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Gene expression and protein localization of amelogenin in tooth germs and odontogenic tumors
P23-3 Pathology (cyst and others) known to possess antioxidant like activity which may inhibit the production of free radicals. Strenuous exercise can act as a source of external oxidative stress and is also known to affect the composition and functional activity of saliva. Few studies have been carried out to study the changes in antioxidant enzymes in salivary glands. The present treadmill exercise study, was undertaken in adult ICR female mice, maintained on a chow diet fed ad libitum and with or without exercise for 3 months, to measure both transforming growth factor beta-1 (TGF]31) expression by RT-PCR and antioxidant enzyme levels in the salivary glands. Weekly body weights were monitored closely and at the end of study both sedentary and exercised mice were killed to obtain salivary glands to measure TBA-reactive products and various antioxidant enzymes. Mice were exercised 5 days/week, ran approximately 1 km a day (1 hour). Exercised animals body weights were to be found reduced significantly (16.5%, P<0.05) as compared to sedentary mice at 3 months. Significantly lowered (30.5%) levels of TBA-reactive products in salivary glands were observed in exercised mice as compared to sedentary mice. Antioxidant enzyme levels such as catalase, superoxide dismutase and glutathione-S-transferase activities were significantly (P<0.05) increased, whereas, TGF[31 m R N A expression in salivary glands was found to be decreased in exercised mice. These results indicate that increased antioxidant enzyme activities in exercised mice may be due to the decreased proinflammatory TGF[31 expression which may thereby prevent exercise induced oxidative damage in the salivary gland tissues.
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cultured specimen as early as 7 hours after infection, and maintained even after 8 days in culture. These results illustrate model systems that can be used to characterize cranial neural crest cell migration and differentiation employing the mouse E8 organ culture system and H.H. stage 9 tol 1 chick rhombomere explant culture system coupled with ectopic gene expression by adenovirus delivery. This study was supported by N I H Grant Z01-AR41114.
10. Gene Expression and Protein Localization of Amelogenin in Tooth Germs and Odontogenic Tumors
Nagai, N.1, Nakano, K. 1, Gui-Ru, L. 1, Chigono, yz, Takagi, T.3, Hong-ln, S. 4 1Department of Oral Pathology, Okayama University Dental School 2Department of Oral Surgery, Teikyo University School of Medicine, 3Department of Biochemistry, Faculty of Dentistry, Tokyo Medical and Dental University, Japan, 4Department of Oral Pathology, School of Dentistry, Kyongpook National University, Korea In situ hybridization A Digoxigenin labeled R N A probe was hybridized with tissues at 50~ for 16 hours. After the excess amelogenin probe was washed off, Dig Nucleic Detection Kit was used to visualize the hybridized probe.
Rat incisors The amelogenin gene was expressed in the preameloblasts and secretory ameloblasts of the inner enamel epithelium from the time when predentin appeared. 9. Adenovirus-Mediated Ectopic Gene Expression Using Chicken and Mouse Craniofacial Explant Cultures
Takahashi, If., Tanaka, 0., Nuckolls, G. H., Shum, L., Slavkin, H. C.
Craniofacial Developmental Section, NIAMS, NIH, Bethesda, Maryland, U.S.A. Of all congenital malformations, 25% affects the craniofacial region. In order to understand the basic mechanisms of craniofacial dysmorphogenesis, we study the migration of cranial neural crest cells emigrating from rhombomeres and their differentiation, in branchial arches. Tissue wedge including rhombomeres 1 and 2 of the hindbrain, and developing first branchial arch region were microdissected from E8 (3-6 somite stages) Swiss Webster mouse embryos and explanted into organ culture in BGJb medium supplemented with 50% Ic rat serum and l m M pyruvate. Similary, individual rhombomeres were microdisected out from H.H. stage 9 t o l l chicken embryo. The isolated segments were incubated on the fibronectin-coated dish in F12 medium supplemented with 10% FBS. We demonstrated that ectopic gene expression in such murine and avian rhombomere explant cultures was achieved by exposing explants to infection by adenovirus carrying lacZ or green fluorescent protein (GFP) construct. Positive signals were detected in
Ameloblastomas The tumor cells of ameloblastomas were so well differentiated as to express the amelogenin gene. However, the distribution patterns of amelogenin m R N A varied with histological types. Amelogenin protein was occasionally detected (2 of 10 cases). Adenomatoid Odontogenic Tumors (AOT) The expression patterns of amelogenin m R N A varied according to the morphological differences of tumor cells. Amelogenin protein was detected in the tubular cavities, around tumor cells and in the calcified matrix. AOT was composed of different cells showing various differentiated functions, from cells expressing amelogenin m R N A and protein to cells negative in both m R N A and protein expression. The tumor cells expressing neither amelogenin m R N A nor amelogenin protein were similar to those of the reduced dental epithelium in tooth germ.
