An effect of aging factor was investingated on correlations between electrolyte concentrations of the human resting saliva and radial bone mineral density (BMD). The following results were obtained. Subjects, who consented to these measurements, were 164 nontreated plus control patients and 1480 steoporotic patients treated with the drug for improving bone metabolism. The BMD was measured by dual-energy X-ray absorptiometry at the distal of the radius. Salivary electrolyte and the related indices were measured by means of thin layer sampling.

1. A significant negative correlation between the radial BMD and age was confirmed in both female groups of the medicated-and nonmedicated-ones even after removal of the salivary electrolyte factors.

2. Nonsignificant correlations were found between the age vs. salivary pH₁ and △pH₁ in both female groups of the medicated-and nonmedicated-groups.

3. Although nonsignificant in the medicated female group, positive correlations between the radial BMD vs. the salivary pH₁ and △pH₁ were significant in the nonmedicated female group. Significance for these coefficients remained even after removal of the aging factor.

4. In the nonmedicated female group, there was a significant negative correlation between the radial BMD and the salivary [K⁺] even after removal of the aging factor.

5. In male groups, a significant negative correlation was found between the radial BMD and the salivary [K⁺] only in the medicated one, and this remained even after removal of aging component.

The localization of Na/K-ATPase in the mouse molar enamel organs and the subunit constitution of the enzyme were investigated by means of enzyme histochemistry and immunohistochemistry, using ahtibodies for the α and β subunits of the anti-rat Na/K-ATPase. Intense enzyme reaction of ouabain-sensitive p-nitrophenylphosphatase was specifically localized on the stratum intermedium. Among five different anti-subunits of Na/K-ATPase, only α₂ and β₁ subunits were immunohistochemically localized on the stratum intermedium. When the chronological change in the localization of Na/K-ATPase during tooth development was immunohistochemically assessed using anti-β₁ subunit, the area of positive reaction in stratum intermedium moved from the cuspal region to the cervical region as the enamel matured. After the completion of the enamel matrix maturation, the immunoreaction for the anti-β₁ subunit disappeared from the enamel organ. These results indicate that Na/K-ATPase in the stratum intermedium is composed of α₂ and　β₁ subunits, and that the enzyme is significantly involved in the enamei maturation.

There are some reports that activity in the masticatory system affected the growing craniofacial skeleton. We speculated that biting efficiency affects not only cephalometric data but also mandibular bone mineral content (BMC). In this study, we evaluated the relationship between mandibular BMC and maximum bite force or occlusal contact area as a parameter of biting efficiency. The subjects consisted of 46 adults, with a mean age of 23 years and 7 months. Mandibular BMC was measured by photodensitometry using dental X-ray films. The sublect contrast of mandible was used as a parameter of BMC. Bite force and occlusal contact area were measured using pressure sensitive sheets. Anegative correlation was observed between subject contrast and bite force and between subject contrast and occlusal contact area, with a correlation coefficients of -0.378(p<0.01) and -0.401(p0.01) respectively. Thus, BMC increased with maximum bite force and occlusal contact area. It was indicated that masticatory activity appears to affect not only cephalometric data, as previously reported, but also bone strength.

Effects of neuropeptides, cholecystokinin-octapeptide (CCK-8:5,50,500 μg/kg, s. c.) and its analogue ceruletide (Cer: 0.4,4,40 μg/㎏, s. c.), on the salivary responses induced by autonomimetic drugs in mice were analyzed in a pharmacological manner.

It was found that CCK-8 further stimulated an increased response to the pilocarpine (0,8 ㎎/kg, s.c.) -induced saivation in the cholinergic nervous system. Cer further stimulated response to the dobutamine (10 ㎎/kg, s. c.) -induced salivation, but not to salbutamol (40 ㎎/㎏, s. c.), in the adrenergic nervous system. On the other hand, CCK-8 and Cer showed no effects on the phenylephrine (5 ㎎/kg, s. c.) -and clonidine (5 ㎎/kg, s. c.) -induced salivation in the adrenergic nervous system. Thus, it suggests that CCK-8 has a different effect compared to Cer on the responses to the salivary secretion induced by autonomimetic drugs in mice.

