Advanced glycation end-products increase IL-6 and ICAM-1 expression via RAGE, MAPK and NF-κB pathways in human gingival fibroblasts.

BACKGROUND AND OBJECTIVES: Diabetes mellitus (DM) is a risk factor for periodontal diseases and may exacerbate the progression of the pathogenesis of periodontitis. Advanced glycation end-products (AGEs) cause DM complications relative to levels of glycemic control and larger amounts accumulate in the periodontal tissues of patients with periodontitis and DM. In the present study, we investigated the effects of AGEs on the expression of inflammation-related factors in human gingival fibroblasts (HGFs) to elucidate the impact of AGEs on DM-associated periodontitis. MATERIAL AND METHODS: HGFs were cultured with or without AGEs. Cell viability was examined, and RNA and protein fractions were isolated from AGE-treated cells. The expression of interleukin (IL)-6, intercellular adhesion molecule-1 (ICAM-1), and the receptor for AGE (RAGE) was investigated using reverse transcription-polymerase chain reaction, quantitative real-time polymerase chain reaction and enzyme-linked immunosorbent assay, and reactive oxygen species activity was measured using a kit with 2',7'-dichlorofluorescin diacetate. Human monocytic cells (THP-1) labeled with a fluorescent reagent were co-cultured with HGFs treated with AGEs and IL-6 siRNA, and the adhesive activity of THP-1 cells to HGFs was assessed. The expression of IL-6 and ICAM-1 was examined when HGFs were pretreated with recombinant human IL-6, the siRNAs of RAGE and IL-6, and inhibitors of MAPK and NF-κB, and then cultured with and without AGEs. The phosphorylation of MAPK and NF-κB was assessed using western blotting. RESULTS: AGEs increased the mRNA and protein expressions of RAGE, IL-6, ICAM-1 and reactive oxygen species activity in HGFs, and promoted the adhesion of THP-1 cells to HGFs, but had no effect on cell viability until 72 hours. Recombinant human IL-6 increased ICAM-1 expression in HGFs, while the siRNAs of RAGE and IL-6 inhibited AGE-induced IL6 and ICAM1 mRNA expression, and IL-6 siRNA depressed AGE-induced THP-1 cell adhesion. AGEs increased the phosphorylation of p38 and ERK MAPKs, p65 NF-κB and IκBα, while inhibitors of p38, ERK MAPKs and NF-κB significantly decreased AGE-induced IL-6 and ICAM-1 expression. CONCLUSION: AGEs increase IL-6 and ICAM-1 expression via the RAGE, MAPK and NF-κB pathways in HGFs and may exacerbate the progression of the pathogenesis of periodontal diseases.

YKL-40 level in gingival crevicular fluid from patients with periodontitis and type 2 diabetes.

OBJECTIVE: YKL-40 is a chitin-binding glycoprotein, the level of which increases in inflammatory diseases, diabetes mellitus (DM), cardiovascular diseases, and tumors. Gingival crevicular fluid (GCF) contains many proteins and markers of periodontitis. The purpose of this study was to investigate YKL-40 level in GCF from patients with periodontitis and DM and the association between YKL-40 level and chronic periodontitis (CP) or DM. SUBJECTS AND METHODS: The subjects were 121 patients with DM, CP, DM and periodontitis (DM-P), and healthy subjects (H). GCF was collected using paper strips after the sites for GCF collection were clinically evaluated for probing depth (PD), gingival index (GI), and bleeding on probing (BOP). YKL-40 in GCF was identified by Western blotting, and its level was determined by ELISA. RESULTS: YKL-40 was contained in GCF samples from H, DM, CP, and DM-P sites, and its levels (amount and concentration) in CP and DM-P were significantly higher than those in H and DM. GCF YKL-40 level significantly correlated with PD and GI, and its level in BOP-positive sites was significantly higher than that in BOP-negative ones. CONCLUSIONS: GCF YKL-40 level was elevated in periodontitis, but not DM. YKL-40 in GCF may be an inflammatory marker for periodontitis.

