In order to perform the protein analysis using the paraffin sections previous fixed with formalin, we applied the ImmunoMemBlot (IMB) method1) to detect the epitopes of target proteins with specific antibodies. In this study the protein extracts were obtained from the paraffin sections of each representative case of ameloblastoma, adenomatoid odontogenic tumor (AOT), and normal gingiva, and more a protein extract from fresh tissue of ameloblastoma was also compared to evaluate the IMB results used with 24 different antibodies. First of all, in the comparison between the paraffin section extract and fresh tissue extract of ameloblastoma, the latter consistently showed more positive IMB reaction than the former. Meanwhile, the paraffin section extract of ameloblastoma was more comparable with that of normal gingival, disclosing that most of proliferating genes, oncogenes, and apoptosis related genes, i.e., PCNA, CDK4, c-erbB2, CEA, p53, Bax, Bad, FLIP, FAS, Bcl-2, p21, N-ras, MMP-2, MMP-9, caspase-3, -8, -9, were highly expressed in ameloblastoma, but EGFR, HGF, and VEGF were similarly expressed both in the ameloblastoma and in normal human gingiva. On the other hand, the comparison between ameloblastoma and AOT both in the immunohistochemistry and IMB using their paraffin section extracts clearly demonstrated that the ameloblastoma showed more expression of proliferating genes and oncogenes while the AOT showed more expression of apoptosis related genes, i.e., Bax, Bad, FLIP, and caspase-9. Taken together, these data suggest that the IMB can be used for the primary screening of quantitative protein analysis using the paraffin section extract, and that the IMB results could be evaluated in conjunction with the immunohistochemical observation.
Hemangiomas are different from true vascular malformations in thei l‘ pathogenesis and cl inical prognosis. There are sti ll no standardized antibodies to distinguish hemangioma and vascular malformation apparently. We compared juvenile hemangioma and vascular malformation with immunohistochemjstry using va ri OllS antibodies, i.e. , ANG, bFGF, VEGF. EGFR, vWF. PCNA. p53. maspin, and TNF- . A very st rong positive expression of ANG and vWF was observed mainly in the vascular endothelial cells of juvenile hemangioma. VEGF s howed st rong positive reaction in the juveni le hemangioma, but p53 showed no positive reaction. Ancl a strong positive reaction of ANG was observed in the vascular endothelial walls of vascular malformation. p53 was frequently positive in the lining endothel ial cells in the vascular malformatJOn Using a ntiboclies such as VEG F'. ANG. vWF which a re related to the proliferation and matllrity of the vessel components. and p53 antibodies in order to confirm between juvenile hemangioma and vascular malformation would be helpful
In order to obtain novel genes related to the human craniofacial development, molecular cloning and sequencing, and in situ hybridization using craniofacial tissue sections were performed and followed by protein structure simulation. Totally 231 clones were obtained from the subtracted craniofacial tissue cDNA library of human embryo. Random cloning using the non-redundant clones from the craniofacial tissue of human embryo was done and obtained 398 clones from the premade human chondrocyte cDNA library. Their partial sequence data showed that 214 clones of subtracted cDNA library of craniofacial tissue were still non-redundant in Genebank search. And 20 clones among 498 clones of premade chondrocyte cDNA library were known to be undefined genes. Through in situ hybridization screening in the craniofacial tissue sections of 10 weeks old human embryo 36 clones were found to be positive in specific tissues. Depending on the cell types of sirnilar developmental origin, the positive reactions could be divided into five groups. Among the 20 clones of undefined genes from human chondrocyte cDNA library, 7 clones showed characteristic positive reaction in human cartilage tissue by in situ hybridization. From the simulated protein structure, motif analysis and in situ hybridization studies for the 7 undefined clones, Ch89, Ch96, Ch129, Ch285 clones may function in the outer space of the cell constituting a part of matrix protein complex, and Ch276 as a transmembrane protein which might partic ipate in matrix calcification around chondrocytes. Ch153 is a kind of antirnicrobial protein also acting as an inflammation mediator, and Ch334 clone is a zinc finger protein, of which expression increases in human adult tissues We presume these novel genes from human chondrocytes may provide a new path of chondrocyte development and functions of human craniofacial tissues
The fragile X mental retardation (FMR) syndrome is the largest source of inherited mental retardation. The syndrome is usually caused by the transcriptional silencing of fragile X mental retardation gene (FMR-1). An 18 years old male wascompla띠ing of multiple toαh missing and abnormal facial profùe, of which signs were matemally dominant in his family. In the C)π。gene디c analysis the pa디ent and his parents did not show any discontinuity in the long arm end of X chromosome, but in the PCR produαs targeω19 the CGG repeat sequence in the 5' untranslated region of FMR gene both the patient'’ s and his mother' s gDNAs produced a normal and an extra bands, sized about 400 and 800 bps, respectively, while the his father' s gDNA produced only one normal band, sized about 400 bps. 까1US , we suppose that the pa디.ent has heterozygotic alleles of FMR gene inherited matemally, and that the patient s FMR gene was in a premutated state relevant to the dentofacial abnormalities.
