Lymph node hyalinization in elderly Japanese.

Lymph node hyalinization has been comprehensively investigated using specimens obtained from elderly Japanese and white Americans. Onion-peel lesions and associated meshwork areas were often found in the medullary sinus of the thoracic node (mediastinal-type hyalinization), while eosinophilic, glassy and spotty lesions were consistently seen in B lymphocyte areas of the pelvic node (pelvic-type hyalinization). The mediastinal-type hyalinization was comprised of thin collagen fibrils (ca 50 nm in diameter), whereas the pelvic-type hyalinization had thick fibrils (ca 150 nm in diameter). This difference seemed to be consistent with a difference in composite collagen fibrils of vascular walls between the thoracic and pelvic regions. The pelvic-type hyalinization was often or sometimes seen in other nodes, such as cervical, axillary, abdominal and inguinal nodes, especially in white Americans. The mediastinal-type hyalinization, usually in combination with a sinus filled with anthracotic macrophages, tended to be observed in Japanese more frequently than in white Americans. Anthracosis seemed to be connected to the pathogenesis of the hyalinization. On the other hand, because the lesion was weakly positive for Factor VIII immunohistochemistry and because lesions were located along thin vessels, the pelvic-type hyalinization seemed to originate from vascular degeneration in the nodal cortex. Due to the high incidence and large proportion in total volume of the node, the hyalinization seems to be one of the major events that diminish the nodal filtration function and ruin the node with aging.

Interferon-gamma downregulates Hsp27 expression and suppresses the negative regulation of cell death in oral squamous cell carcinoma lines.

Interferon-gamma (IFN-gamma) induced cell death in five oral squamous cell carcinoma (SCC) lines. Cell death was specific to IFN-gamma treatment and did not occur with either IFN-alpha or TNF-alpha. IFN-gamma did not induce typical apoptotic phenotype in cells, such as morphological changes and DNA ladder formation. Caspase-3 was partially activated by IFN-gamma. Protein levels of molecular chaperones were examined in cells treated with IFN-gamma. Among these, levels of heat shock protein 27 (Hsp27) were specifically reduced upon IFN-gamma treatment of oral SCC cells. Recombinant clones overexpressing Hsp27 were more resistant to IFN-gamma-induced cell death than parent cells. Conversely, cells expressing a dominant-negative mutant of Hsp27, in which three serine residues (15, 78 and 82) were replaced by glycine, were hypersensitive to the effects of IFN-gamma and exhibited a typical apoptotic phenotype. Pretreatment of cells with IFN-gamma enhanced apoptotic cell death induced by cisplatin. Our data suggest that IFN-gamma suppresses Hsp27 expression in oral SCC cells and blocks the inhibitory effects of this molecular chaperone on apoptotic cell death. Moreover, IFN-gamma initiates the transition of oral SCC cells to the proapoptotic and/or aborted apoptotic state. Hsp27 plays a crucial role in the inhibition of apoptosis of oral SCC cells. Our findings highlight the importance of employing IFN-gamma in combination with certain anticancer drugs as treatments for oral cancer. We suggest that Hsp27 plays a significant role in the IFN-gamma-induced sensitization of oral SCC cells to anticancer drugs.

Changes in occlusal state of patients with mandibular prognathism after orthognathic surgery: a pilot study.

This study examined the occlusal state of patients with mandibular prognathism and compared it with that of adults with normal occlusion (controls). It also examined changes in occlusal state after orthognathic operations in these patients. The values of occlusal contact area and bite force in patients before operation were significantly lower than in controls, and occlusal pressure in patients was higher than in controls. The occlusal contact area and bite force of the patients 1 month after the operation had decreased to below preoperative values. These values 12 months after the operation had increased by 2.0 and 1.8 times in women and 1.4 and 1.4 times in men, respectively, compared with preoperative values. However, absolute values remained extremely low compared with those of controls. In contrast to the above, occlusal pressure reached its maximum value 1 month after the operation and at 12 months it was close to the value for controls.

pRb2/p130 protein expression is correlated with clinicopathologic findings in patients with oral squamous cell carcinoma.