[Surgery for thymomas and thymic carcinomas; treatment results in terms of WHO histologic typing, Masaoka staging system, and p53 expression].

Forty thymomas and thymic carcinomas were classified in terms of WHO histologic typing, Masaoka staging system, and p53 expression. In WHO histologic typing, type A, AB, B1, B2, B3, and C were 1, 10, 16, 5, 4, and 4 cases, respectively. In Masaoka staging system, I, II, III, and IV were 15, 9, 10, and 6 cases, respectively. Thirteen thymomas exhibited positive p53 expression and 27 did not. Type A and AB thymomas had more favorite prognosis than type B3 and C thymomas, and prognosis of type B1 and B2 was middle. Staging by the Masaoka system also correlated with survival rates. Patients who had p53-negative thymomas survived longer than those who had p53-positive thymomas. A treatment strategy for thymomas and thymic carcinomas should be made on the basis of WHO histologic typing, Masaoka staging system, and p53 expression.

[Prognostic factors for oral tongue carcinoma treated with intra-oral cone electron beam irradiation].

Twenty-five patients with squamous cell carcinoma of the oral tongue were treated with intra-oral cone electron beam irradiation (IOC) during the period from 1985 to 1995. We analyzed the prognostic factors for local control and complications. IOC was applied for T1 (16 cases) and T2 (9 cases) tumors. Hypofractionation was used for IOC (20, 10, or 8 Gy/fr, 1f/wk). The total dose delivered ranged from 40 Gy to 78 Gy. Radiation dose homogenization was done through calculation of the normalized total dose (NTD) for alpha/beta = 10 (tumor) and alpha/beta = 3 (late normal tissue). The two-year local control rates for T1 and T2 were 80.4% and 77.8%, respectively. The two-year local control rates for patients whose overall treatment time (OTT) was < or = 28 days (n = 16) was 100% vs. 41.7% for patients whose OTT was > 28 days (n = 9) (p = 0.002). Multivariate analysis was applied to identify possible prognostic factors for local control. OTT (p = 0.02) was the only variable that significantly influenced local control. The incidence of radiation ulcer was 33.3% (7/21). Significant indicators of ulceration were fraction size (>> 10 Gy) and NTD (alpha/beta = 3) (>> 130 Gy) (p < 0.05). These results indicate that prolonged OTT was the major reason for the failure of IOC radiotherapy to control local disease and that the relatively high rate of ulceration was due to large fraction size and high NTD (alpha/beta = 3).

Multi-fractionated wide-field radiation therapy for palliation of multiple symptomatic bone metastases from solid tumors.

PURPOSE: This was a pilot study to explore the toxicity and response of multi-fractionated wide-field radiation therapy (MF-WFRT) in patients with multiple symptomatic osseous metastases. PATIENTS AND METHODS: From February 1997 to April 1998, a total of nine patients (5 lung cancer, 3 breast cancer, 1 prostate cancer) were treated with MF-WFRT. The patients received 1.5 Gy per fraction (twice a day) to a total dose of 7 Gy in 3 days for the upper body and a total dose of 9 Gy in 3 days for the lower body. Ten treatments in nine patients were carried out with this technique (2 upper half-body, 5 lower half-body, 3 mid-body; one patient had both upper mid-body and lower-half body treatments). RESULTS: Pain relief was complete in two patients (20%) and eight (80%) achieved better than 50% pain relief. Seventy-five percent of pain responders achieved pain relief within one week of MF-WFRT. The pain relief was long-lasting and continued without need of reirradiation for 45% of the remainder of the patients' lives. The incidence of gastrointestinal complications was low, basically Grade 1-2 toxicity. Four patients (40%) experienced Grade 3-4 hematological toxicity. Hematological toxicity was treated with blood transfusion or G-CSF. General tolerance was excellent, and no pneumonitis or radiation-related deaths occurred. CONCLUSION: This treatment modality appears to be well tolerated and effective. The optimal indications, dose, and fractionation for MF-WFRT should be further explored in randomized studies.

Nucleic acid-binding properties and subcellular localization of the 3a protein of brome mosaic bromovirus.