The temporomandibular joint (TMJ) showed characteristic anatomy and functions contrast to the other joints. In particular, children or young adolescent with the progressive temporomandibular disorder (TMD) showed remarkable changes in their mandibular growth and occlusion. The purpose of this study was to elucidate the functional and biomechanical causes for the maxillofacial deformities of TMD. A typocranial experiment was performed using a plastic skull model by simulating the mandibular movement after condylectomy. The directional forces of masticatory and suprahyoid muscles were reproduced on the typocranial skull model by elastics connecting between mandible and adjacent bony structures. Resultantly, the mandible of unilaterally or bilaterally condylectomized mandible was rotated downward and backward by the activation of masticatory and suprahyoid muscles. When unilaterally or bilaterally condylectomized mandible was activated by masticatory, suprahyoid and perioral muscles, the mandible was less rotated downward and backward than the cases activated by masticatory and suprahyoid muscles only. In the typocranial experiment after unilateral condylectomy the mandible was rapidly deviated toward the affected side, but this deviation was much reduced when the mandible was intensely supported by perioral muscles. By the compensatory response against the overload of TMJ due to the mandible body resection the typocranial experiment also produced the deepening of antegonial notch by the increased force of mylohyoid and perioral muscles. These findings suggest that after condylectomy the posterior fibers of mylohyoid and perioral muscles become hypertrophic and induce the characteristic mandibular deformities, i.e., deepening of antegonial notch and bowing of posterior ramus, which are similar to the mandibular changes in the progressive TMD.
Osteoarthrosis in TMJ is known to accompany characteristic features of the craniofacial malformation and malocclusion. In order to explain the mechanism that brings about the typical mandibular changes relating to the frequent clinical symptom, i.e., the limited mouth opening in TMJ patients. The present study investigated the mandibular morphology of the rabbit, also intentional TMJ osteoarthrosis was performed on the TMJ of the rabbits and their morphological changes were observed. A total of 12 New Zealand house rabbits weighing an average of 3 kgwere used and all left side joints were surgically meniscectomized and right side joints were used as controls. The rabbits were sacrificed in one week, two weeks, one month, three months and 5 months after the menisectomy, and the removed TMJs were examined by histological and radiographical methods and analyzed statistically. It was found that the mandibular morphology caused by osteoarthritis in the TMJ was characterized by the decrease in size of the ascending ramus, posterior ramus bowing, deep antegonial notch and thickening of the cortex in inferior and posterior borders of the mandible, that the similar features were also observed in the rabbit mandible. When the menisectomy was performed in the unilateral side of rabbit TMJ, the mandibular changes including the increased depth of antegonial notch and posterior ramus bowing occurred only on the affected side. These data suggested that the limitation of the mandibular movement and altered masticatory pattern by meniscectomy affects the structural remodeling of the mandible and TMJ likely the TMJ osteoarthrosis
As pulp calcification occurs at least fifty percent of total teeth, the focal calcification in pulp chamber usually appears in all age groups. However, the pulp calcification is one of the important pathologic changes affecting the pulp vitality. In order to elucidate the mechanism of pulp calcification during the retrogressive degeneration of pulp tissue we performed an immunohistochemical study for proteases (MMP-3, MMP-10, and cathepsin-G), antiproteases (TIMP-1, α1- AT) and proteins involving tissue protection (TGase-2 and HSP-70). In the normal pulp tissue MMP-3 and MMP-10 were weakly expressed, but cathepsin-G and TIMP-1 were rarely expressed. Around the calcifying tissue of MMP-3, MMP- 10, and α1-AT were predominant, but TIMP-1 and cathepsin-G were sparsely expressed. On the other hands, TGase-2 and HSP-70 were condensed in the proximal fibrous tissue. These data suggest that the pulp calcification is related to retrogressive pulp degeneration, which could be resulted in the incomplete digestion of the degenerated stromal tissue by different proteases. We presume that the aberrant protease digestion of chronic pulpal pathosis, i.e., sclerotic fibrosis, chronic pulp degeneration, etc., may enhance the dystrophic calcification in dental pulp.
