005. Higher population density E. coli in water (32×103 CFUml-1) and sediment (35×104 CFU g-1dry wt) were recorded from station 5 and 1. A monsoon season resulted in the higher loading of E.coli and subsequent high E.coli in mangrove environment. In addition, higher loading of nutrients with runoff Volume 2 ∙ Number ∙ 3 ∙ 2009

MUTHUPETTAI MANGROVE ENVIRONMENT P. Manivasagan et al

could help the growth of E.coli, if any growth occurs. E.coli was present in sea water during all seasons in mangrove environment indicating that the water was contaminated by faecal material of humans or other warm-blooded animals and also indicates the potential for the presence of pathogenic organisms (Youn et al, 2002). The lower density was recorded in water (15×103 CFU ml-1) and sediment (17×104 CFU g-1dry wt) from station1 and 5. E.coli was lower in the summer season, which is lower loading of faecal material, fresh water inflow and the ecological condition of the mangrove environment. Similar results were observed by Youn et al. 2002. Higher population density in water (28, 29, 26×102 CFU ml-1) and sediment (30, 31, 28×103 CFU g-1dry wt) samples were recorded from station 2, 3 and 5. The monsoon season observed in the higher loading of faecal materials and subsequent high freshwater in mangrove environment. Seawater quality criteria proposed by the world health organization and other international foundations are usually suggested enteric forms as microbiological pollution indicator. The combination of several indicators is likely to present a global picture of water quality. The lower density was recorded in water (11, 10, 6×102 CFU ml-1) and sediment (13, 13, 9×103 CFU g-1dry wt) from station 3 and 4. In summer season observed in the low of level of nutrients, changes in environmental parameters and decrease the faecal contamination. The results were similar by Arone and Walling, (2007). In present study, higher population density in water (49×102 CFU ml-1) and sediment (51×103 CFU g-1dry wt) samples were recorded from station 2. Among feacal pollution indicators is a higher survival of streptococci in the marine environment. Sunlight inactivation of culturable cells of enterococci generally required 2-3 times the isolation for culturable cells of faecal coliforms. A decrease of inactivation rates of enterococci at lower temperatures was also observed. The lower density was recorded in water (24×102 CFU ml-1) and sediment (26×103 CFU g-1dry wt) from station 1. The presence indicated a recent contamination and showed that the mangrove was effectively subjected to a continuous animal pollution which decreased or disappeared in summer and similar results were reported by (Bouchriti et al. 1992). Higher population density in water (35×102 CFU ml-1, and sediment (39×103 CFU g-1dry wt) samples were recorded from station 5 with regarded as organisms of a faecal origin. Especially higher amount of Pseudomonas sp. were found once in the bottom sediment of the higher number of organic Page 93


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substance, phytoplankton, PO4, NO3, adsorbed to clay is a factor advantageous for proliferation and survival of these bacteria in the mangrove environments (Niewolak and Opieka, 2000). The lower density was recorded in water (16×102 CFU ml-1) and sediment (19×103 CFU g-1dry wt) from station 3 with less polluted in summer season. The results were similar by (Niewolak and Opieka, 2000). Totally 680 strains of total heterotrophic bacteria were identified in genus levels at different stations, which are affected by monsoon season heavy fresh water inflow, agricultural discharges, shrimp effluent pollution with indicators of sewage pollution and this result suggests that perhaps other anthropogenic sources of pollution are present and influencing the microbial communities at all sites. The majority of the isolated strains from station 2 and 5 were identified, which are affected by aquaculture pond discharge water in mangrove environment. Similar results were observed by Hashim et al. (2005). The lowest number of isolates was identified at station 6. In the station, fresh water inflow, pollution sources and aquaculture effluent were low in marine environment. One hundred and twenty five strains isolated were identified in species level at different stations. The greatest number of different species isolated was found at station 1. The second greatest number of species isolated was detected at station 2 and 5. In addition to the isolates which are important pathogens for humans and aquatic animals, the genus Vibrio also includes species involved in nutrient cycling such as V. harveyii not to mention others capable of breaking down chitin, aromatic polycyclical hydrocarbons which are extremely toxic for the environment (Thompson et al, 2004). Similar results were recorded by Sousa et al. 2006.

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Ajithkumar, T.T., Thangaradjou, T., L. Kannan 2006. Physico-chemical and biological properties of the muthupettai mangrove in Tamilnadu. J. Mar. Biol. Ass. India, 48: 131-138. American Public Health association (ALPH), 1995. Standard methods for the Examination of Water and Waste Water, 14th ed. APHA, Inc, Washington, DC. Arone, R.D, Walling, J.P., 2007. Waterborne pathogens in urban watersheds. Journal of Water and Health.149-162. Balasubramanian, R., Kannan, L., 2005. Physico-chemical Characteristics of the Coral Reef Environs of the Gulf of Mannar Biosphere Reserve, India. Int. J. Ecol. Environ. Sci. 31(3): 273-278. Bouchriti, N, El Marrakchi, A., Fahim, A., 1992. The microbiological contamination of an oyster growing area in Morocco: The Oualidia lagoon. Hydroecol. Appl, 4(2): 189-202. Buchanan, R.H, Gibbons, N.E., 1974. Bergey’s manual of determinative bacteriology. 8th ed. The Williams & Wilkins company, Baltimore. Pp.529-550.

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Chattopadhyay, S. 1980. Hydrology and landuse in the littoral tract of deltaic West Bengal. Geographical Review of India, 42 : 177 180. Downes, M.P, Ito, K., 2001. Compendium of Methods for the Micobiological Examination of Foods, fourth ed. APHA, Washington, DC, 676 p. Hashim, A, Al-Sayed, H. Eassam, M, Ghanem, M, Kareema, Saleh. 2005. Bacterial community and some physico-chemical characteristics in a subtropical mangrove environment in Bahrain. Marine pollution Bulletin, 50: 147-155. Janaki-Raman, D, Jonathan, M.P., Srinivasalu, S., Armstrong-Altrin, J.S, Mohan, S.P., Ram-Mohan, V., 2007. Trace metal enrichments in core sediments in Muthupet mangroves, SE coast of India: Application of acid leachable technique. Environmental Pollution 145:245-257. Kathiresan K, (2000) A review of studies on Pichavaram mangrove, southeast India. Hydrobiologia, 430: 185 – 205. Niewolak, S., Opieka, A., 2000. Potentially Pathogenic Microoganisms in water and Bottom Sediments in the Czarna Hancza River. Polish Journal of Environmental Studies, 9(3):183-194. Paramasivam, S, Kannan, L., 2005. Physico-chemical Characteristics of Muthupettai Mangrove Environment Southeast Coast of India. International journal of Ecology and Environmental Sciences 31(3):273-278. Sousa, O.V, Macrae, A., Menezes, F.G.R., Gomes, N.C.M., Vieira, R.H.S.F., Mendonca-Hagler, L.C.S., 2006. The impact of shrimp farming effluent on bacterial communities in mangrove waters, Ceara, Brazil. Marine Pollution Bulletin, 52: 17251734. Strickland, J. D. H, Parsons, T.R., 1972. A practical hand book of seawater analysis. Bull. Fish. Res. Bd. Can., 167:310pp. Thompson, F.L, Iida, T., Swings, J., 2004. Biodiversity of Vibios. Microbiol. Mol. Biol. Rev. 68, 403-431. Tison, D.L, 1999. Vibrio. In: Murray, P.R., Baron, E.J., Pfaller, M.A., Tenover, F.C., Yolken, R.H. (Eds), Manual of Clinical Microbiology, Seventh ed., pp. 497-506(1773 p). Tomoyuki Shibata, Helena M. Solo-Gabriele, Lora E. Fleming, Samir Elmir, 2004. Monitoring marine recreational water quality using multiple microbial indicators in an urban tropical environment. Water Research, 38: 3119-3131. Youn-Joo An, Donald H.Kampbell, Peter Breidenbach G, 2002. Escherichia coli and total coliforms in water and sediments at lake marinas. Environmental Pollution 120:771-778.