Brome mosaic bromovirus (BMV) 3a protein is required for cell-to-cell movement of the virus in host plants. The BMV 3a protein (B3a) was produced in Escherichia coli using an expression vector. Gel retardation analysis and UV cross-linking experiments demonstrated that B3a bound single-stranded RNA cooperatively without sequence specificity. Binding competition analysis showed that B3a bound to single-stranded nucleic acids more strongly than to double-stranded nucleic acids. Deletion mutagenesis located a nucleic acid-binding domain to amino acids 189-242. Western blot analysis of fractionated proteins of BMV-infected barley using monoclonal antibodies against B3a indicated that B3a may interact with membrane materials and form complexes in the cytoplasm. Immunogold labelling of thin sections of infected barley tissues revealed that B3a was associated with plasmodesmata and cytoplasmic inclusions.

[Cure by THP-COP therapy in patients with perforated T-cell type malignant lymphoma of the jejunum].

A 52-year-old man had suffered abdominal pain from Dec. 15, 1992. On Jan. 1993, he was admitted to our hospital for a diagnosis of T-cell malignant lymphoma of stomach of diffuse large cell type by gastroendoscopical biopsy. On the following day, he underwent emergency an operation with a diagnosis of panperitonitis. A perforation site had been found at the jejunum 60 cm distant from the Treitz ligament. It was resected and sutured concomitant with omental patch. The pathological diagnosis was the same. After the operation, we started THP-COP therapy on Jan. 25, 1993. During the admission, he was given THP-COP therapy 6 times, and had a complete remission. He was discharged Feb. 26, 1994, and shows no evidence of disease at this writing.

[Effect of intraperitoneal therapy in gastric cancer with peritoneal dissemination].

Eighty gastric cancer patients with peritoneal dissemination were analyzed to evaluate the effect of intraperitoneal chemo- and/or immunotherapy on abdominal ascites. Among them, 19 were treated with intraperitoneal chemo- and/or immunotherapy, and 21.1% of them showed decreased ascites and better QOL. Patients treated with intraperitoneal administration with OK-432 showed good survival. Among patients with OK-432 administration, those receiving postoperative chemotherapy showed better survival than those given immunochemotherapy. Conversely, among patients with chemotherapy, those given postoperative immunochemotherapy showed better survival than those on chemotherapy. Patients with HLA-type I and III, and those with preoperative normal immune status showed good response when they received intraperitoneal therapy.

[Sjögren's syndrome complicated by thymoma].

A 43-year-old woman with a history of Sjögren's syndrome was admitted to our hospital because of dyspnea due to anemia. Her chest X ray film on admission revealed an anterior mediastinal tumor. We diagnosed this tumor as a thymoma based on a chest CT scan and magnetic resonance imaging. Because the latter indicated that the thymoma may have involved the mediastinal great vessels, preoperative chemotherapy with cisplatin, doxorubicin hydrochloride, vincristine sulfate and cyclophosphamide. Thereafter the tumor was successfully resected along with part of the right pleura. Thymoma as a complication of Sjögren's syndrome is rare. Here we also discuss the treatment of thymoma and the combination of these two diseases.

[A case of advanced gastric cancer with liver metastasis with no recurrence and long survival].

A 61-year-old male was admitted after detecting gastric lesion by gastrography in a medical health examination. The patient was diagnosed as Borrmann 2 advanced gastric cancer with remarkable intraperitoneal lymph node metastasis and liver tumor of lateral segment. Relative non-curative gastrectomy was performed with combined partial hepatectomy. The liver tumor measured 1.5 x 1.5 cm and was intraoperatively diagnosed as metastasis of gastric cancer. Mitomycin C 26 mg was given intravenously on the day of operation and 5-fluorouracil (5-FU) 150 mg/day orally since postoperative 14th day as adjuvant chemotherapy. The administration of 5-FU was continued for 5 years. As a result of such combination therapy, the patient still has had no recurrence 8 years following operation.

[Alteration of gamma-aminobutyric acid in streptozotocin-induced diabetic rat retina].

