Diffuse large B-cell lymphoma (DLBCL) is the most common non-Hodgkin's lymphoma, and usually showed painless neck swelling, fever, sweat, and weight loss. Although about 5% of all lymphomas appeared in the oral area, the primary maxillofacial lymphomas were rare and sometimes clinically tended to be misdiagnosed such as chronic periodontitis, osteomyelitits, etc. This study demonstrated three cases of primary DLBCL mimicking localized osteomyelitis at mandible or maxilla. A series of histological and immunohistochemical examination using different biomarkers of lymphoreticular cells were performed to characterize the neoplastic cells of DLBCL. The first case occurred in a 45 years old male exhibiting mandibular osteomyelitis and neck swelling. The second case simply showed a gingival swelling at left upper premolar area in a 55 years old male. And the third case is from an 84 years old female who felt numbness at left lower lip and had severe periodontitis involving regional alveolar bone resorption. All of three cases had experienced no systemic manifestation of lymphoreticular malignancy before the diagnosis of oral lymphoma. Immunostainings of CD3, CD20, TNFα, BCL-2, Ki-67, PCNA, and c-Myc were strongly positive in these tumor cells, while those of p53 and CD31 were slightly positive, and CD56 immunoreaction was negative. These three cases were diagnosed as DLBCL and referred to the hemato-oncology unit for treatment. Therefore, every chronic granulomatous periodontal lesion hardly cured by simple medical treatment should be carefully explored through pathological examination, and it was presumed that DLBCL is closely related to the chronic inflammatory periodontal lesions recruiting mucosa-associated lymphoid cells in older patients. It was also suggested that DLBCL be differentially diagnosed from T-cell lymphoma, Burkitt’s lymphoma, and Hodgkin’s disease, etc. with immunohistochemical determination of tumor cell subtypes as soon as possible in order to be treated with appropriate therapy.

A 31 years old female had been suffered from a bony swelling in right premolar region of the mandible for 12 years, recently grown rapidly. A fistula tract developed on the right anterior mandibular border, but the lesion was relatively asymptomatic. In the radiological examination, the tumor mass was irregularly mixed with radiolucent and radiopaque areas, forming multiple cystic spaces. Under the diagnosis of calcifying odontogenic cyst, the mandibular mass was resected and examined pathologically. After decalcification, the dissected tumor mass showed multiple small cystic spaces and calcifying fibrous tissue, mimicking calcifying odontogenic cyst or ameloblastoma. Histological observation showed many calcifying cementoid materials and ossifying trabeculae. The cystic spaces were turned out to be dilated vascular channels lined by endothelial cells, containing plasma fluid. However, the main lesion was diagnosed as cemento-ossifying fibroma (COF), and the atypical vascular channels were greatly dilated and gradually expanded the whole tumor mass. The present COF was examined through immunohistochemical (IHC) array, and investigated for tumor cell characteristics, exhibiting abnormal ossification and aneurysmal cystic changes. IHC array disclosed that the tumor cells grew progressively in the lack of apoptosis, and that they showed lower expression of RUNX2 than BMP-2, RANKL, and OPG, and increases of protein expression in HIF-1α, VEGF-A, and CMG2. These data suggested that the reduced expression of RUNX2, osteoblast differentiation factor, be relevant to abnormal ossification of COF, and that the consistent expressions of angiogenesis factors be relevant to de novo angiogenesis in COF, subsequently resulted in aneurysmal cystic changes.