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STRUCTURE OF FILIFORM AND FUNGIFORM PAPILLAE Ayfer AKTAS, and Yusuf NERGIZ

THE HISTOLOGICAL STRUCTURE OF FILIFORM AND FUNGIFORM PAPILLAE OF THE PUPPY RATS WHICH BORN FROM THE RATS GIVEN EGF AFTER SIALOADENECTOMY Ayfer AKTAS1*, Yusuf NERGIZ2 1. Assist. Prof. Dr. Department of Histology and Embryology, Medical School, Dicle University, 21280, Diyarbakir / TURKEY 2. Prof. Dr. Department of Histology and Embryology, Medical School, Dicle University, 21280, Diyarbakir / TURKEY

Abstract After the complete removal of submandibular gland surgically, morphological alterations, and the effects of EGF given orally on these alterations were investigated. In the study, 30 adult female SpraqueDawley rats were used, which were divided equally into three groups: Control group; sialoadenectomy group ; sialoadenectomy + epidermal growth factor group . Following a three week period of recovery, the rats were mated. On days 16-19 th of pregnancy, a total of 5 microgram EGF was given to the animals in sialoadenectomy + epidermal growth factor group with orogastric tube as 1.25µg daily to each animal. The fetuses born following pregnancy were kept to grow up until the 28th day . The pupy rat of 28 days in all groups were taken, and then they were sacrificed. Tongues of all puppy rats were fixed in a 10 % neutral buffered formalin solution. The paraffin sections obtained through routine histological methods were stained with Hematoxylen–Eosine , Methylen blue- Basic Fuchsin, Hemotoxylen–Van Giesson, and were examined under light microscope. A significant increase in the keratinization; an elongation and bifurcation at the ends of the filiform papillae of the sialoadenectomy group rats were observed . The fungiform papilla and the taste buds were noticed to be larger than that of the control group, and a decrease was detected in the cells population that constitutes the taste buds. Bifurcation and hyperkeratosis in the filiform papillae of the sialoadenectomy + epidermal growth factor group rats.The morphological appearance of fungiform papillae and taste buds was observed to be similar to that of fungiform papillae of the control group. (Journal of International Dental and Medical Research 2009; 2: (3), pp. 100-104) Keywords: Rat, EGF, Sialoadenectomy, Lingual Papillae. Received date: 05 October 2009 Introduction The tongue papillae were first identified by Malpighi in 1664 (1). The Taste buds found in humans and mammals are the receptors for the perception of taste and in rats there are three types of taste bud-bearing tongue papillae, which are fungiform papilla, circumvallate papilla, and foliate papilla (2). While the fungiform papillae are on the anterior portion of the tongue and the foliate papillae on the rear-lateral edges, the circumvallate papilla, which is singular, is placed

*Corresponding author: Dr.Ayfer AKTAS Department of Histology and Embryology Medical School, Dicle University 21280, Diyarbakir / TURKEY Phone:+90 4122488001-4123 Fax: +90 4122488435 E-mail: aaktas@dicle.edu.tr, fuldic@gmail.com

Volume 2 ∙ Number ∙ 3 ∙ 2009

Accept date: 22 Nowember 2009 more posteriorly on the midline of the tongue (3). The circumvallate papilla which is located in the midline of the dorsum lingua is circumscribed from the rear and lateral sides with a horseshoe-shaped, incompletely enclosed trench, that is left and right trenches do not merge in the anterior part (4). Farbman and Mbeine (5), observed the formation of a mushroom shaped fungiform papilla with mechanical and gustatory functions in the tongues of 15-day fetuses. It was announced that every single fungiform papilla has a single taste bud on it in a ratio of 99% in rats as in hamsters. Miller and Preslar (6) divided the tongue into two; and they declared that more than 50% of the papilla was located in the edges of the tongue. While the average papilla density was 3.4/mm2 in the first 4 mm of the tongue, it decreased to 1.3/mm2 in the 4-17 mm range and no papilla was present in the dorsal line. There were more papillae in the right half of the tongue with respect to the left one. Page 100


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The filiform papillae of rats have a mechanical effect and their histological structure was demonstrated by Farbman (4). However, there are limited amount of work dealing with the different aspects of filiform papillae. The filiform papillae start to develop in the last days of embryonic life. The length and the base width of the papillae show an increase with age. A soft keratin layer on the front and lateral faces and a hard keratin layer on the rear faces of filiform papillae can be distinguished. Filiform papillae have a pointed end tended backwards and the shape of a cone getting thinner going up (6, 7). Material and method 1. Animals In this study 30 adult female SpragueDawley rats weighing about 250-300 g and provided from the Practice and Research Center of Medical Sciences in Dicle University were used. The female rats were divided into three groups in equal numbers (n=10) as control(C), sialoadenectomy (SX), and sialodenectomy + Epidermal Growth Factor (EGF), respectively. 2. Surgical Procedures Control Group (C): Female rats of this group were allowed to mate without any operation. The first day of the pregnancy was determined by the microscopic examination of vaginal smear. On the 28th day following their birth, tongues of all puppy rats were fixed in a 10% neutral bufferd formalin solution. Sialodenectomy (SX): Female SpragueDawley rats were put under general anesthesia by intramuscular administration of Ketamine + Xylazine. A transversal incision was performed at the neck region and the submandibular glands were totally extracted (8). For three weeks following the operation it was waited for the EGF level to drop. Then the test animals were mated, and the first day of the pregnancy was determined through the microscopic examination of vaginal smears. On the 28th day following their birth, tongues of puppy rats were put into a 10% neutral bufferd formalin solution for the histological examination. Sialodenectomy + Epidermal Growth Factor (SX+EFG): As in Group 2, the submandibular glands of female rats were removed by a sialodenectomy operation. The Volume 2 ∙ Number ∙ 3 ∙ 2009

STRUCTURE OF FILIFORM AND FUNGIFORM PAPILLAE Ayfer AKTAS, and Yusuf NERGIZ

rats of this group were let to mate after three weeks following the operation and the starting day of their pregnancy were determined again by microscopic examination of vaginal smears. Along the four days between 16th and 19th days of pregnancy, a 1.25 μg of EGF (Human Recombinant EGF, sigma) per subject was administered daily to pregnant rats by orogastric probe, making a sum of 5 μg per subject in total (8). On the 28th day following their birth, tongues of puppy rats were fixed in a 10% neutral buffered formalin solution. 3. Histological Method On the 28th day following the birth, tongues of puppy rats of all three groups were dissected and cut into two from the line determining the anterior 2/3 portion and the posterior 1/3 portion. Then the samples were separately put into 10% neutral buffered formalin solutions. Sections with thicknesses of 4-6 μm were taken transversally and coronally from the paraffin blocks, which were obtained by routine paraffin treatment, with the aid of a sliding microtome. After passed through an ethanol series and xylol, the sections were stained with Haematoxylin-Eosin (HE), Haematoxylin-Van Giesson, and Methylene Blue + Basic Fuchsin, and then examined under a light microscope, and microphotographs were taken. Results The findings of tongue samples are summarized as follows: 1. Control Group No pathology could be showed on the panoramic views of fungiform and filiform papillae (Fig. 1).

Figure 1. Control Group: The panoramic view of the filiform (thin arrow) and fungiform (thick arrow) Page 101


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STRUCTURE OF FILIFORM AND FUNGIFORM PAPILLAE Ayfer AKTAS, and Yusuf NERGIZ

papillae on the dorsum lingua. (Methylene blueBasic fuchsin, original magnification X 40) 2. Sialoadenectomy Group (SX) A significant increase in the keratinization; longitudinal elongation of apikal spines in several fungiform papillae in SX animals had apical spines of filiform papillae of the SX group rats were observed (Fig. 2a).

Figure 3a. SX+EGF group: The panoramic view of the filiform papillae (thin arrow)(Methylene blue-Basic fuchsin, Original magnification X 40). The morphological appearance of fungiform papillae and taste buds was observed to be similar to that of fungiform papillae of the control group (Fig. 3b).

Figure 2a. SX group: Note the apical spines at the tips of filiform papillae (thin arrow) (H-E. original magnification X 40). The fungiform papilla and the taste buds were noticed to be larger than that of the control group, and a decrease was detected in the cells population that constitutes the taste buds (Fig. 2b).