Since angiogenesis is critical to the proliferation of solid tumors, the inhibition of this process may provide a new approach for cancer therapy. TNP-470 (O- (chloroacetyl-carbamoyl) fumagillol), a novel semisynthetic analogue of fumagillin isolated from. Aspergillus fumigatus, has potential as an antiangiogenic agent, which selectively prevents endothelial cell proliferation through inhibition of DNA synthesised. In the present study, the effects of TNP-4700n tumor-induced angiogenesis, tumor proliferation and experimental pulmonary metastasis were examined using a weakly immunogenic squamous cell carcinoma of the WHT/Ht mouse. TNP-470 inhibited the angiogenesis induced by the murine tumor dose-dependently, and the number of blood vessels orientated towards the intradermal tumor was maximally reduced by 45%compared to the control. In the tumor proliferation assay, TNP-470 significantly inhibited proliferation of the subcutaneously inoculated tumor, and reduced the tumor volume by 29% (80㎎/kg) and 46% (40㎎/kg) compared to the control. A similar inhibitory effect on tumor proliferation was observed in the experimental pulmonary metastasis assay, and the number of metastatic foci (diameter≧1.0) after intravenous inoculation of tumor cells was lowered by 13% (80㎎/㎏) and 42% (40㎎/kg) compared to the control with injection of TNP-470. These results suggest that TNP-470 has strong inhibitory activities against in vivo tumor-induced angiogenesis and proliferation of murine squamous cell carcinoma at the subcutaneous and rnetastatic sites.

It is known that lymphatic capillaries surrounding tumors play a functional important role in invasion and metastasis of the cancer. In this study, absorbing substances from tissues by lymphatic capillaries around the tumor was evaluated in VX2 tongue carcinoma using horseradish peroxidase (HRP) as a tracer. Subjects were rabbits which were 7 and 14 days after VX2 carcinoma transplantation into their tongues. HRP was infused under the mucosa of the tongue at the site of tumor transplantation, and specimens for transmission electron microscopy were prepared. HRP was absorbed by lymphatic capillary endothelial cells from surrounding tissues through intercellular endothelial spaces and pinocytotic vesicles. More vesicles were observed in 7 and l4 days after transplantation of VX2 carcinoma than in the control group. Also, the amount of HRP transferred from the abluminal side of endothelial cells through intercellular endothelial spaces and accumulated on the cell membrane of the luminal side increased after transplantation. The number of pinocytotic vesicles increased in endothelial cells in the transplanted groups as compared with the control group, and the length of adhesion apparatus in intercellular endothelial spaces was reduced. These results suggested that the transport of substances especially via intercellular endothelial spaces and vesicles increased with progression of tumor in lymphatic capillary surrounding tumor.

This study evaluated the relationship between the growth of transplanted rabbit VX-2tongue cancer and the changes of blood flow in a lymph node with primary metastasis. Rabbits were anesthetized by injecting 25mg/kg pentobarbital sodium into the auricular vein. The gross tumor size and the blood flow in the tongues and the deep cervical lymph node of 5 animals each of the non-transplanted group and transplanted groups were measured 7,14 and 21 days after transplantation of VX2 cancer, and their relationships were evaluated. Newly formed vessels around the transplanted tongue cancer were eδamined through scanning electron microscopy of vascular casts. The blood flow was measured using a laser flowmeter (ALF21, ADVANCE).

The volume of the transplanted tumor and the size of the deep cervical lymph node increased progressively. The blood flow of tongue did not increase significantly 7 days after transplantation, compared with the non-transplanted group, but it increased significantly after 14 days and decrease significantly after 21days. The blood flow of the deep cervical lymph node did not increase significantly 7 days after transplantation, compared with the normal group, but it increased significantly after 14 and 21 days. Scanning electron microscopy of vascular cast specimens showed defects of the vasculature and newly formed vascular networks at the sites of cancer cell proliferation. Positive correlations were observed between tumor volume and tongue blood flow; deep cervical lymph node (L. N.) blood flow and tumor volume; the shortest diameter/ the longest diameter ratio of the deep cervical lymph node (S/L) and tumor volume; S/L and tongue blood flow; S/L and L. N. blood flow; tongue blood flow and L. N. blood flow. These results suggest that daily measurement of the blood flow of transplanted VX2 tongue cancer allows estimation of the presence or absence of metastasis in the deep cervical lymph node.