To elucidate possible alteration of gamma-aminobutyric acid (GABA) in the diabetic retina, the distribution and determination of GABA was analyzed in streptozotocin-induced diabetic and normal rats after electroretinogram (ERG) recording. Immunoreactivity of GABA was found in the inner nuclear layer, inner plexiform layer, and ganglion cell layer in normal and diabetic rats. In the inner nuclear layer, strong immunoreactivity of GABA was found in amacrine-like cells. In diabetic retinas, GABA immunoreactivity was higher than in normal retinas. The contents of GABA increased began 1 week after occurrence of diabetes mellitus, attained a maximum at 2 months, and maintained this amount for 5 months. The latencies of oscillatory potentials of ERG were prolonged beginning 1 month after occurrence. These results suggest that increase of GABA in the amacrine cells of diabetic retinas may be related with the abnormality of oscillatory potentials.

[A case of primary alveolar hypoventilation syndrome with a good response to nocturnal low-flow oxygen inhalation and negative pressure ventilation].

A 34-year-old female was admitted on June 25, 1990, for the evaluation of alveolar hypoventilation which worsened after her second delivery. She showed impairment of both hypercapnic and hypoxic ventilatory responses, and marked desaturation due to hypopnea and apnea during sleep. Although administration of methylxanthine and medroxyprogesterone was not very effective, after treatment with low flow oxygen, there was a marked decrease in the frequency and duration of desaturation during sleep and improvement of arterial daytime blood gases, which suggested the existence of hypoxic ventilatory depression in the pathophysiology of her nocturnal desaturation. Furthermore, the use of a negative pressure ventilator for 3 hours in the daytime for 10 days resulted in marked improvement of symptoms, arterial blood gases, respiratory muscle strength, and the frequency and duration of sleep desaturation. These findings suggest that both low flow oxygen therapy during sleep, and daytime negative pressure ventilation may be beneficial in patients with primary alveolar hypoventilation and central sleep apnea syndrome.

Dose equivalence for high-dose-rate to low-dose-rate intracavitary irradiation in the treatment of cancer of the uterine cervix.

By comparing the incidence of major radiation injury, we estimated doses clinically equivalent for high-dose-rate (HDR) to conventional low-dose-rate (LDR) intracavitary irradiation in patients with Stages IIb and IIIb cancer of the uterine cervix. We reviewed a total of 300 patients who were treated with external beam therapy to the pelvis (50 Gy in 5 weeks) followed either by low-dose-rate (253 patients) or high-dose-rate (47 patients) intracavitary treatment. The high-dose-rate intracavitary treatment was given 5 Gy per session to point A, 4 fractions in 2 weeks, with a total dose of 20 Gy. The low-dose-rate treatment was given with one or two application(s) delivering 11-52 Gy to the point A. The local control rates were similar in both groups. The incidence of major radiation injury requiring surgical intervention were 5.1% (13/253) and 4.3% (2/47) for low-dose-rate and high-dose-rate groups, respectively. The 4.3% incidence corresponded to 29.8 Gy with low-dose-rate irradiation, thus, it was concluded that the clinically equivalent dose for high-dose-rate irradiation was approximately 2/3 (20/29.8) of the dose used in low-dose-rate therapy.

A comparison of swine abdominal temperature distribution after heating with BSD-1000, an annular phased array system (APAS), and Thermotron RF-8.

External hyperthermia for deep-seated tumors is still a difficult problem. We compared the abdominal temperature distribution in a pig after heating with BSD-1000, an annular phased array system (APAS), and Thermotron RF-8. Thirty elastic tubes were inserted into the upper abdomen of the pig and a thermocouple was inserted into each of the tubes. After heating, these thermocouples were moved at 1-cm intervals and the three-dimensional temperature distribution was obtained. The temperature distribution after heating with APAS and Thermotron RF-8 was compared by using the distribution on the longitudinal center line of each sagittal plane at 1-cm intervals from the center. The temperature was much higher and its distribution more homogeneous with APAS than Thermotron RF-8.

[Radiotherapy for endobronchially invading recurrence of esophageal cancer after resective surgery].