Figure 3b. SX+EGF group: Fungiform papilla (thick arrow) and taste bud (t) with normal appearances( Methylene blue-Basic fuchsin, Original magnification X 40). Discussion

Fig. 2b: SX group: The hypertrophied fungiform papillae (thick arrow) and taste buds (t) in the apical epithelium. (Methylene Blue-Basic fuchsin, original magnification X 80) 3. Sialoadenectomy + Epidermal Growth Factor Group (SX+EGF) Bifurcation and hyperkeratosis in the filiform papillae of the SX+EGF group rats (Fig. 3a). Volume 2 ∙ Number ∙ 3 ∙ 2009

Shinzaburo and co-workers reported that the EGF concentration in the saliva of the mice and rats who had undergone sialoadenectomy was considerably diminished and because of this the recovery period of tongue wounds was elongated. It is known that the orally administered EGF afterwards, which does not increase the EGF level of portal and peripheral blood, enhances the healing process of tongue scars. Besides it was reported to cause mouth dryness in mice, as the amount of saliva in the mouth was decreased by the extraction of main Page 102


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saliva source, i.e., submandibular gland. In mice, the inadequacy of EGF is the major cause of retardation observed in the recovery of wounds. It was discovered that the recovery rate of wounds reached usual levels when exogenous EGF was introduced. As quite wellknown, the tongues of mice are covered with a statified squamous epithelium; and since the EGF found in saliva cannot be absorbed from the lingual epithelial under normal circumstances, it is reported not to affect the basal epithelial cells. However, if the lingual epithelial is impaired due to certain chemical or mechanical agents, the EGF in the saliva can easily access the basal epithelial cells and contributes to the healing of wound. There was, however, a noticeable increase in keratinization of the lingual epithelian and increased infiltration of inflammatory cells within the mucosa SX groups. The healing process of wounds in mice with sialoadenectomy was reported to exhibit a two-day lag with respect to normal healthy mice (9). In our study, while the fungiform papillae of the control group exhibit a natural appearance, an elongation and bifurcation at the tips was witnessed in those of the SX group, together with a significant increase in the keratinization (Fig 2a-b). Epitelium and thinging in fungiformis papilla atrophy of taste buds. İncrease in the surface epithelium spines and covering a large potion of the papilla. However bifurcation and increase in the keratinization could not be detected in the fungiform papillae of the SX+EGF rats thanks to orally administered EGF (Fig 3a).

The morphology of the fungiform papillae undergoes a change and the number of taste buds is declined in the rats tretment to sialodenectomy, which makes itself clear in the supplementation of EGF as feedback. The healthy development of taste buds in the fungiform papilla is the proof of the important role that EGF plays. The exogenous administration of EGF to the rats subjected to sialoadenectomy causes an increase in the volumes of fungiform papillae. SX group when compared with the effects depending on the faste buds EGF in the epithelial layer and the thickness of the prominent observed. Although some researchers suggest not only a parallel between the distribution of fungiform Volume 2 ∙ Number ∙ 3 ∙ 2009

STRUCTURE OF FILIFORM AND FUNGIFORM PAPILLAE Ayfer AKTAS, and Yusuf NERGIZ

papillae located in the anterior part of the tongue and their taste buds but also the presence of a taste bud in the apical of each papilla, there are some other researchers claiming that not all the fungiform papillae have a taste bud on them. The results obtained from our study concerning to fungiform papillae contradicts with the works of Beidler (13). The tongue papillae of the rats are arranged both from the anterior to posterior and also to the middle in series in the tounge. Mistretta and et al pointed that the arrangement of the fungiform papillae were in series of longitudinally and diagonally (medial-lateral) in thetounges of rat fetuses in their study (10). Farban has indicated that the distrubition of the fungiform papillae of the rat fetuses were similar with the adult rat papillae (11). Therewithal, Vich and et al also reported that the arrangement of the papiilae on the surface of the tounge were parallel with the branches of the lingual nerve (12). Cano and co-workers investigated the role of sialoadenectomy in the maturation and survival of taste buds and they discovered that it affected the development of specific taste bud cell types. These observations lead the researches to think that some other factors also in the saliva must have been responsible for the sustenance of the natural taste function and morphology, but these factors could not be detected yet (14). After the experimental work performed on the taste buds of the rat fungiform papillae, MorrisWiman et al. found that in the rats subjected to sialoadenectomy, the taste buds were atrophied and they were composed of only a few cells. Besides, their surface areas were rather diminished with respect to the sham group. When the EGF was administered orally afterwards, an improvement on the taste buds of the fungiform papillae was noticed; in fact they acquired an appearance identical to that of the sham group. In our study, the average diameters of the taste buds of control group and SX group fungiform papillae were measured as 122 μm and 88 μm, respectively. There was a decrease in the number of cells that make up the taste buds. The sole reason for the fungiform papillae and their taste buds being hypertrophied is thought to be the extensive elimination of EGF source by the total extirpation of submandibular glands by sialodenectomy. İn the tongue of sialoadenectomized rats, fungiform papillae often had spines reminiscent of the apical spines of filiform papillae EGF supplementation restored epithelium and papillae. Our findings concerning the taste buds of the fungiform papillae are consistent with the results of the studies carried out by MorrisWiman and co-workers (15). Page 103


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Conclusions The structural changes observed in the fungiform and filiform papillae due to sialoadenectomy display a significant recovery as a result of the orally administered EGF. References 1. Bradley R.M, Stern IB The develepment of the human taste bud during the foetal period. J Acat 1980;130(1):25-32. 2. Fish HS, Malone DD, Richter CP The anatomy of the tongue of the domestic Norvay rat. I. The skin of the tongue, the various papillae, their number and distribution. Anat Rec 1994; 89: 429440. 3. Iwasaki S, Yoshizawa H, Kawahara I. Study by Electron Microscopy of the morphogenesis of three types of lingual papilla in the rat. Anat Rec 1997 ;247: 528-541. 4. Farbman AI The dual pattern of keratinization in filiform papillae of rat tongue. J Anat 1970; 106(2) :232-242. 5. Farbman A., Mbiene JP Early development and innervation of taste bud–bearing papillae on the rat tongue. The J. Comparative Neurol 1991;304:172-186. 6. Miller IJ, Preslar AJ Spatial distribution of rat fungiform papillae. Anat Rec 1974;181:679-684. 7. Reynolds G Handbook of Histological Techniques, 2nd edition, Department of Histopatholog, London 1990; 5-36. 8. Whitehead MC, Kachele DL Distrubution of taste and general sensory nerve endings in fungiform papilla of the hamster J Comp Neurol 1994; 340:515-530. 9. Noguchi S, Ohba Y, Oka T Effect of salivary epidermal growth factor on wound healing of tongue in mice. Am J Physiol 1991; 260: E620- E625. 10. Misteratta C. M Topographical and histological study of the devoloping rat tongue, palate and taste buds. Third Symposium on Oral Sensation and Perception: The mouth of the infant, J. F. Bosman, ed. Thomas, Spring field 1972; 163187. 11. Farbman AI Electron microscope study of developing taste bud in rat fungiform papilla. Dev Biol 1965; 27(2) :263-272. 12. Vij SR Development of the nerve supply to the human tongue. Acta Anat 1972; 81: 466-477. 13. Beidler LM, Smallman RL Renewal of cells within taste buds. The J Cell Biol 1965; 27: 263-272. 14. Cano J. Roza C, Rodriguez EL Effects of selective removal of the salivary glands on taste bud cells in the vallate papilla of the rat. Experientia 1978; 34 :1290-1291. 15. Morris- Wiman J, Sego R, Brinkley L, Dolce C The Effects of sialoadenectomy and exogenous EGF on taste bud morphology and maintenance. Chem. Senses 2000; 25: 9-19.

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HYPOLIPIDEMIC EFFECT Jeyanthi K.A, and Mary Violet Christy .A

HYPOLIPIDEMIC EFFECT OF CITRULLUS COLOCYNTHIS SEED POWDER IN ALLOXAN INDUCED DIABETIC RATS Jeyanthi K.A.1*, Mary Violet Christy A.2 1. Department of BioChemistry, Thanthai Hans Roever College, Perambalur Tamilnadu / INDIA. 2. Department of Zoology, Govt Arts College, Ariyalur, Tamilnadu / INDIA.