A 57 years old female complained of severe pain on the right temporomandibular joint (TMJ) area. Her right condyle had been partly resected under surgical operation 13 years ago due to condyle hypertrophy, thereafter she felt dull pain on TMJ area and recently the lesion became severely swelled and painful leading to cancer phobia. The present radiological views showed slightly enlarged and sclerosed condyle with increased radiopacity, but its articular sliding function was almost disable during mouth opening. The patient’s TMJ lesion was carefully managed with conservative physiotherapy and pain treatment. The microsection of condyle head obtained from the previous operation was re-evaluated histologically, and it was finally diagnosed as osteochondrosis dissecans (OCD), exhibiting hyperplastic proliferation of cartilage in condyle head and marked vascular dilatation in epiphyseal zone. This abnormal cartilage tissue was distinguishable from normal cartilage tissue found in the peripheral cartilaginous cap of the same microsection. The involved cartilage cap showed thick hypertrophic chondrocyte zone with horizontal and vertical clefts accompanying diffuse hyaline degeneration. The superficial fibrous zone of cartilage cap was thickened and frequently peeled off, while lower hypertrophic zone of cartilage cap was highly cellular and proliferative. Consequently, the endochondral ossification became aberrant and resulted pre-mature apoptosis of many hypertrophic chondrocytes, followed by diffuse and mild inflammatory reaction in the underlying marrow tissue. Therefore, it was suggested that this hypertrophic condyle lesion, OCD, be differentiated depending on radiological and histological features from ordinary condyle hyperplasia, osteochondroma, and osteoarthritis, and that the pathological confirmation of OCD may provide a reliable modality for dental and medical treatment of chronic and painful TMJ lesion.

Styela clava (SC), a sea squirt has a biphasic life cycle, showed the animal phase during larval life and the vegetable phase during adult life. The adult SC becomes fixed on hard surface and covered with thick cellulose membrane to protect itself from various physical and chemical damages in marine environment. Especially, adult SC forms a huge cystic space as a portion of digestive tract, filled with seawater containing various planktons. In the present study, it is hypothesized that SC can produce secretory materials, like saliva in mammals, to sterilize, to neutralize, and to digest the cystic fluid. First of all, the cyst fluid was examined for the antimicrobial property by bacteria killing assay (BKA) using E. coli, and it resulted that the cyst fluid showed relatively weak bactericidal effect. The cyst fluid was also examined for antioxidant property by DPPH (1,1-diphenyl-2-picrylhydrazyl) and ABTS (2,2-azinobis-3-ethybenzthiazdine-6-sulphonic acid) assay, and resulted that the cyst fluid showed strong antioxidant effect comparable to ascorbic acid. The antioxidant materials of cyst fluid were found both in cationic and anionic groups separated by ion exchange column using SP and Q beads, and they were also heat resistant. The strong antioxidant property seems to be originated from the abundant carotenoid materials in the cyst fluid of SC. However, the strong antioxidant materials of the cyst fluid may play a role for the essential antioxidant in the digestive tract of SC. Furthermore, the cyst fluid of SC was examined using SNU-1 cancer cells for its anticancer property. The flow cytometry analysis showed that both of cationic and anionic materials purified from the cyst fluid showed strong anticancer effect on SNU-1 cells. Taken together, the cyst fluid of SC showed weak antimicrobial, but strong antioxidant property and marked anticancer properties in vitro experiment. Therefore, it is presumed that SCs can neutralize the various free radicals of seawater and also protect themselves from carcinogenic impacts in aggressive marine environment by various bioactve substances of their cyst fluid.