The primary growth center (MdPGC) of human fetal mandible was conspicuously distinguished in the soft X-ray view of fetal mandibles.1) As the peripheral adaptive growth of mandible advanced during the postnatal period, the MdPGC became overshadowed by condensed cortical bone. However, in the well-processed radiograms of adult mandible a condensed radiopaque image, measuring 0.5-1.0 cm in diameter, can be observed below the apex of first premolar. In this study we aimed to trace a sclerotic sequela of mandibular primary growth center during postnatal period. Panoramic radiograms of two hundreds adults and soft X-ray views of thirty dry mandible were analyzed by statistical methods. The adult MdPGC was clearly distinguishable from the mental foramen. The area of MdPGC was seldom changed in the older persons, even in the edentulous mandibles. Additionally, the benign lesions of odontogenic cysts and tumors hardly destroyed the original structure of MdPGC, while the malignant tumors of squamous cell carcinoma and metastatic cancer rapidly destroyed and resolved the radiopaque area of the MdPGC.
Background : Acanthopanax sessiliflorus (Rupr. et Maxim) Seem, belonging to the Araliaceae family, is widely distributed in Korea, China, and Japan. The plants belonging to Acanthopanax species are traditionally used in Korea as anti-rheumatoid arthritis, anti-inflammatory and anti-diabetic drugs and are recognized to have ginseng-like activities. A simple and sensitive high-performance liquid chromatographic (HPLC) method was developed and validated for independent analysis of major compounds and chlorogenic acid in A. sessiliflorus fruits. Chlorogenic acid was reported that prevent cancer and cardiovascular disease in vivo. Also, it has antioxidant effect in vitro test. In the previous experiment, chlorogenic acid were found in A. sessiliflorus fruits. This study was performed to identification of the major compounds and investigate the method validation for the determination of chlorogenic acid in A. sessiliflorus fruits. Methods and Results : Three major compounds were recorded on a Varian Unity Inova AS-400 FT-NMR spectrometer and analyzed by the new HPLC analysis method. HPLC analysis was carried out using an Waters e2695 and PDA detector. The new analyasis method was validated by the measurement of intra-day, inter-day precision, accuracy, limit of detection (LOD, S/N=3), and limit of quantification (LOQ, S/N=10) of chlorogenic acid. The results showed that the correlation coefficient (R2) for the calibration curves of chlorogenic acid was 0.997 in terms of linearity. The limit of detection (LOD) and limit of quantification (LOQ) were 0.565 ㎍/ml and 2.88 ㎍/ml, respectively. There was no interfering peak observed each other and HPLC system was suitable for analysis showing goodness of peak and high precision. Conclusion : This method is suitable to detect and quantify major compounds in A. sessiliflorus fruits. Furthermore, the result will be applied to establish chlorogenic acid as an standard compound for A. sessiliflorus fruits.
In recent localized heavy rainfalls have been arising from abnormal climate change. People are concerning about damages with increasing the frequency of flooding. Therefore, we need to understand river hydraulic characteristics and management to reduce damage from flooding. To study hydraulic characterization of Sincheon experimental catchment HEC-RAS (Hydrologic Engineering Center River Analysis System) model which provided by U.S Army Corps of Engineers (USACE) was applied. This study analyzed and compared water level the frequency flood for 100 years and 200 years by clark unit Hydrography. The change of the water level of Daejeon bridge, Sincheon bridge and Singi bridge showed increased for all conditions. The flow rate for the Daejeon bridge and the Sincheon bridge showed an increase, but the Sinki bridge showed a decreasing flow rate overally, except for 1hour-100 years. The verification result showed that the model was able to simulate the water level with 0.4709 coefficient of determination and error ration ranging from 1 to 3%.
Acinetobacter baumannii is usually considered an opportunistic pathogen that it responsible for a variety of nosocomial infections, such as, pneumonia, tracheobronchitis, meningitis, endocarditis, peritonitis, skin and soft tissue infections, and urinary tract infections. However, over the years, the organism has developed substantial antimicrobial resistance, and thus, the management of infections has become more difficult. The less common infective endocarditis is one of the more serious consequences of nosocomial bacteremia. Here, we report the successful treatment of the first case of multidrug-resistant A. baumannii endocarditis in a 33-year-old patient.