Abstract This study was undertaken to investigate the effect of Citrullus Colocynthis on blood glucose and some other Lipid biochemical parameters in alloxon induced diabetic rats. The aqueous extract treatment(300mg/kg body weight) was given for 22 days. Effect of this plant on the level of Glucose, total cholesterol, tri-glycerides, phospholipids, free fatty acids, LDL and HDL cholesterol and the enzyme activities like lipase, acetylcoA carboxylase, HMG coA reductase in serum and liver were evaluated. The Plant and Glibenclamide treated rats significantly reduce the elevated levels of Sugar, Cholesterol and other lipids in serum and liver. So these results indicate that Citrullus Colocynthis posse’s hypolipidemic effect. (Journal of International Dental and Medical Research 2009; 2: (3), pp. 105-109) Keywords: Hypolipidemic, citrullus colocynthis, alloxan, glibenclamide. Received date: 20 October 2009 Introduction Diabetes Mellitus is the heterogeneous metabolic disorder characterized by altered carbohydrate, lipid and protein metabolism (1). India has today become the diabetic capital of the world with over 20 million diabetics and this number is set to increase to 57 million by 2025 (2). Along with hyperglycemia and abnormalities in serum lipids, diabetes is associated with micro and macro vascular complications, which are the major causes of morbidity and death. Management of diabetes without any side effects is still a challenge to the medical system. This leads to increasing demand for natural products with antidiabetic activity and less side effects. Therefore investigation from the medicinal plants has become more important because India has a rich history of using various potent herbs and herbal component for treating diabetes. Many Indian plants have been investigated for their beneficial use in different types

*Corresponding author: Mrs. K.A.JEYANTHI Professor & Head Department of BioChemistry Thanthai Hans Roever College Perambalur – 621 212 Tamilnadu / INDIA Mobile : 9994223240 E-mail: jeyanthi.ka@gmail.com

Volume 2 ∙ Number ∙ 3 ∙ 2009

Accept date: 16 November 2009 of diabetes and reported in numerous scientific journals.

Materials and Methods Plant Material: The fresh plant seeds of citrullus colocynthis was collected from perambalur area, Tamilnadu, was identified and authenticated from Botanist Dr. V.Kumaresan, Department of Biotechnology, Thanthai Hans Roever College, Perambalur. The seeds were dried and powdered in grinding mill. Preparation of Extracts: The Seeds were pulverized in grinder. 200gm of powdered seeds was poured in 400ml of distilled water for 1 hour at room temperature with continuous shaking. The mixture was filtered using sterile gauze. A fresh filtrate was used for treatment. Animals: Male Albino rats weighting about 150-200 gm obtained from the Eaisma Institue, Karur were used for the study. The rats were kept in the animal house at room temperature of 15 – 30 0 , fed with commercial food ad libitum and had free access to water. Animal studies in the work have been strictly performed as per the institutional of Animal Ethical Committee, Govt of India, and New Delhi.

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Induction: Alloxan Monohydrate was used to induce diabetes mellitus in normoglycemic rats. Animals were allowed to fast for 16 hours and were injected intraperitonially with freshly prepared alloxan monohydrate in normal saline in a dose of 120mg/kg body weight (3). Blood Glucose was measured after 72 hours by one touch glucometer and it was confirmed by testing for Glucosuria using Glucose indicator sticks. Rats showing fasting blood glucose levels > 200 mg/dl were selected for the study. Experimental Design: The animals were divided into 5 groups of six animals in each group. Group 1: Normal Control. Group 2: Normal Control + Plant treatment (300mg/kg body wt). Group 3: Diabetic Control. Group 4: Diabetic Control + Plant treatment. Group 5: Diabetic Control + Glibenclamide treatment ( 0.5mg/kg body wt). Biochemical Parameters:

Group

Glucose (mg/dl)

Glycosylated Haemoglobin (mg/g/dl) 4.15 ± 0.15 4.23 ± 0.20a 10.23 ± 0.38b* 4.95 ± 0.15cd* 5.90 ± 0.88c*

HYPOLIPIDEMIC EFFECT Jeyanthi K.A, and Mary Violet Christy .A

The Blood Glucose level was determined by the method of sasaki et al.,(4). The Hemoglobin level was analysed by Cyanmethemoglobin method of drabkin and Austin. The Glycosylated Haemoglobin was measured by the method of Nayak and Patabiraman. Plasma insulin level was assayed by ELISA kit using human insulin as standard. The free fatty acid was determined by the method of Hron and Menahan (5-7). The cholesterol was estimated by Zak’s method. Triglycerides was determined by the method of Rice (8). The Phospholipids was estimated by Bartlette Method (9). HDL and LDL were measured by the method of Burstein and Scholnick (10). The Lipase, HMGcoA reductase and acetylcoA carboxylase was determined by enzymatic kit method. Statistical Analysis: The values are expressed as mean ± Standard Deviation for six rats in each Group. All other data were analysed with SPSS/16.0 student software. Hypothesis testing method included oneway analysis of variants( ANOVA) followed by Post Hoc testing performed with least significant difference (LSD) test. The P value of less than 0.05 was considered to indicate statistical significance.

Haemoglobin (g/dl)

Insulin

Urine Sugar

Normal Nil 89.33 ± 2.16 11.3 ± 0.90 92.16 ± 6.49 a a a Normal + CC.Aqt 93.50 ± 5.57 Nil 10.0 ± 0.48 87.00 ± 19.43 Diabetic Control 228.00 ± 5.66b* 7.73 ± 0.57b* 48.18 ± 12.64b* +++ Diabetic + CC.Aqt 92.60 ± 2.16cd* + 9.8 ± 0.29cd* 83.56 ± 2.39cd* Diabetic + + 109.17 ± 16.25c* 12.66 ± 1.65c* 91.66 ± 6.25c* Glibenclamide Table 1. Effect of Citrullus Colocynthis extract on the levels of Glucose, Glycosylated Haemoglobin, Haemoglobin, Insulin and urine sugar in Serum of alloxan Diabetic rats.

Group

Cholesterol LDL HDL (mg%) (mg%) (mg%) Normal 156.16 ± 2.63 79.33 ± 1.21 61.66 ± 1.47 Normal + CC.Aqt 156.50 ± 3.39a 72.83 ± 1.32a* 62.16 ± 3.52a b* b* Diabetic Control 330.50 ± 31.86 178.16 ± 1.60 18.66 ± 2.16b* Diabetic + CC.Aqt 169.66 ± 6.34cd* 84.00 ± 6.13cd* 58.36 ± 5.98cd* c* c* Diabetic + Glibenclamide 161.66 ± 21.60 79.01 ± 1.41 56.38 ± 1.23c* Table 2. Effect of Citrullus Colocynthis extract on the levels of serum, cholesterol ,LDL, HDL level in control and experimental rats.

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HYPOLIPIDEMIC EFFECT Jeyanthi K.A, and Mary Violet Christy .A

Group

Triglycerides free fatty acids Phospolipids (mg%) (mg%) (mg%) Normal 119.50 ± 1.04 10.75 ± 3.06 167.17 ± 29.32 Normal + CC.Aqt 118.01 ± 2.16a 10.40 ± 3.23a 167.39 ± 27.38a Diabetic Control 197.83 ± 11.51b* 23.33 ± 0.65b* 335.10 ± 30.82b* cd* cd* Diabetic + CC.Aqt 117.50 ± 1.87 10.41 ± 2.27 203.33 ± 15.88cd* Diabetic + Glibenclamide 105.83 ± 17.44c* 11.46 ± 1.33c* 190.36 ± 21.67c* Table 3. Effect of Citrullus Colocynthis extract on the levels of serum, triglycerides, free fatty acids, phospolipids level in control and experimental rats. Group

AcetylcoA HMGcoA reductase carboxylae (IU/L) (IU/L) Normal 0.76 ± 0.35 7.25 ± 0.75 1.01 ± 0.25 a a Normal + CC.Aqt 0.77 ± 0.44 7.36 ± 0.78 1.08 ± 0.40a Diabetic Control 2.90 ± 0.04b* 3.46 ± 1.04b* 0.32 ± 0.11b* cd* cd* Diabetic + CC.Aqt 1.08 ± 0.30 7.20 ± 0.62 1.00 ± 0.52cd* Diabetic + Glibenclamide 0.96 ± 0.23c* 7.81 ± 0.78c* 1.13 ± 0.36c* Table 4. Effect of Citrullus Colocynthis extract on the levels of Serum ,lipase, acetylcoA carboxylase and HMGcoA reductase in control and experimental group of rats. Group Normal Normal + CC.Aqt Diabetic Control Diabetic + CC.Aqt Diabetic + Glibenclamide

Lipase (IU/L)

phospholipids (mg/dl)

triglycerides (mg/g/dl)

166.83 ± 29.21 a* 169.80 ± 27.63 b* 90.10 ± 61.53 cd* 161.00 ± 41.97 c* 165.56 ± 21.84