Coffee (Coffea spp.) is one of the most important agricultural commodities, being widely consumed in the world. Various beneficial health effects of coffee have been extensively investigated, but data on habitual coffee consumption and its bio-physiological effect have not been clearly explained as well as it is not proved the cause and effect between drinking coffee and its bio-physiological reactions. We made the dialyzed coffee extract (DCE), which is absorbable through gastrointestinal tract, in order to elucidate the cellular effect of whole small coffee molecules. RAW 264.7 cells, a murine macrophage lineage, were directly treated with DCE, i.e., DCE-2.5 (equivalent to 2.5 cups of coffee a day), DCE-5, and DCE-10, for 12 hours, and their protein extracts were examined by immunoprecipitation high performance liquid chromatography (IP-HPLC). RAW 264.7 cells differently expressed the inflammation-related proteins depending on the doses of DCE. RAW 264.7 cells treated with DCE showed marked increase of cathepsin C, cathepsin G, CD20, CD28, CD31, CD68, indicating the activation of innate immunity. Particularly, the macrophage biomarkers, cathepsin G, cathepsin C, CD31, and CD68 were markedly increased after DCE-5 and DCE-10 treatments, and the lymphocyte biomarkers, CD20 and CD28 were consistently increased and became marked after DCE-10 treatment. On the other hand, RAW 264.7 cells treated with DCE showed consistent increase of IL-10, an anti-inflammatory factor, but gradual decreases of different pro-inflammatory proteins including TNFα, COX-2, lysozyme, MMP-2, and MMP-3. In particular, the cellular signaling of inflammation was gradually mitigated by the reduction of TNFα, COX-2, IL-12, and M-CSF, and also the matrix inflammatory reaction was reduced by marked deceases of MMP-2, MMP-3, and lysozyme. These anti-inflammatory expressions were consistently found until DCE-10 treatment. Therefore, it is presumed that DCE may have dynamic effects of innate immunity activation and pro-inflammation suppression on RAW264.7 cells simultaneously. These effects were consistently found in the highest dose of coffee, DCE-10 (equivalent to 10 cups of coffee a day in man), that might imply the small coffee molecules were accumulated in RAW 264.7 cells after DCE-10 treatment and produce synergistic cytokine effects for innate immunity activation and anti-inflammatory reaction concurrently.

Coffee is one of the most familiar beverages to modern human adults, but its bio-physiological effect has not been clearly elucidated. It was known that more than one thousand chemicals were included in the ordinary coffee extract. Among them, the caffein and chlorogenic acid (caffeoylquinic acids) are most abundant and have been investigated by many authors so far. In order to know the real cellular effect of whole coffee extract elements, the dialyzed coffee extract (DCE)1) was made to get coffee elements less than 1000 Da molecular weight, which are freely absorable through gastrointestinal tract. It was directly treated in the culture of RAW 264.7 cells, a murine macrophage lineage. RAW 264.7 cells were treated with DCE equivalent to 2.5 cups of coffee (DCE-2.5), DCE-5, and DCE-10 for 12 hours, and their protein extracts were examined by histological observation and immunoprecipitation high performance liquid chromatography (IP-HPLC). RAW 264.7 cells differently expressed the proliferation-related proteins depending on the dose of DCE. DCE-2.5 and DCE-5 enhanced the cellular growth of RAW 264.7 cells by increasing the expression of β-actin, PCNA, Ki-67, MPM2, MAX, cMyc, E2F-1, and Rb-1, and by decreasing the expression of MAD and p21. These proliferation-related proteins were rarely affected by DCE-10. DCE-2.5 and DCE-5 induced the cellular proliferation of RAW 264.7 cells by the signaling of E2F-1 and cMyc, respectively, but these cellular effects almost disappeared in DCE-10. Therefore, it was presumed that the low dose of coffee, DCE-2.5 and DCE-5 might be effective for the proliferation of murine macrophages, RAW264.7 cells, contrast to the high dose of coffee, DCE-10. It was also suggested that the low dose of DCE-2.5 and DCE-5 be helpful to increase the innate immunity in vivo by increasing the cell number of macrophages in contrast to the high dose of DCE-10.