The isolation medium for Abiotrophia, the isolation ratio of Abiotrophia, from saliva and dental plaque of healthy adults, production of dextran and protease, and antimicrobial susceptibility of Abiotrophia were studied. The most appropriate medium for isolation of Abiotrophia was a double-layer agar plate made with a base layer of Columbia agar and a top layer of Columbia agar containing heat-killed Micrococcus luteus to reach the optical density of 2.O units, pyridoxal and cysteine. Samples were taken from 93 healthy young human. Out of these human subjects, 72 strains of Abiotrophia (77.4%) were isolated from saliva and dental plaque each. No isolation was observed in three human subjects. This suggests that Abiotrophia consists of normal flora in oral cavity. When 423 isolated Abiotrophia strains were identified, 192 of the 212 strains from saliva were A. adiacens (90.6%), and 20 (9.4%) were A. defectiva. Of the 211 strains from dental plaque, 180 were A. adiacens (85.3%) and 31 (14.7%) were A. defectiva. All of the strains produced dextran, and 28.7% of of A. adiacens and 66.7% of A. defectiva produced protease. By the examination of the antimicrobial susceptibilities of 22 agents of the 92 strains of Abiotrophia. both A. adiacens and A, defectiva were high susceptible to antimicrobial agents tested: benzylpenicillin, ampicillin, amoxicillin, aspoxicillin, cephalexin, cefazolin, cefotaxime, cefpirome, flomoxef, erythromycin, leucomycin and clindamycin. To imipenem/cilastain, streptomycin, gentamicin, lincomycin and vancomycin, A. adiacens were lower than A. defectiva. Minimum bactericidal concentration/minimum inhibitory concentration ratio ≧ 2 occurred in benzylpenicillin. This indicates no penicillin tolerant strains were found.

The activity of cellular proliferation was immunohistochemically studied in 26 hyperkeratoses (HK), 25 epithelial dysplasias (ED), 49 squamous cell carcinomas (SCC) and 24 states after treatment of SCC (t-SCC) of the oral cavity.These lesions were compared with normal mucosa adjacent to them.

The tissues of these lesions were fixed in neutral phosphate buffered formalin and embedded in paraffin. Serial sections cut at 3μm were mounted on silane-coated glass slides. The monoclonal mouse antibody against human proliferating cell nuclear antigen (PCNA, PC10) and polyclonal rabbit antibody against human p53 (CM-1) were used as primary antibodies. The deparaffinized sections were immunostained by streptoavidin-biotin complex methods using Pathostain ABC kit.

The numbers of positive cells were counted by using a computer-accessed image analyzer (IBAS-2000), and then calculated appearance index and labeling index. In each lesion, the appearance indexes(%) of PCNA and p53 positive cells were 32.0 and 24.0 in normal mucosa, 72.9 and 26.9 in HK,80.0 and 40.0 in ED, and 83.7 and 53.1 in SCC, respectively. Labeling indexes of PCNA and p53 positive cells were 2.4±0.9 and l.7±0.7 in normal mucosa, 9.7±8.9 and 4.8±1.8 in HK, 20.4±9.2 and 11.6±2.0 in ED and 22.6±10.3, and 24.8±11.0 in SCC, respectively. The ratio of p53-positive cells to 100tumor cells was significantly higher in SCC than any other lesions (ρ<0.01). In t-SCC, the appearance and labeling indexes (%) of p53 positive cells were 50.0 and 20.3±13.8, respectively.

These results suggested that p53 tumor suppressor gene product was a beneficical marker for estimating malignant formation in lesions of the oral cavity.

Pathological examinations undertaken in 1995 at the Department of Oral Pathology, Iwate Medical University, were statistically reviewed.The number of biopsy meteriales examined amounted to 722 (including 83 requested from outside sources) from 577 cases. According to histological classifications, odontogenic benign lesions consisted of 5 ameloblastomas, 3 periapical cemental dysplasias, 20 dontomas, 1 squamous odontogenic tumor and l adenomatoid odontogenic tumor. Of nonodontogenic benign lesions, 33 were diagnosed as fibromas or fibroma like lesions, 10 as hemangiomas, 9 as papillomas, 8 as lichen planus, 7 as foreign body granulomas, 4 as pleomorphic adenomas, 3 as osteomas, 2 as lipomas, and 2were neurofibromas. The malignant tumors consisted of 53 squamous cell carcinomas, 3 adenocarcinomas, and 4 other types. The odontogenic cyst consisted of 49 radicular cysts, 19 primordial cysts and 8 dentigerous cysts. Of nonodontogenic cysts, 27 were diagnosed as postoperative maxillary cysts, 25 as salivary gland cysts, 3 as simple bone cysts, 3 as epidermoid cysts, 2 as dermoid cysts and 2 incisive canal cysts. Also found were 26 cases of hyperkeratosis (leukoplakia), 10 epithelial dysplasias, 17 chronic and localized hyperplastic gingivitis (epulis), 31 with Sjögren syndrome, and 60 having chronic inflammatory (glanulation) tissue.