BACKGROUND: pRb2/p130 is one of the retinoblastoma (Rb) gene family and a suppressor oncogene. Immunohistochemically, the expression of pRb2/p130 was reported to be correlated inversely with the degree of malignancy in lung carcinoma and endometrial carcinoma. In the current study, the correlation between expression of pRb2/p130 and clinicopathologic factors in oral squamous cell carcinoma was investigated. METHODS: One hundred twenty-two specimens from patients with oral squamous cell carcinoma were investigated by staining with a polyclonal antibody against pRb2/p130. The correlation between the expression of pRb2/p130 and various clinicopathologic factors was studied. RESULTS: Positive staining for pRb2/p130 was observed in 61 of 122 cases (50.0%). pRb2/p130 expression was found to be correlated significantly with clinical stage (P = 0.050), cervical lymph node metastasis (P = 0.035), and tumor differentiation (P = 0.050). In the entire group a significantly reduced 5-year cumulative survival rate was observed in patients with pRb2/p130-negative tumors compared with patients whose tumors positively expressed pRb2/p130 (P = 0.0004). When tested with Cox proportional hazards regression analysis, the most significant independent prognostic factor for the entire group of 122 patients was found to be pRb2/p130 expression. CONCLUSIONS: Expression of pRb2/p130 may be a good prognostic indicator in patients with oral squamous cell carcinoma and also may be utilized for the subclassification of tumors with the Grade 3 mode of carcinoma invasion.

Expression of Rb, pRb2/p130, p53, and p16 proteins in malignant melanoma of oral mucosa.

We previously reported that pRb2/p130 gene, one of the Rb family members, was immunohistochemically abundantly expressed in well-differentiated oral squamous cell carcinomas, whereas in undifferentiated ones the expression was low. Oral malignant melanoma is extremely rare, however the prognosis is poor because it tends to locally invade tissue or metastasize and its biological behavior appears to be different from cutaneous malignant melanoma. The present study dealt with the expression of pRb2/p130, Rb, p53, and p16 in 13 cases of malignant melanoma of oral mucosa as revealed by immunohistochemical staining. The stage classification of the 13 patients was as follows; stage II: eight patients, stage III: three patients, and stage IV: two patients. pRb2/p130 was expressed in only two stage II-cases, neither of which have shown any evidence of recurrence or metastasis for over 14 years. Positive staining for Rb was found in three cases consisting of one stage II-case, one stage III-case, and one stage IV-case. p53 was expressed in two cases, one a stage II and the other a stage IV. Positive staining for p16 was found in seven cases consisting of four stage II-cases, two stage III-cases, and one stage IV-case. pRb2/p130 may be inversely correlated with the malignancy of oral malignant melanoma, but further study is needed.

Retinoid X receptor alpha and retinoic acid receptor gamma mediate expression of genes encoding tight-junction proteins and barrier function in F9 cells during visceral endodermal differentiation.

Retinoids are critical for differentiation of columnar epithelial cells and for preventing metaplasia of these cells into stratified squamous epithelial cells, in which tight junctions (TJs) are essentially absent. This implies that retinoids might play important roles in regulating the structures and functions of TJs of columnar epithelium. F9 murine embryonal carcinoma cells differentiate into epithelial cells resembling visceral endoderm bearing TJs, when grown in suspension as aggregates in the presence of retinoic acid (RA). We show that RA induces the TJ structure and expression of several TJ-associated molecules, such as ZO-1, occludin, claudin-6, and claudin-7, as well as a barrier function in the genetically engineered cell line F9:rtTA:Cre-ER(T) L32T2, which allows sophisticated genetic manipulations simply by addition of ligands (H. Chiba et al., 2000, Exp. Cell Res. 260, 334-339). Interestingly, our data indicate that a barrier for small substances is generated after that for large ones during de novo formation of TJs. We also compared the RA-induced expression of TJ components and barrier function in RXRalpha(-/-)-RARgamma(-/-) F9 cells with those in wild-type cells and show that the retinoid signals for transduction of these events are mediated by specific RXR-RAR pairs.

Incidence of cleft lip or palate in 303738 Japanese babies born between 1994 and 1995.

To investigate the incidence of cleft lip or palate or both (CLP) in Japan, 303738 babies born in 1532 institutions between 1994 and 1995 were examined and 437 (0.14%) were found to have abnormalities. Of these babies, 32.1% had cleft lip, 43.3% had cleft lip and palate, and 24.8% had cleft palate (Table 2). These results show that the incidence of cleft lip and palate has declined compared with the period from 1981 to 1982.

Mumps virus can suppress the effective augmentation of HPC-induced apoptosis by IFN-gamma through disruption of IFN signaling in U937 cells.