With the multiple practices of bone graft using different artificial bone regenerative substitutes, the bone graft procedures have been widely performed to increase the bony stabilization of dental implant. Xenogenic bone graft materials have been well developed because of their good biocompatibility and abundant source of bone materials. The present study demonstrated the histological findings from excellent bony remodeling in xenogenic bone graft biopsies compared to those findings in autogenous bone graft. For the graft bone biopsies which were usually done in 5-9 months after graft bone insertion, five types of histological grades including excellent, favorable, partial, degenerative, and poor bony remodeling could be assessed to give prognostic information for dental implant. However, recently the xenograft bone materials have been much improved and produced strong osteogenic effect. Among 239 cases of trephine bur-supported core bone biopsy the excellent bony remodeling was found in 20 cases (13.1%) out of 153 xenogenic bone grafts and in 13 cases (43.3%) out of 30 autogenous bone grafts. They produced abundant new bones on the surface of the graft bones in 5–9 months, and the graft bones were partly resorbed and also surrounded by the repetitive deposition of new bone. The osteophytic new bones showed strong birefringence under polarizing microscope, and were gradually elongated and anastomosed with each other to form trabecular bony networks which became proper stress-baring structures for dental implant. Their marrow stromal tissues were composed of loose connective tissue which was well vascularized but rarely infiltrated with inflammatory cells. The present study compared the histological features of excellent bony remodeling between xenogenic and autogenous bone grafts. Although the ratio of excellent bony remodeling in xenogenic bone graft was still low, 13.1%, the recent advance of xenogeic bone products was remarkable in biological aspect and almost comparable to the autogenous bones. Therefore, it was suggested that the xenogenic bone graft will be applicable to the bone regeneration procedures for dental implant with beneficial output in the near future.

In order to perform the biological investigation of coffee extract containing different molecules, it would be necessary to develop in vitro experimental system rather than animal experiment. Although the animal experiment treated via oral intake or intravenous injection may disclose the whole systemic effect, the in vitro cell culture experiment would be more convenient to analyze direct cellular effect of coffee extract than animal experiment. Therefore, this study was aimed to develop a dialysis method for the crude coffee extract to perform the biological investigation using murine macrophage cell line, RAW 264.7. First of all, the RAW 264.7 cells treated with dialyzed coffee extract were observed, and subsequently their protein extracts were analyzed by gel filtration chromatography, thin layer chromatography, and immunoprecipitation high performance liquid chromatography (IP-HPLC). Resultantly, it was found that the low dose (20μg/mL) of dialyzed coffee extract, about 5 cups of ordinary coffee drinking for human adult, enhanced the growth of RAW 264.7 cells by increased expression of β-actin and Ki-67, and also induced the anti-inflammatory effect by decreased expression of NFkB, TNFα, and LC3 contrast to the high dose (40μg/mL) of dialyzed coffee extract. The low dose of dialyzed coffee extract produced almost no harmful effect on RAW cell culture for 12 hours, rather than it produced stimulatory effect on RAW cells by increasing the cell number and enhancing the protein expression of β-actin, Ki-67. Therefore, it was thought that the low dose of dialyzed coffee extract is applicable to cell culture experiment without difficult purification procedures of coffee elements. In addition, as the contrast cellular effect between the low and high dose of coffee extract was found in this study, it was also presumed that the low dose of coffee extract may play an important role in the inflammatory reaction of murine macrophages.

The native human saliva obtained through the centrifugation of whole saliva showed characteristic salivary protein complex (SPC) peaks in gel filtration high performance liquid chromatography (HPLC) using Superose 12 column1,2). In the previous study the SPC peaks in chromatography were explored to know their composition and functions by different detection methods, but still the nature of SPCs was not clearly elucidated so far. In this study the SPC peaks were examined by direct antibody interaction in order to target different antimicrobial and protective proteins distributed in the SPCs via gel filtration HPLC. As the SPC peak shape and migration speed can be changed by antibody binding to specific proteins of SPC, it was found that mucin1 is evenly distribution in all SPCs, while PRPs are more abundant in the late dominant SPC than the early dominant SPC and also in the intermediated SPCs. Most of antimicrobial proteins including lysozyme, LL-37, lactoferrin, β-defensin-1, -2, -3, IgA, mucocidin, and α1-antitrypsin were more abundant in the late dominant SPC than the early dominant SPC, while histatin showed relatively even distribution in all SPCs. Therefore, it was presumed that the late dominant SPC containing abundant antimicrobial and protective proteins could be applied as a biomarker to measure the defensive potential of whole saliva in oral diseases.