123.80 ± 11.32 a* 120.61 ± 9.68 b* 210.10 ± 5.57 cd* 131.75 ± 6.31 c* 127.01 ± 4.56

free fatty acids (mg/dl) 9.75 ± 3.06 a* 8.56 ± 2.02 b* 4.10 ± 1.30 cd* 8.61 ± 2.29 c* 9.88 ± 1.67

HDL (mg/dl)

LDL (mg/dl)

40.66 ± 9.60 a 40.83 ± 4.91 b* 23.16 ± 4.99 cd* 41.16 ± 7.52 c* 42.46 ± 3.32

66.83 ± 12.75 a* 69.16 ± 24.16 b* 146.50 ± 6.31 cd* 76.66 ± 4.13 c* 71.41 ± 6.44

Table 5. Effect of Citrullus Colocynthis extract in liver phospholipids, triglycerides, free fatty acids, HDL and LDL cholesterol levels in control and experimental rats. Group

lipase acetylcoA HMGcoA reductase (IU/L) (IU/L) carboxylase (IU/L) Normal 0.70 ± 0.31 7.01 ± 0.68 0.90 ± 0.25 Normal + CC.Aqt 0.76 ± 0.44a* 7.28 ± 0.82a* 1.02 ± 0.50a* b* b* Diabetic Control 0.15 ± 0.02 2.58 ± 0.74 0.33 ± 0.15b* Diabetic + CC.Aqt 0.65 ± 0.45cd* 7.46 ± 0.91cd* 0.82 ± 0.40cd* c* c* Diabetic + Glibenclamide 0.72 ± 0.31 7.16 ± 0.79 0.95 ± 0.37c* Tab. 6 Effect of Citrullus Colocynthis extract in liver lipase, acetylcoA carboxylase, HMGcoA reductase activities in control and experimental rats. Values are given as mean ± SD of six rats from each group Values are statistically significant at *P<0.05 a-> Normal + Plant treated rats were compared with Normal Rats. b-> Diabetic Rats were compared with Normal Rats. c-> Plant treated Diabetic Rats were compared with Diabetic Rats. d->Glibenclamide treated Diabetic Rats were compared with Diabetic Rats.

Results Treatment with citrullus colocynthis aqueous extract on blood glucose level, insulin, Hb, HbA1C and Glycogen level are depicted in Table-1. The rats exposed to alloxan developed diabetes as evident from the significant elevation in blood glucose and HbA1C as compared to normal control Volume 2 ∙ Number ∙ 3 ∙ 2009

rats. The administration of plant extract produced the significant reduction in blood glucose and HbA1C level. The insulin and Hb levels in diabetic control rats were decreased significantly compared to normal rats. In Diabetic treated groups(4 and 5) insulin and Hb levels are increased. Table-2 shows the levels of total cholesterol, LDL and HDL levels of serum in control and diabetic Page 107


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rats. The results revealed that there was a significant increase in total cholesterol and LDL level in diabetic rats. But the HDL level was reduced in diabetic control. The administration of the plant extract and the drug treatment shows the levels were reaches the near normal values. The levels of triglycerides, free fatty acids and phospholipids are depicted in Table-3. The elevated levels of TG, FFA and phospholipids in diabetic rats were reduced after the treatment. Table-4 and Table-6 shows the changes in the activities of lipase, acetylcoA carboxylase and HMGcoA reductase in serum and liver of control and experimental rats. The activity of lipase was increased in both serum and liver. In rats treated with the plant extract there was a reduction in the lipase activity. The reduced activities of acetylycoA carboxylase and HMGcoA reductase in diabetic control reach the normal value after the treatment of the plant extract. Table-5 shows the effect of phospholipids, TG, FFA, HDL and LDL levels in liver of control and experimental rats. The elevated levels of triglycerides and LDL levels were reaches the normal after the plant treatment. The Phospholipids, FFA and HDL level were decreased significantly in diabetic control rats. Treatment with citrullus colocynthis, the level was significantly increased and reaches the near normal value. Discussion Diabetes Diabetes Mellitus is possibly the world largest growing metabolic disease, and as the knowledge on the heterogeneity of this disorder is advanced, the need for more appropriate therapy increases (11). Traditional plant medicines are used throughout the world for a range of diabetic complication. The study of such medicines might offer diabetic complication. The study of such medicine might offer a natural key to unlock a diabetogenist’s pharmacy for the future. In animals, in can be induced by partial pancreatectomy or by the administration diabetogenic drugs such as alloxan, streptozotocin and anti-insulin serum (12). Alloxan, a beta cytotoxin causes a massive destruction of beta cells of the islets of langerhans resulting in reduced synthesis and release of insulin (13). It is well established that sulphonyl ureas produce hypoglycemia by increasing the secretion of insulin from pancreas. The antidiabetic effect of citrullus colocynthis aqueous extract could be linked to more than one mechanism, it includes the stimulation of Volume 2 ∙ Number ∙ 3 ∙ 2009

HYPOLIPIDEMIC EFFECT Jeyanthi K.A, and Mary Violet Christy .A

beta cells and subsequent release of insulin and activation of the insulin receptors. The plants have an antihyperglycemic action, may be by potentiation of pancreatic secretion of insulin, which was clearly evidenced by increased level of insulin in diabetic rats, treated with plant extract. In this context, a number of other plants have also been reported (14). HbA1C was found to increase in patients with diabetes mellitus to about 16% (15) and the amount of increase is directly proportional to the fasting blood sugar level (16). The citrullus colocynthis reduces the elevated HbA1C level in diabetic rats. In this study, we have also observed an increase in total cholesterol, TG and FFA in alloxan induced diabetic rats. Hyperlipidemia is a recognized consequence of diabetes mellitus (17). Diabetes induce hyperlipidemia is attributable to excess mobilization of fat from the adipose tissue due to the under utilization glucose(18). The ability of aqueous extract of citrullus colocynthis reduces the lipid level in diabetic rats. Cardio vascular disease and stroke are the major complication in diabetes. This is due to increased LDL cholesterol, which further undergoes oxidative modification in the presence of free radicals leads to atherosclerosis and vascular damage. A decrease in LDL with raised HDL cholesterol plays an important role against these complications. Insulin plays a key role in the metabolism of lipids apart from its regulation of carbohydrate metabolism. During diabetes, enhanced activity of lipase increases lipolysis and releases more fatty acids into the circulation (19). In this study we have observed an increase in lipase in alloxan induced rats. After the treatment the level was reduced. AcetylcoA carboxylase is the key enzyme whose induction in beta cells may contribute to glucolipotoxicity. It catalyses an essential step in fatty acid synthesis in lipogenic tissues, this is usually a minor pathway of glucose usage in beta cells (20). However malonylcoA, the immediate product of this enzyme may play a signaling role in glucose stimulated insulin secretion (21). In alloxan induced diabetes, which causes abnormal metabolism of ketone bodies and is known to result in decreases of acetylcoA carboxylase (22). After the plant and drug treatment the levels were significantly reaches the normal. HMGcoA reductase is a rate limiting enzyme in cholesterol synthesis. STZ induced diabetes in rats results in a marked decrease in the HMGcoA reductase in the liver (23). The role of insulin in promoting the synthesis of this enzyme in the liver Page 108


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and the serum has been appreciated , although other work (24) has indicated that increases in enzyme during diabetes. In our study the reduced HMGcoA reductase was increased after the plant treatment.

21.

22.

Conclusions 23.

From our study it is concluded that the aqueous extract of citrullus colocynthis seed powder exhibit hypolipidemic activity. This effect may be due to the presence of tannin, saponin, and flavanoids and other constituents of that plant.

24.

HYPOLIPIDEMIC EFFECT Jeyanthi K.A, and Mary Violet Christy .A

acetylcoA carboxylase gene expression in a pancreatic beta cell line (INS-1). J.Biol.Chem. Vol-268 : 18905-18911 Dasilva Xavier G, Varadi A, Ainscow E, Rutter GA(2000) Regulation of gene expression by glucose in pabcreatic beta cells via insulin secretion and activation of phosphatidyl inositol-3-kinase. J.Biol.Chem. Vol-275 : 36269-36277 Pape ME, Lopez – Casillas F, Kim KH. (1988). Physiological regulation of acetylcoA carboxylase gene expression. Effect of diet, diabetes and lactation on acetylcoA carboxylase mRNA. Arch.Biochem.Biophys. Vol-267 : 104-109 Young NL, Saudek CD, Craw Ford SA (1982). J.Lipid Res. Vol23:266-275 Nakayama H and Nakagawa S (1977). Diabetes. Vol-26 : 439-444.