Cells of the human promonocytic cell line U937 were found to be sensitive to hexadecylphosphocholine (HPC), which is a potential anticancer drug. Induction of apoptosis was found in U937 cells after treatment with HPC for 24 to 48 hr. The apoptosis in U937 cells exposed to HPC was increased significantly in the presence of interferon-gamma (IFN-gamma). The augmentation of HPC-induced apoptosis by IFN-gamma is repressed in cells (U937-MP) persistently infected with mumps virus. A persistently infected cell line, U937-MP, showed poor induction of signal transducers and activators of transcription-1alpha (STAT-alpha), STAT-2, p48 and IFN-regulatory factor-1 (IRF-1), which are closely correlated with interferon-alpha (IFN-alpha) and IFN-gamma signaling pathways. Expression of MHC class-I or class-II was augmented by IFN-alpha or IFN-gamma in U937 cells, but not in persistently infected cells. Therefore, it is suggested that the IFN-gamma signaling pathway plays an important role in the augmentation of HPC-induced apoptosis. Mumps virus can suppress the IFN-gamma signaling pathway and subsequent development of apoptosis.

Comparative study of oral squamous cell carcinoma in Okinawa, Southern Japan and Sapporo in Hokkaido, Northern Japan; with special reference to human papillomavirus and Epstein-Barr virus infection.

In Okinawa, a subtropical island in Southern Japan, the incidence of oral squamous cell carcinoma is 1.5 times higher than that in mainland Japan. Sixty cases of oral squamous cell carcinoma from 1993 to 1996 in Okinawa and 42 cases over the same period in Sapporo were examined histologically. Human papillomavirus (HPV) and Epstein-Barr virus (EBV) were detected by polymerase chain reaction (PCR) amplification with primers specific for HPV and EBV. In situ hybridisations of the viruses were also carried out. In the case of Epstein-Barr virus, in situ PCR was also performed. Thirty-five (58.3%) Okinawan tumours were well-differentiated in type, but in Sapporo, 18 (42%) were of such type. In Okinawa, tumours of the mouth floor (10 cases, 16.7%) and oropharynx (12 cases, 20%) were frequently observed, whereas in Sapporo only five cases (12%) of each were found. HPV was demonstrated in 78% of Okinawan cases and 26.2% of Sapporon cases by PCR or non-isotopic in situ hybridisation (NISH). There were 76.6% (46 cases) of Okinawan and 38.1% (16 cases) of Sapporo cases positive for EBV by PCR. In only 12 Okinawan cases and 4 Sapporon cases, were positive signals demonstrated by in situ PCR on the cancer cells themselves. EBV was demonstrated in the large number of infiltrating lymphocytes, most of which were CD3+, and a few were CD19+. In Okinawa, HPV might be an important causative factor of oral squamous cell carcinoma and EBV a less important factor, whereas in Sapporo HPV and EBV might play only a small part in the aetiology of the tumour.

Clinical features and management of oral and maxillofacial tumors in children.

OBJECTIVE: To clarify the characteristics of oral and maxillofacial tumors in children, we carried out a clinical study of such tumors and reviewed the relevant procedures for treatment. Study design. Medical records of 105 patients less than 15 years of age who were treated for oral and maxillofacial tumors during the 20 years between 1976 and 1997 were retrieved and analyzed. RESULTS: Of the 105 cases, 102 (97.1%) involved tumors that were benign; only 3 patients (2.9%) had tumors that were malignant. With regard to benign soft tissue tumor, the most common type was hemangioma (25/69; 36.2%), the second most common type was papilloma (19/69; 27.5%), and the most common site was the tongue. With regard to bone tumor, the most common type was odontoma (14/33; 42.4%), the second most common type was ameloblastoma (11/33; 33.3%), and the most common site was the mandible. Most of the odontogenic tumors (25/28; 89.3%) developed in patients more than 6 years of age; these tumors may develop after dental crown formation. In each case of benign soft tissue tumor, resection was performed; 4 of these tumors (2 hemangiomas, 1 lymphangioma, and 1 papilloma) recurred, but after re-resection recurrence has not been found for more than 4 years. In the cases of benign jawbone tumor, resection and enucleation were the procedures principally performed; 3 ameloblastomas recurred after enucleation, but after relatively wide resection for these recurrent tumors there has been no recurrence for more than 17 years. With regard to the ameloblastomas, enucleation with preservation of the periosteum was effective and bone regeneration occurred rapidly. CONCLUSIONS: Most oral and maxillofacial tumors in children are benign. In any case of such benign tumor, including ameloblastoma, minimal surgical treatment should be the procedure of first choice.

Augmentation of verotoxin-induced cytotoxicity/apoptosis by interferon is repressed in cells persistently infected with mumps virus.