A nineteen years old male patient showed a cystic lesion in left maxillary canine to premolar area (#23-#25). This lesion was asymptomatic, and found during his routine radiological check in local clinic. In the radiological observation the cystic lesion showed round radiolucent image containing many calcified bodies which were usually small but irregular in shape, expanding tumorously and resulted in the displacement of canine and second premolar in the absence of first premolar. The lesion was surgically enucleated, and a cystic fibrous tissue containing abnormal teeth was removed and examined pathologically. With the histological observation of tumorous odontogenic epithelium including many ghost cells, which were closely associated with abortive teeth, the lesion was finally diagnosed as CCOT associated with complex odontoma. The ghost cells of CCOT was strongly positive for β-catenin, GADD45, and LC3, and slightly positive for MMP-9, while they were rarely positive for BCL2, Wnt1, HSP-70, and p38. Therefore, it was presumed that the ghost cells of CCOT might undergo dormant cell state through altered cytodifferentiation stimulated by severe growth arrest, DNA damage signaling, and abundant autophage formation.

Aneurysmal bone cyst (ABC) in maxilla is a rare and benign lesion but shows extensive bony destruction, occasionally accompanied with secondary osseous lesions, i.e., central giant cell granuloma, ossifying fibroma, fibrous dysplasia, etc. As the pathogenesis of ABC has not been clearly defined, ABC is diagnostically challenged due to its variable histological features. A 17-year-old boy showed a huge radiolucent lesion at right anterior maxilla, which was accidentally found in routine dental-radiological examination for orthodontic treatment. He had no medical history of systemic disease, and did not remember any traumatic experience on his right anterior maxilla. The radiolucent lesion involved periapical area from right central incisor to right first premolar, and was clinically diagnosed as odontogenic keratocyst. During surgical operation a cyst-like sac was enucleated with severe hemorrhage. In the histological observation the thick fibrous sac showed no lining epithelium, and its luminal side disclosed multiple aneurysmal spaces which were shrunken and almost obliterated. The fibrous sac itself was hyperplastic with abundant vascular channels, and produced fibromatous thickening associated with ossifying trabecular bones. This fibro-osseous tissue was hamartomatous, which was not directly connected and organized with marrow bone of maxilla. Finally, the present case was diagnosed as secondary type ABC differentially from traumatic bone cyst (TBC), odontogenic cyst, and central reparative granuloma. And it was presumed that the hamartomatous proliferation of fibro-osseous tissue in the cystic sac of ABC could produce the swelling pressure effect in the bone marrow similar to the overgrowth of central giant cell granuloma, ossifying fibroma, fibrous dysplasia, etc., in the secondary type ABC.

Calcipex II has been widely used for root canal irrigation in endodontic treatment. It is a product of calcium oxide-based water soluble paste containing fine granular resin materials1). It was known that these granular materials were hardly dissolved in tissue and subsequently elicited foreign body granuloma by recruiting macrophages. Sometimes serious complications involving regional osteomyelitis and maxillary sinusitis were followed in long time after the endodontic treatment using Calcipex II materials2,3). And then the removed surgical specimen should be carefully examined to detect whether there exists Calcipex II material-related foreign body reaction or not. As the fine granular materials are too small in size, about 1 μm in diameter, and slightly translucent, it is difficult to find out the fine granular materials scattered throughout the granulomatous lesion even in the high magnification view. Here, we firstly found that the fine granular materials of Calcipex II showed bright birefringence under the polarizing microscope, and that the Calcipex II granules dispersed in chronic granulomatous lesion could be easily detected by their bright birefringence. The present study demonstrated a case of peri-implantitis involved with Calcipex II granule-related periapical granuloma, exhibiting numerous bright birefringence spots in the polarizing microscope observation.