References 1. Mutalik. S, Sulochana B, Chetana M, Udupa N, Uma devi P (2003). Preliminary studies on acute and sub acute toxicity of an antidiabteic herbal preparation. Dianex. Indian. J. exp. Biol. Vol – 41: 316-320 2. Sridhar GR (2000). Diabetes in India. Snapshop of a panorama. Current. Sci. Vol-83: 791 3. El-Demerdesh.F.M, Yousef.M.I, Abou E1-Nag.N.I(2005). Biochemical study on the hypoglycemic effects of onion and garlic in alloxan induced diabetic rats. Food Chem.Toxicol.Vol-43:57-63. 4. Sasaki T, Matsy S and Sonal A (1972). Effect of acetic acid concentration on the color reaction in the O-toluidine boric acid method for blood glucose estimation. Rinsh Kagaku Vol-1 : 346-353. 5. Drabikin DL and Austin JM (1932) Spectrophotometric constants for common hb derivatives in human, dog and rabbit blood. J.Biol.chem.Vol-98:719-733. 6. Nayak SS and Pattabiraman TN(1981) A New calorimetric method for the estimation of glycosylated hb.Clin.Chim. Acta Vol-109: 267274. 7. Hron WT, Menahan LA (1981). A sensitive method for the determination of free fatty acids in plasma. J.Lip.Res. Vol-18:663665 8. Rice EW (1970). Triglycerides in Serum. In standard methods in clinical chemistry (eds) Roedrick P and Donal RP. Academic Press, New york. P : 215 9. Barlette (1959). Phosphorous Assay in column chromatography. J. Biol. Chem Vol-234 : 466-468. 10. Burstein M, Seholnick HR (1972). Precipitation of Chylomicron and VLDL from human serum with sodium lauryl sulphate. Life Sci. Vol11 : 177-184. 11. Baily CJ, Flatt PR (1986). Antidiabetic Drugs and New Developments. Ind.Biotech. Vol-6 : 139 – 142 12. Carvalho EN, Ferreira LM (2003). Experimental Model of Induction of Diabetes Mellitus in rats. Acta.Cirurgica Brasileira. Vol-18 : 60-64 13. Lazarrow A (1954). Alloxan Diabetes and the mechanism of betacell damage by chemical agents. Experimental Diabetes. Oxford : Blackwell scientific publications. PP – 49-70 14. Kaleem M, Asif M, Ahamed QU, Bano B (2006). Antidiabetic and Antioxidant activity of annona squamosa extract in STZ induced diabetic rats. Singapore. Med.J. Vol – 47 : 670-675 15. Koeing.R, Peterson.C.M, Jones.R.L, Sandek.C, Lehrman.M, Lerami.A(1976). Correlation of glucose regulation and HbA1C in DM.N.Engl.J.Med. Vol-295:417-420. 16. Jackson RL, Hess RL, England JD (1979). HbA1C values in children with overt diabetes maintained in varying degree f control. Diabetes Care. Vol –2 : 391-395. 17. Pushparaj PN, Tan BKH and Tan CH (2001). The mechanism of hypoglycemic action of the semipurified fraction of Averrhoa Bilimbi in STZ induced diabetic rats. Life.Sci.Vol-70 : 535-547 18. Krishnakumar K, Augusti KT and Vijayammal PL (2000). Hypolipidemic effect of Salacia oblanga wall root bark in STZ diabetic rats. Med.Sci. Vol-28 : 65-67 19. Agardh CD, Bjorgell and Nilson EP (1999). The effect of tolbutamide on lipoproteins and hormone sensitive lipase. Diabetes.res.clin.Pract. Vol- 46 : 99-108 20. Brun T, Roche E, Kim KH, Prentki M (1993). Glucose regulates

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THE EFFECT OF 900 MHZ RADIOFREQUENCY (RF) RADIATION ON SOME HORMONAL AND BIOCHEMICAL PARAMETERS IN RABBITS. Beyza KARADEDE1, Mehmet Zulkuf AKDAG2*, Zeki KANAY3, Ayse BOZBIYIK4 1. Yeditepe University. Faculty of Dentistry, Istanbul / TURKEY. 2*.Department of Biophysics, Medical Faculty of Dicle University, Diyarbakir / TURKEY. 3. Department of Physiology, Veterinary Faculty of Dicle University, Diyarbakir / TURKEY. 4. Erbakir Science High School, Denizli / TURKEY.

Abstract In this study, the effect of cell phone frequency 900 MHz radiofrequency (RF) radiation on rabbits’ some hormonal and biochemical parameters was investigated. 14 male, adult, New Zealand albino rabbits were separated into two groups as control and experimental (n=7). For the experimental group, rabbits exposed to 900 MHz frequency RF radiation 2 h per day (7 days in a week) for 90 days. Control group rabbits’ procedure was the same as experimental group except the exposure of RF. Before the exposure, at first and third month of exposure; Glucose, Creatinin, Na, K, Cl, Calcium, Phosphorus, Total protein, Albumin, Globulin and Magnesium levels and T3, T4, TSH, LH, FSH, Testesteron, Estradiol, Prolactin, Cortisol, ACTH and Growth Hormon levels were determined from rabbits’ blood which was taken from V.Marginalis in their right ear. There was not any significant difference in neither hormonal nor biochemical parameters between experimental and control groups before exposure (p>0.05). At first month of RF exposure; there was not any significant difference in biochemical parameters (p>0.05). However, T4, Testesterone, Estradiol, Cortisol and ACTH levels increased but this increasing was not stastistically significant (p>0.05). After third month exposure, it was found that there was not any significant difference in biochemical parameters (p>0.05), but there was an increase at T4, Cortisol and ACTH levels and only increasing in ACTH levels was stastistically significant (p<0.05). It was concluded that 900 MHz RF exposure did not change some important biochemical and hormonal parameters except ACTH levels after 90 days exposure. (Journal of International Dental and Medical Research 2009; 2: (3), pp. 110-115) Key words: Mobile phone, Radiofrequency Radiation. ACTH, FSH,LH, Glucose, Albumin Received date: 23 June 2009 Introduction The exponential increase in the use of mobile phones is raising major concerns on possible consequences of the interaction between radiofrequency (RF) and biological systems (1). Also some of the international institutes and groups have tried to determine the possible risks by establishing the limited estimations of this radiation (2,3). They have been using calculated or observed

*Corresponding author: Dr. Mehmet Zulkuf Akdag Department of Biophysics, Medical Faculty of Dicle University, 21280, Diyarbakir-TURKEY Tlf: +90-412-2488001 Fax : +90-412-2488440 E-mail: akdag@dicle.edu.tr, mzakdag@gmail.com

Volume 2 ∙ Number ∙ 3 ∙ 2009

Accept date: 18 October 2009 thermal effects to determine these limitations, but do not consider possible effects of chronic or repeated nonthermal exposures (4). However, nonspecific neurovegetative syndromes such as physical and neural asthenia, sleep disorders, humoral changes, headache and myalgia have been observed in males exposed to chronic low intensity RF radiation (5). It was shown with animal studies that exposure to RF radiation had effects on the endocrine or the nervous systems and especially the thyrotropin (TSH) secretion (5). In addition, Navakatikian and Tomashevyskaya have determined inhibition in testesteron and insuline secretion in rats which are exposed to low intensity microwaves (6). Seze et al. reported that one month of intermittent exposures to RF radiation from a cellular telephone does not induce a long-lasting or cumulative effect on the hormone secretion rate of the anterior pituitary gland in humans (5). Page 110