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cultured specimen as early as 7 hours after infection, and maintained even after 8 days in culture. These results illustrate model systems that can be used to characterize cranial neural crest cell migration and differentiation employing the mouse E8 organ culture system and H.H. stage 9 tol 1 chick rhombomere explant culture system coupled with ectopic gene expression by adenovirus delivery. This study was supported by N I H Grant Z01-AR41114.
10. Gene Expression and Protein Localization of Amelogenin in Tooth Germs and Odontogenic Tumors
Nagai, N.1, Nakano, K. 1, Gui-Ru, L. 1, Chigono, yz, Takagi, T.3, Hong-ln, S. 4 1Department of Oral Pathology, Okayama University Dental School 2Department of Oral Surgery, Teikyo University School of Medicine, 3Department of Biochemistry, Faculty of Dentistry, Tokyo Medical and Dental University, Japan, 4Department of Oral Pathology, School of Dentistry, Kyongpook National University, Korea In situ hybridization A Digoxigenin labeled R N A probe was hybridized with tissues at 50~ for 16 hours. After the excess amelogenin probe was washed off, Dig Nucleic Detection Kit was used to visualize the hybridized probe.
Rat incisors The amelogenin gene was expressed in the preameloblasts and secretory ameloblasts of the inner enamel epithelium from the time when predentin appeared. 9. Adenovirus-Mediated Ectopic Gene Expression Using Chicken and Mouse Craniofacial Explant Cultures
Takahashi, If., Tanaka, 0., Nuckolls, G. H., Shum, L., Slavkin, H. C.
Craniofacial Developmental Section, NIAMS, NIH, Bethesda, Maryland, U.S.A. Of all congenital malformations, 25% affects the craniofacial region. In order to understand the basic mechanisms of craniofacial dysmorphogenesis, we study the migration of cranial neural crest cells emigrating from rhombomeres and their differentiation, in branchial arches. Tissue wedge including rhombomeres 1 and 2 of the hindbrain, and developing first branchial arch region were microdissected from E8 (3-6 somite stages) Swiss Webster mouse embryos and explanted into organ culture in BGJb medium supplemented with 50% Ic rat serum and l m M pyruvate. Similary, individual rhombomeres were microdisected out from H.H. stage 9 t o l l chicken embryo. The isolated segments were incubated on the fibronectin-coated dish in F12 medium supplemented with 10% FBS. We demonstrated that ectopic gene expression in such murine and avian rhombomere explant cultures was achieved by exposing explants to infection by adenovirus carrying lacZ or green fluorescent protein (GFP) construct. Positive signals were detected in
Ameloblastomas The tumor cells of ameloblastomas were so well differentiated as to express the amelogenin gene. However, the distribution patterns of amelogenin m R N A varied with histological types. Amelogenin protein was occasionally detected (2 of 10 cases). Adenomatoid Odontogenic Tumors (AOT) The expression patterns of amelogenin m R N A varied according to the morphological differences of tumor cells. Amelogenin protein was detected in the tubular cavities, around tumor cells and in the calcified matrix. AOT was composed of different cells showing various differentiated functions, from cells expressing amelogenin m R N A and protein to cells negative in both m R N A and protein expression. The tumor cells expressing neither amelogenin m R N A nor amelogenin protein were similar to those of the reduced dental epithelium in tooth germ.