An 81-years-old woman presented multiple mucosa ulcers with a chief complaint of pain during wearing the lower denture. She had been wearing upper and lower complete dentures for five months, and received multiple drugs for the treatment of angina pectoris, constipation, neurosis, hypertension and arthritis (calcium channel blockers, furosemide, captopril, nonsteroidal anti-inflammatory agents and penicillamine, respectively), but no history of immune-diseases and viral infection symptom. The present lesion was primarily diagnosed as traumatic ulcer, candidiasis and lichen planus in the clinical observation, thereby conservatively treated with denture relining, antifungal agent, and steroidal agent. However, the ulcer lesion was not healed for two months and rather increased in size. With the diagnosis of viral infection the immunohistochemical (IHC) staining of IL28 and E6, and polymerase chain reaction (PCR) using herpes simplex virus (HSV)-1 primer sets was done but entirely showed negative reaction. Therefore, with the patient’s medical history and IHC findings exhibiting strong positive reaction of CD3 and CD28, but rare/weak reaction of NFkB, CD20, IgK and p38, the ulcer lesion was finally diagnosed as drug-induced pemphigoid ulceration which was not an inflammatory granulomatous lesion but related to the retrogressive acantholytic degeneration of epithelial cells caused by multiple drug abuse.

In order to know the characteristic roles of salivary protein complex (SPC) the gel-filtration chromatography was performed using the unstimulated and the stimulated whole saliva separately. The first and second dominant SPC peaks were fractionated and analyzed by immunoprecipitation HPLC (IP-HPLC) using antibodies against the essential salivary proteins including α-amylase, mucin-1, proline rich proteins (PRPs), histatin, cystatin, LL-37, lysozyme, lactoferrin, -defensin-1, -2, -3, IgA, transglutaminase 4 (TGase 4), mucocidin, α1-antitrypsin, cathepsin G. In the gel-filtration chromatography the stimulated whole saliva showed much reduced amount of SPCs than the unstimulated whole saliva, but the proportional patterns of both whole saliva were almost similar each other. Through IP-HPLC analysis both of the first and second dominant SPCs were variably positive for the essential salivary proteins, however, α-amylase, mucin-1, PRPs, lysozyme, and cathepsin G were predominant in the first dominant SPC, while cystatin, lactoferrin, β-defensin-1, -2,-3, IgA, mucocidin, TGase 4, and α1-antitrypsin were predominant in the second dominant SPC. And more, the α1-antitrypsin and cathepsin G which were mostly derived from gingival crevicular fluid were also consistently found in the SPCs. These data may suggest that the first dominant SPC, rich in α-amylase, mucin-1, PRPs, lysozyme, and cathepsin G, may play a role in food digestion, protein degradation, and mucosa lubrication, while the second dominant SPC, rich in cystatin, lactoferrin, β-defensin-1, -2, -3, mucocidin, IgA, TGase 4, and α1-antitrypsin, may play a role in the mucosa protection and antimicrobial defense.

Caliber persistent artery (CPA) is a vascular anomaly presenting as a bluish and pulsatile artery in the subepithelial tissue. Although the incidence of CPA was debated, many CPAs occurred in the perioral and facial tissues at which the embryonal strapedial artery networks were distributed. The present study demonstrated a case of CPA occurred in the retromolar buccal mucosa in a 37 years old male. The lesion showed many pinkish granular spots, but was asymptomatic except biting irritation during mastication. It had slowly increased in size up to 20 × 25 mm for 3 years, and recently became hemorrhagic due to the biting injury between left upper and lower second molars. With the fear of oral cancer an incisional biopsy was performed, and followed by histological and immunohistochemical study. Histologically the lesion showed many tortuous artery localized at the submucosa area, and the arterial wall was thick and its lumen was narrowed and shrunken. In the immunochemistry α-SMA was positive for thick smooth muscle layer of artery and arterioles, TGase 2 was weakly positive for the luminal surface of arterial intima, and bFGF was consistently positive for the perivascular fibrous tissue. But PCNA, VEGF, CD31, CMG2, TGF-β1, HSP-70, and 14-3-3 were almost negative for the vascular tissue. Therefore, it was presumed that the lesion was not actively proliferative nor degenerative but still retained its cellular stability and slow growing potential. It was finally diagnosed as CPA differentially from arterio-venous malformation, hemangioma, lymphangioma, and squamous cell carcinoma. The retromolar buccal mucosa CPA is first reported in this study and may present usual clinical findings depending on its size and location. This asymptomatic lesion could be severely hemorrhagic by minor biting injury, therefore, precise differential diagnosis should be made through biopsy, and careful therapy be followed.