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Braune et al. found that RF radiation emitted from mobile phone did not affect to the serum norepinefrin, epinefrin, cortisol and endotelin amounts of healthy people (7). In their study Rajkovic et al. have exposed to low-frequency electromagnetic fields to male rats during three months and afterwards they investigated the hormonal and functional structures of thyroid tissues (8). The results obtained from this study showed that exposure of low-frequency electromagnetic fields to male rats for 3 months have caused morphofunctional alteration on thyroid tissues. Mann et al. have investigated the effects of pulsed high-frequency electromagnetic fields that emitted from circular polarized antenna, on neuroendocrine system of healthy peoples (9). The hormones investigated in this study are Growth Hormone (GH), Cortisol, Luteinizing Hormone (LH) and melatonin hormone (9). The results obtained in this study determine that low intensity highfrequency electromagnetic fields emitted from mobile phone causes no effects on hormone secretion other than only a slight increase in cortisol formation (9). It has been reported that radio frequency radiation at mobile phone frequency exposed to rodents and human has changed many parameters related to stress. Stagg et al. who investigated this potential relationship, studied the effect of 1.6 GHz puls-modulated RF radiation on Fischer 344 rats’ Adrenocorticotropin (ACTH), corticosterone and brain ornithine decarboxylase (ODC) levels (10). At the end of the study, these investigators had reported that puls-modulated RF fields (up to 5 W/kg SAR level) did not indicate any stress response related to ACTH, corticosteroid and brain ODC levels. Radon et al. reported that the salivary concentrations of melatonin and cortisol did not change after 900 MHz RF exposure in healthy male students (11). The contradictions of the studies done on both human and experimental animals relating to the effect of RF radiation emitted from mobile phones on endocrine system, directed us to make such a study. In the present study, the results obtained from the long term chronic exposures will enlighten the contradictions about the potential effects of low intensity RF radiation. The main aim of our study is to investigate whether the RF radiation emitted by mobile phones, which their usage rapidly increase both in our country and in the world, has any effects on endocrine system and some biochemical parameters, and to help to enlighten the contradictions about the subject.

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THE EFFECT OF RF RADIATION Beyza KARADEDE et al

Materials and Methods Subjects and Animal Care The experiments were performed on 14 adult male New Zealand rabbits obtained from Medical Science Application and Research Center of Dicle University, aged 3- 4 months, weighing 2500- 3500g, and fed with standard pelleted food (TAVAS Inc. Adana, Turkey) and tap water. The animals were kept in 14/10h light/dark environment at constant temperature of 22 ± 3°C, 45 ± 10% humidity and one rabbit in one cage. 14 rabbits were separated into two groups such as control (n=7) and experimental (n=7). All animal procedures were in agreement with the principles of laboratory animal care and the rules of Scientific and Ethics Committee of Dicle University Health Research Center (Protocol Number; 2005/29). 900 MHz RF Exposure and Experimental Method Rabbits are placed in the radial mode, in cages (31×17.5×17) cm in size with opening and closing wood in the top, the front part bended and has 7 cm diameter circular opening that rabbits neck portion can pass through (Figure 1).

Figure 1. Experimental setup. RF radiation obtained from 900 MHz frequency RF generator (GSM Simulator 900PM10 type, Everest Comp., Adapazarı, Turkey) exposed to 7 rabbits in experimental group for 90 days two hours a day, 7 days a week. The antenna of RF generator is placed in a plastic cylindrical line so that all rabbits can expose to RF equally (Figure 1). The properties of RF signal obtained from the RF generator are: Frequency: 900 MHz, Output Power: 2 W, modulation frequency 217 Hz, pulse width: 577 µs. The power density of RF radiation obtained from the antenna were detected with an electromagnetic field meter (EMR-300, Narda, Pfullingen, Germany) on different regions of rabbit (Figure 1). For control group, the rabbits were placed into the wooden Page 111


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THE EFFECT OF RF RADIATION Beyza KARADEDE et al

cages (2 h/day/7day in a week for 90 days), same procedure were applied to the rabbits, except that RF generator was turned off. To determine the effects of RF radiation on some biochemical and hormonal parameters of rabbits, the blood samples of experimental and control groups collected three times from marginal ear vein with intravenous canulla depended of rabit’s size. The blood samples were taken before, immediately after first and third months of RF radiation exposure. Serum was removed from blood via centrifugation for 5 minutes at 6500 rpm. Determination of Biochemical Parameters For the biochemical parameters used in the study; Glucose, Creatinine, Na, K, Cl, Calcium, Phosphorus, Total Protein, Albumin, Globulin and magnesium values were determined by using the enzymatic method by Abbot Aeroset Autoanalyzer (USA) which is provided from Central Laboratory of Faculty of Medicine in Dicle University. The original kit of Abbot companies was used in the study. Determination of Hormonal Parameters For the hormonal parameters used in the study; T3, T4, TSH, LH, FSH, Testesterone, Estradiol, Prolactin, Cortisol, ACTH and growth hormone values were determined by electrochemiluminescence method using Roche Diagnostic E-170 (Germany) which is provided from Central Laboratory of Medicine Faculty of Dicle University. The original Roche kit was used in study.

Table 1. The statistical analyses of some biochemical parameters in rabbits before 900 MHz RF exposure. The values represent the mean ±S.D (Standard Deviation), Exp: Experimental.

Statistical Analysis Mean values and standard deviations were calculated, and statistical significance of the differences between exposure and control groups was evaluated. A computer program (SPSS 10.0, SPSS Inc., Chicago, IL, USA) were used for statistical analysis. Data were analyzed by MannWhitney U test which is a nonparametric test. All hypothesis tests used a criterion level of p = 0.05. Results As a result of experimental study done; before the exposure there was not any statistically significant differences found between the experimental and control group rabbits’ biochemical parameters such as Glucose, Creatinine, Na, K, Cl, Calcium, Phosphorus, Total Protein, Albumin, Globulin and Magnesium (Table 1). Also in analysis of biochemical parameters done after first month of RF exposure; there was not any statistically significant difference between the experimental and control groups (Table 2). Volume 2 ∙ Number ∙ 3 ∙ 2009

Table 2. The statistical analyses of some biochemical parameters in rabbits after first month of 900 MHz RF exposure. The values represent the mean ±S.D (Standard Deviation), Exp: Experimental. When evaluating the findings after 3 months RF exposure; again it was determined that there was not any significant difference regarding biochemical parameters (Table 3). In the present study it was attempted to be determine the changes on hormonal parameters Page 112


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THE EFFECT OF RF RADIATION Beyza KARADEDE et al

Table 3. The statistical analyses of some biochemical parameters in rabbits after third month of 900 MHz RF exposure. The values represent the Table 5. The statistical analyses of some hormonal mean ±S.D (Standard Deviation), Exp: Experimental. parameters in rabbits after first month of 900 MHz RF exposure. The values represent the mean ±S.D such as T3, T4, TSH, LH, FSH, Testosterone, (Standard Deviation), Exp: Experimental. Estradiol, Prolactin, Cortisol, ACTH and growth At the end of first month of exposure, it was hormone, after the RF exposure. In evaluation done before the exposure, it was found that there was not found that there was an increase in T4, any significant change in hormonal parameters of Testosterone, Estradiol, Cortisol and ACTH levels that is not statistically significant (Table 5). As for experimental and control group of rabbits (Table 4). analysis done on bloods taken from rabbits after 3 months exposure, there was an increase at T4, Cortisol and ACTH levels and only increasing in ACTH levels was stastistically significant (p<0.05)(Table 6). Values of average power density of RF radiation obtained from RF generators measured at different levels of rabbits’ bodies and they were as follows; at the level of rabbits’ nose 0.215 mW/cm2, at the level between two eyes 0.118 mW/cm2, at the level between two ears 0.114 mW/cm2, at the level of back 0.026 mW/cm2, at the level of rear leg 0.013 mW/cm2. The measured background RF level was equal to 0.0001 mW/cm2 when the RF generator is turned off. Discussion

Tablo 4. The statistical analyses of some hormonal parameters in rabbits before 900 MHz RF exposure. The values represent the mean ±S.D (Standard Deviation), Exp: Experimental. Volume 2 ∙ Number ∙ 3 ∙ 2009

In the present study, biochemical and hormonal effects of 900 MHz frequency RF radiation emitted from mobile phones were evaluated by determining the levels of some hormone and biochemical parameters. Also in this present study; to determine the cumulative effect of RF radiation depending on the time, changes at parameters studied were determined before Page 113