Verotoxin type 2 (VT2) produced by enterohemorrhagic Escherichia coli (EHEC) has been shown to have high cytotoxic potency toward several human B lymphoid cell lines with and without Epstein-Barr virus (EBV). Cell death, apoptosis induced by VT2, is closely correlated with the expression of receptor molecule Gb3/CD77, recognized by the toxin, but not with the infection or presence of EBV. Pretreatment of cells with interferon-alpha (IFN-alpha) for 24 h resulted in augmentation of apoptosis by VT2. Pretreatment within 8 h, however, was not effective. It has been reported that IFN-alpha-induced apoptosis is correlated with the induction of the 2',5'-OAS/RNase L system or dsRNA-activated protein kinase (PKR) or both. We have established persistent infection in both Akata and P3HR-1 cells with mumps virus. The persistently infected cell lines, P3HR-MP2 and Akata-MP2, showed poor induction of 2',5'-OAS and PKR in response to IFN-alpha. Augmentation of VT2-induced apoptosis by IFN-alpha was not found in the cell lines P3HR-MP2 and Akata-MP2. Therefore, these findings were interpreted to indicate that augmentation of VT2-induced apoptosis by IFN-alpha may be mediated by PKR and the 2',5'-OAS/RNaseL system. It is also suggested that mumps virus can suppress apoptosis and establish persistent infection.

Immunohistochemical investigation of new suppressor oncogene p130 in oral squamous cell carcinoma.

New onco-suppressor gene p130 is one of the Rb family forms and is reported to undergo allelic loss in hepatocellular, prostate, and breast carcinomas; however, no report of p130 has been made in oral tumors. The present study dealt with the expression of p130 and Rb proteins by immunohistochemical staining in oral squamous cell carcinomas (n = 110) and oral mucosa. The site of the carcinomas included tongue (n = 48), gingiva (n = 32), oral floor of the mouth (n = 15), oropharynx (n = 6), buccal mucosa (n = 5), and others (n = 4). Histologically there were 65 well-differentiated carcinomas, 33 moderately differentiated carcinomas, and 12 poorly differentiated ones. Positive staining for p130 and Rb was localized to suprabasal cell layers of the normal oral epithelium. In the oral squamous cell carcinoma, positive staining for p130 and Rb was observed in well-differentiated carcinomas (p130, 66.2%; Rb, 78.5%), more than in poorly differentiated ones (p130, 16.7%; Rb, 50.0%). In immunoelectron microscopic features, p130 protein was localized in the nucleus and mitochondria. The expression of p130 was related to the degree of tumor differentiation as that of Rb. It is suggested that p130 gene may be associated with the development of a wide variety of human malignancies rather than the progression.

Prognostic factors and relative risk for survival in N1-3 oral squamous cell carcinoma: a multivariate analysis using Cox's hazard model.

The records of 136 patients with N1-3 oral squamous cell carcinoma treated by surgery were investigated retrospectively, with the aim of finding out which factors were predictive of survival on multivariate analysis. Four independent factors significantly influenced survival in the following order: pN stage; T stage; histological grade; and N stage. The most significant was pN stage, the five-year survival for patients with pN0 being 91% and for patients with pN1-3 41%. A further study was carried out on the 80 patients with pN1-3 to find out their prognostic factors for survival and the independent factors identified by multivariate analysis were T stage and presence or absence of extracapsular spread to metastatic lymph nodes.

Oral malignant melanoma: long-term follow up in three patients.

Three patients with oral malignant melanoma, who survived for at least ten years after initial examination, are presented. The depth of tumor invasion was 5 mm or less in all patients. The first patient received surgery and postoperative immunotherapy and she had no recurrence more than 18 years after treatment. The second patient underwent surgery but had local recurrence after 11 years. He underwent resection and postoperative chemotherapy and was still alive more than 14 years after initial examination. The third patient received radiotherapy and had metastasis nine months after treatment. Resection and radiotherapy were performed and the patient was alive more than 14 years after first presentation.

Multivariate analysis of occult lymph node metastasis as a prognostic indicator for patients with squamous cell carcinoma of the oral cavity.