A 57 years old female received xenogenic bone graft for the extraction socket augmentation of right maxillary molars and for the sinus floor elevation six months ago. The bone graft sites were healed uneventfully and showed marked radiopacity in the postoperative X-ray view. Before dental implant insertion the bone biopsy was made using trephine bur and examined pathologically. The graft bones showed minimum new bone deposition with dysplastic epithelium. The epithelium was proliferative on the surface of graft bones forming epithelial strands and nests, similar to the odontogenic epithelium. The immunohistochemical study was performed using different antisera of odontogenic markers, growth factors, oncogenes, etc. The epithelial cells were strongly positive for pan-keratins, EGF, pAKT, and HSP-70, consistently positive for PCNA, p53, EGFR, 14-3-3, and survivin, slightly positive for ameloblastin, but rarely positive for amelogenin. Particularly the matrix of graft bone was slightly positive for EGF. Taken together, it is presumed that the abnormal epithelium on the graft bones was derived from odontogenic epithelial elements, Malassez epithelial rests, distributed at the periodontal tissue of maxillary molars, and that they might undergo dysplastic proliferation affected by the release of growth factors and osteogenic proteins from the graft bones. It is also suggested that the graft bone substitutes inserted for the dental implant possibly have a potential to induce the proliferation of odontogenic epithelial rests leading to the pathogenesis of odontogenic cysts and tumors.

Salivary proteins include numerous functional proteins which play important roles not only for the food-intake but also for the protective and defensive mechanisms. In the present study the compositions of salivary proteins were analyzed by different methods, including electrophoresis and high performance liquid chromatography (HPLC). In hydrophobic protein HPLC analysis the parotid saliva gradually produced macromolecular complexes when agitated in refrigerator until 30 minutes. These salivary protein complexes were digested by neuraminidase, and then migrated more rapidly in native tris glycine gel than the control. Therefore, it was assumed that the glycosylated proteins of parotid saliva became gradually aggregated to form salivary protein complexes similar to those of whole saliva. The salivary protein complexes were easily degenerated in different experimental buffers, i.e., SDS buffer, tris glycine buffer, methanol, etc., and resulted non-specific patterns in electrophoresis and HPLC. Therefore, it was presumed that the salivary protein complexes was made by the hydrophobic interaction as well as electrostatic attraction between salivary proteins. These data indicated that to know the real pattern of salivary protein complexes in vivo the whole saliva should be analyzed by HPLC using non-adhering column with isoelectric buffer. Consequently, the whole saliva was analyzed by HPLC using reverse phase SuperoseTM column with 20 mM potassium phosphate buffer, and two prominent peaks of salivary protein complexes were consistently found in every people. These salivary protein complex peaks were relatively stable up to 6 hours after saliva collection when the whole saliva was kept in refrigerator during experiment. Therefore, it is suggested that the salivary protein complex patterns are characteristic macromolecular structures of whole saliva, which are also applicable as a diagnostic point in different saliva-associated diseases

An 18 years old female patient suffered from cerebrovascular occlusive disease, moyamoya disease, showed a huge cyst in her left mandibular body in the radiological observation. The lesion was asymptomatic and found during routine dental check. She had no experience of traumatic injury on her jaw. The cystic lesion was ovoid with irregular scalloping margin and multilobular image, and occupied the whole marrow space of mandibular body with slight expansion of buccal cortical bone. During operation the lesion showed an empty space covered with grayish white fibrous tissue. The luminal fibrous tissue and underlying bony tissue were curettaged and examined pathologically. In the histological observation the lesion was a pseudocyst lined by thick fibrous tissue. Some large vessels underwent atherosclerotic change, exhibiting thickened vessel walls which were partly distorted with hemorrhage and thrombi, and some small capillaries were extremely dilated with hemorrhage and subsequently resulted in perivascular ischemic change with chronic vasculitis. This mandibular cystic lesion was finally diagnosed as simple bone cyst (SBC) associated with moyamoya disease differentially from aneurysmal bone cyst (ABC), traumatic bone cyst (TBC), periapical odontogenic keratocyst, and central giant cell granuloma. Therefore, it was presumed that the thromboembolic and atherosclerotic vessels of moyamoya disease might increase the hemodynamic pressure of mandibular bone marrow tissue and subsequently was able to induce SBC.