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Table 6. The statistical analyses of some hormonal parameters in rabbits after third month of 900 MHz RF exposure. The values represent the mean ±S.D (Standard Deviation), Exp: Experimental. exposure to RF, at first month of exposure and after third month of exposure. When biochemical data obtained was evaluated; although, there was increase and decrease in some variables after first and third month of exposure, it was determined that these changes were not significant. The hormones secreting from endocrine system are the important variables that regulate the metabolic activity. Therefore, it was investigated whether chronic exposure of 900 MHz RF radiation can affect to the important hormonal parameters such as T 3, T 4, TSH, LH, FSH, testosterone, estradiol, prolactin, cortisol, ACTH and growth hormone. In the present study, it was determined that; in first month of exposure; T 4, testosterone, estradiol, cortisol and ACTH levels were increased, but this increasing was not significant, however; after 3 months exposure, T 4, cortisol and ACTH levels were also increased, but from these changes, only an increasing in ACTH levels were significant. Endocrine system of mammals regulate the body functions to protect the homeostasis in cases of basal and stress situations (12). As RF radiation can be taken into account as a type of environmental stress, it is important to determine the effect of RF radiation emitted from mobile phones which are intensively used nowadays, on endocrine system and on the biochemical parameters, which are important for the functioning of biological systems. Different results are Volume 2 ∙ Number ∙ 3 ∙ 2009

THE EFFECT OF RF RADIATION Beyza KARADEDE et al

encountered in studies done regarding the effects of microwave and RF radiation, at different frequencies and different power densities, on hormones secreted by the endocrine system. Michaelson et al. found that GH level was decreased in rats which exposed to 2450 MHz frequency, 36 mW/cm2 power density microwave radiation for 60 minutes (13). These researchers observed that the decreasing in GH levels is associated with mean body temperature increasing up to 40.9 °C and making a peak. Lu et al. concluded that corticosterone and prolactin concentrations increased after the exposure of 2.45 GHz frequency amplitude modulated microwave radiation (14). Lotz and Podgorski measured the plasma cortisol, GH and T 4 levels in Rhesus monkey which were exposed to 1.29 GHz microwave radiation (15). These authors reported that GH and T 4 concentrations did not change but cortisol levels were increased during the exposure of 38 mW/cm2 power density RF radiation. Lu et al. concluded that the change in corticosterone, thyrotropin and GH levels can be correlated with power density or colon temperature of rats exposed to microwave for 1 hour (16). Some researchers observed an increase in plasma corticosterone levels in animals which were exposed to Radiofrequency Radiation (RFR) at the level of SAR (Specific Absorbtion Rate) that creates thermal stress (13,17,18-20). Liburdy suggested that thermal stress which formed as a result of whole body RFR exposure stimulated the release of adrenal steroids in blood (20.21). However, it has been reported that, adrenocortical inhibition occurs at SAR levels lower than 4 W/kg without a significant increase in colon temperature (19). Akdag et al. determined that 9450 MHz RF exposure can cause to significant decrease in GH and FSH levels and increase in estradiol levels of female rats (22). In the present study, at the first month of RF exposure, non significant increases in T4, testosterone, estradiol, cortisol and ACTH levels were determined but after third months of RF exposure T 4, cortisol and ACTH levels increased and only the increase in ACTH levels were significant. It has been proposed that low intensity microwave radiation can cause stress (4). Hormones like ACTH and cortisol are such hormones that generally associated with stress but GH and prolactin are hormones which is less associated with stress (4). In our study, the increasing occurred in cortisol and ACTH levels at the first and third months of the exposure can be due to stress induced by low-intensity RF radiation which is a stress factor. Page 114


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The results obtained in our study regarding the hormonal parameters are consistent with the data of the studies that investigate the effects of RF and microwave radiation on the neuroendocrine system (5,7,9,10). In the present study, there was not any changes in biochemical parameters which have important functions on the biological system, after RF exposure at both 1 month and 3 months. Therefore, it can be suggested that RF radiation obtained in our current experimental conditions are not enough to affect the biochemical parameters in rabbits. However, much longer-term researches that will be done on both experimental animals and human will help to answer the questions existing on this issue.

References

THE EFFECT OF RF RADIATION Beyza KARADEDE et al

15. Lotz WG and Podgorsky RP. Temperature and adrenocortical responses in rhesus monkeys exposed to microwaves. J.Appl.Physiol. 1982; 53; 1565-71. 16. Lu ST, Lebda N, Pettit S, Michaelson SM. Delineating acute neuroendocrine responses in microwave-exposed rats. J.Appl.Phsiol. 1980; 48: 927-32. 17. Magin RL, Shin-Tsu L and Michaelson SM. Microwave heating effect on the dog thyroid gland. IEEE. Trans. Biomed. Eng. 1977; BME24: 522-529. 18. Lotz WG, and Michaelson SM. Effects of hypophysectomy and dexamethasone on rat adrenal response to microwaves. J.Appl.Phsiol.1979; 47: 1284-88. 19. Lu ST, Lebda N, Pettit S, Michaelson SM. Microwave-induced temperature, corticosterone, and thyroptropin interrelationship. J.Appl.Physiol. 1981; 50: 399-405. 20. Liburdy RP. Radiofrequency radiation alters the immune system: Modification of T-and B-lymphocyte levels and cell-mediated immunocompetence by hyperthermic radiation. Radiat Res. 1979; 77: 34-46. 21. Liburdy RP. Radiofrequency radiation alters the immune system. II. Modulation of in vivo lymphocyte circulation. Radiat Res. 1980; 83: 6673. 22. Akdag MZ, Sert C, Kaya H, Dasdag S, and Celik MS. Effects of whole-body chronic microwave exposure on some hormones of variously treated rats. Biochemical Archives. 1999; 15: 345-350.

1. Vechhio GD, Giuliani A, Fernandez M, Mesirca P, Bersani F, Pinto R, Ardoino L, Lovisolo GA, Giardino L, and Calza L. Effect of radiofrequency electromagnetic field exposure on in vitro models of neurodegenerative disease. Biolectromagnetics. 2009; 30: 564-572. 2. IEEE, Standard for safety levels with respect to human exposure to radiofrequency electromagnetic fields, 3 kHz to 300 GHz. C95.1-1991. New York:IEEE, 1992 3. International Commission on Non-Ionizing Radiation Protection (ICNIRP), Health issues related to the use of hand-held radiotelephones and base transmitters. Health Phys.1996; 70: 587-593. 4. International Radiation Protection Assosiation (IRPA), Guidelines on limits of exposure to radiofrequency electromagnetic fields in the frequency range from 100 kHz to 300 GHz. Health Phys.1988; 54: 115123. 5. Seze de R, Fabbro-Peray P, and Miro L. GSM radiocellular telephones do not disturb the secretion of anteropituitary hormones in humans. Bioelctromagnetics. 1998; 19: 271-278. 6. Navakatikian MA, Tomashevskaya LA. Biological effects of electric and magnetic fields:Vol 1. Sources and Mechanisms. London, Academic Press, 1994. 7. Braune S, Riedel A, Schülte-Mönting J and Raczek J. Influence of a radiofrequency electromagnetic field on cardiovascular and hormonal parameters of the autonomic nervous system in healthy individuals. Radiat Res. 2002; 155: 352-356. 8. Rajkovic V, Matavulj M, Lukac T, Gledic B, Babic L, Lazetic B. Morphophysiological status of rat thyroid gland after subchronic exposure to low frequency electromagnetic field. Med Pregl 2001; 54: 119-127. 9. Mann K, Wagner P, Brunn G, Hassan F, Hiemke C, Röschke J. Effects of pulsed high-frequency electromagnetic fields on the neuroendocrine system, Neuroendocrinology. 1998; 67: 139-144. 10. Stagg RB, Hawel LH, Pastorian K, Cain C, Adey WR, and Byus CV. Effect of immobilization and concurrent exposure to a pulse-modulated microwave field on core body temperature, plasma ACTH and corticosteroid, and brain ornithine decarboxylase, Fos and Jun mRNA, Radiat Res. 2001; 155: 584-592. 11. Radon K, Parera D, Rose DM, Jung D, and Vollrath L. No Effects of pulsed radio frequency electromagnetic fields on melatonin, cortisol, and selected markers of the immune system in man. Bioelectromagnetics. 2001; 22; 280-287. 12. Abhold RH, Ortner MJ, Galvin MJ and McRee DI. Studies on acute in vivo exposure of rats to 2450 MHz microwave radiation. II. Effects on thyroid and adrenal axes hormones. Radiat Res. 1981; 88: 448-455. 13. Michaelson SM, Houk WM, Lebda NJA, Lu ST, Magin RL. Biochemical and neuroendocrine aspects of exposure to microwaves. Ann. N.Y. Acad. Sci. 1975; 247: 21-45. 14. Lu ST, Lebda N, Lu SJ, Pettit S, Michaelson SM. Effects of microwaves on three different strains of rats. Radiat Res. 1987; 110: 173-91.

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