BACKGROUND: The biologic aggressiveness of squamous cell carcinoma of the oral cavity is reflected in its ability to metastasize to regional cervical lymph nodes. Patients with clinically negative cervical lymph nodes are believed to have a good prognosis; however, the prognosis of patients with lymph node metastasis occurring after excision or radiotherapy of the primary tumor is poor. METHODS: Univariate and multivariate analyses for occult lymph node metastasis (ONM) in 172 patients with clinically negative cervical lymph nodes were performed by the authors to elucidate the clinical and histologic tumor risk factors to enhance their ability to predict ONM. A multivariate Cox proportional hazards model and Hayashi's quantification theory type II were used to analyze prognostic factors and to determine the probability of ONM. RESULTS: Using Cox's proportional regression model, the factors linked to cancer specific survival were selected: tumor differentiation (P = 0.0330), mode of carcinoma invasion (P = 0.0175), and ONM (P = 0.0433). Pathologically identified metastatic lymph nodes were found in 21.5% of the cases studied (37 of 172 cases). The 5-year cancer specific survival was 94.0% for patients without lymph node metastasis, and 51.0% for patients with ONM (P < 0.0001, log rank test). The most significant predictors for ONM of each of the clinical and histologic factors, in descending order, were: mode of carcinoma invasion, intensity of lymphocytic infiltration, degree of differentiation, number of mitotic figures, and type of growth by means of Hayashi's quantification theory type II. The presence or absence of ONM in 147 of 172 patients (85.5%) was correctly predicted by the score at the point of intersection of the two curves, which was -0.03. Further investigation revealed that 28 of 32 new cases were differentiated accurately by means of this diagnostic system. CONCLUSIONS: The results of the current study suggest that this method of analysis can establish a reliable predictor of ONM, thereby facilitating correct choices for surgical procedures to enhance the survival rates of patients with clinically negative cervical lymph nodes.

Cytotoxicity of histocompatibility leukocyte antigen-DR8-restricted CD4 killer T cells against human autologous squamous cell carcinoma.

Although CD8+ killer T cells reacting against human autologous tumor cells have recently been studied in detail, little is known about the cytotoxic mechanism of CD4+ T cells against such tumor cells. In order to investigate this, we have established CD4+ cytotoxic T lymphocyte TcOSC-20 lines. TcOSC-20 showed selective cytotoxic activity against autologous OSC-20 cells, derived from a cancer of the tongue, in an HLA-DR-restricted fashion. HLA-DR8 (DRB1*08032) is the only DR molecule expressed on OSC-20 cells, and anti-DR8 monoclonal antibody could inhibit the cytotoxicity, suggesting that HLA-DRB1*08032 is the tumor rejection antigen-presenting molecule to TcOSC-20. The Fas ligand was expressed on TcOSC-20 lines, and its expression was induced upon mixed lymphocyte-tumor cell culture of autologous peripheral blood lymphocytes. Furthermore, the cytotoxicity of TcOSC-20 was inhibited by anti-Fas ligand antibody. These data imply that TcOSC-20 lines recognize the tumor antigenic peptide presented by HLA-DR8, and exert cytotoxicity against autologous tumor cells via a Fas-mediated cytotoxic pathway.

Supraperiosteal flap technique versus mucoperiosteal flap technique in cleft palate surgery.

Recent animal experiments have shown that palatal repair without denudation of bone leads to a superior dento-alveolar development. The aim of this clinical study was to evaluate the peri- and postoperative course and the dento-alveolar development of the deciduous dentition in Japanese ULCP, and CP patients up to 5 years after two different types of palatal repair. One of the methods, the Kohama (1991) supraperiosteal flap technique, is performed without denudation of the bony palate, while the other, the Wardill (1937) push-back technique, results in areas of denuded bone. It was concluded that the supraperiosteal technique can be performed successfully in approximately the same amount of time as the push-back technique. Re-epithelialization of the wound areas after supraperiosteal repair takes about 1 week, which is one third of the time associated with healing after the push-back technique. Arch depth of the deciduous dentition after the supraperiosteal technique is superior compared to the push-back technique. The question of whether or not the supraperiosteal technique produces more favorable dento-alveolar development than the mucoperiosteal technique in the permanent dentition in humans has to be elucidated in future research.

Maxillofacial fractures sustained during sports.

PURPOSE: This study investigated the number and type of maxillofacial fractures caused by various athletic activities. PATIENTS AND METHODS: The ninety-eight patients were treated between 1977 and 1993, and the type of sport involved, patient age and sex, cause of accident, site of injury, and mode of treatment were evaluated. RESULTS: Sports-related maxillofacial fractures accounted for 10.4% of all patients with facial bone fractures. The number of different sports was 19, with the incidence of the fractures being most common in rugby and skiing, followed by baseball and soccer. The ratio of males to females was 5.5:1, and most of the patients were between 10 and 29 years of age. Total restriction from sports activity was between 8 and 12 weeks after initial treatment. CONCLUSION: The treatment for sports-related maxillofacial fractures is not different from that for fractures from other causes. However, it is important to establish some standard for deciding the time when it is possible to participate in sports after a fracture.