Oral galvanism is known to induce chronic irritation on oral mucosa, but the related pathology rarely occurs. A 65 years old male complained of linear horizontal ulcerations on his bilateral buccal mucosa for one month. The oral ulcerations were parallel and approximated to his occlusal plane. He had multiple metallic crowns using gold and nickel cobalt alloy at bilateral upper and lower molar teeth, and also had accustomed to heavy smoking for more than twenty years. Biopsy examination was performed with immunohistochemical staining using antisera of PCNA, D2-40, and PARP. The epithelial ulcer had clear margin and was replaced by granulation tissue containing many dilated lymphatic vessels, which were positive for D2-40, but showed no feature of pseudo-necrotic membrane. Nearby epithelium showed the typical features of leukoedema, characterized by edematous keratinocytes with clear cytoplasms and pyknotic nuclei, low rete ridges, and rough superficial epithelial layer, where PCNA was rarely positive. Some superficial edematous keratinocytes showed perinuclear cytoplasmic vacuolization, and their peripheral nuclear chromatins were positive for PARP. Taken together, the present mucosa ulceration was different from aphthous stomatitis, herpes stomititis, oral lichen planus, etc., thus it was postulated that the galvanic current generated from between the upper and lower dissimilar metal crowns could affect the precedent leukoedema caused by heavy smoking habit and produce the linear horizontal buccal ulceration. Therefore, the present case was diagnosed as galvanic mucositis associated with leukoedema, and it was also hypothesized that the mild and persistent galvanic current was able to deplete the cytoplasmic fluid of leukoedema keratinocytes via the osmotic pressure difference elicited by increased ionic concentration of galvanism, and followed by severe keratinocyte apoptosis and oral mucosa ulceration.

A 67 years old female showed diffuse erosive ulceration at left buccal mucosa. She had received tegretol to treat the patient’s pain and anxiety of trigeminal neuralgia for 18 months. Otherwise her medical history was nonspecific. Under the clinical diagnosis of lichen planus she received anti-inflammatory therapies using antibiotics and steroid ointment, which were not effective. Consequently her oral ulceration was gradually expanded and aggravated. In the biopsy examination mucosa epithelium was irregularly keratinized and focally detached from underlying connective tissue by thin cleft spaces, accompanied with inflammatory cell infiltration into the subepithelial area. The epithelium was generally acanthomatous with short rete ridges. Many spots of acantholysis were found in the basal and suprabasal layers of epithelium, into which melanocytes were migrated. Particularly, many keratinocytes not only in the spinous layer but also in the suprabasal layer contained atypical keratohyalin granules in their cytoplasms. In the immunohistochemistry the epithelium was rarely positive for PCNA and IgK, but strongly positive for HSP-70, and many keratinocytes showed strong positive reaction of lysozyme in their cytoplasms. Taken together, with the characteristic cytotoxic changes of keratinocytes, which are usually found in the oral epithelium damaged by certain drug abuse, the present case of pemphigus-like oral lesion was diagnosed as drug-induced pemphigus caused by long time intake of tegretol, carbamazepine derivative. The acute oral drug-induced pemphigus should be differentially diagnosed from oral lichen planus, recurrent aphthous ulceration, oral leukoplakia, candidiasis, autoimmune pemphigus, etc., in order to treat properly in the absence of biohazards of systemic therapeutic drugs