Sixty-eight patients with endobronchially invading recurrence of esophageal cancer after resective surgery were treated with radiotherapy from 1966 to 1988. The mean interval between resective surgery and diagnosis of recurrence was 11.1 months, that was significantly shortened in a3 group. The dose of radiation for recurrence ranged from 2 to 70.3 Gy, with a mean dose of 42.6 Gy. The mean survival time after treatment of recurrence was 4.9 months. The dose of radiation was found to have a positive correlation with survival time. The cause of death was bleeding in 20 patients, and respiratory failure in 36. High dose of radiation was thought to induce high incidence of bleeding. The results indicated that external beam radiotherapy with conventional fractionation was not so much effective for the recurrence.

[Control of aftereffects due to intraoperative radiotherapy].

In cases of treating a pancreatic cancer, an aneurysm and thrombus of the abdominal aorta, necrosis of the vertebral body, and damage to the peripheral nerves can be caused by intraoperative radiotherapy (IORT). These aftereffects have been observed in tests conducted in experimental animal reported by Colorado State University. Similarly, our clinical experiences have led us to set safety guidelines to prevent damage due to IORT. The safety level of a radiation dose for IORT alone is limited to 30 Gy, and for IORT combined with external beam radiotherapy, from 20 Gy (IORT) plus 50 Gy (EBRT). Using these parameters, among 58 cases of a pancreatic tumor treated by IORT, only one case subsequently developed a pseudoaneurysm at the stump of splenic artery.

Carcinoma of the uterine cervix treated by irradiation alone. Results of treatment at the National Cancer Center, Tokyo.

Six hundred and twelve patients with previously untreated invasive carcinoma of the uterine cervix were treated by irradiation alone at the National Cancer Hospital from 1972 to 1983. The number of patients was 7, 39, 43, 127, 15, 319, 28 and 34 in stages IA, IB, IIA, IIB, IIIA, IIIB, IVA and IVB respectively. Low-dose-rate intracavitary irradiation with or without external irradiation was used in 383 patients, high-dose-rate intracavitary irradiation with or without external irradiation in 130, external irradiation alone in 98, and external irradiation combined with radon-222 seed implantation in one patient. Five-year-survival rates were 85, 65, 57, 41, 14 11% for stages IB, IIA, IIB, IIIB, IVA, and IVB respectively. The rate of complications was rather high in the present series, and so we have been investigating whether it is possible to reduce the dose. Low-dose-rate intracavitary irradiation has been replaced by high-dose-rate irradiation by using a remotely controlled afterloading system.

Local effect of hyperthermia for superficial and shallow-seated tumors.

In approximately seven years, 134 patients with 161 tumors were treated by hyperthermia combined with radiation or chemotherapy at our department. The primary tumors were breast cancer, head and neck cancer, and soft tissue tumors in most patients. Adenocarcinoma was the most frequent, followed by squamous cell carcinoma and soft tissue sarcoma. The local response rates for primary inoperable advanced, metastatic, and local recurrence of breast cancer were 88% (7/8), 50% (10/15), and 86% (18/21), respectively. The local response rate of 39 tumors of neck lymph nodes was 49% (19/39). A total of 26 tumors of bone and soft tissue were treated. Five tumors showed CR and six PR, for a total response rate of 42%. Among 20 patients with malignant melanoma, CR and PR were 25% (5/20) and 30% (6/20), respectively. The local response rate for all patients with superficial and shallow-seated tumors was 58% (94/161). In some tumors classified as showing NR, complete disappearance of tumor cells was demonstrated by a post-treatment histological examination. The efficacy of hyperthermia, when evaluated solely on the basis of tumor size, is likely to be underestimated.

[Stage IV renal cell carcinoma producing alpha-fetoprotein treated with multimodal treatment. A case report].

A 63 year old man, having right renal cell carcinoma which was found after initial symptoms of left supraclavicular tumor and gross hematuria, is presented. The points to note with this case are; (1) An unusual production of alpha-fetoprotein (AFP), (2) Being discovered at a very advanced stage and that the disease progressed rapidly (at initial presentation the patient had lymph nodes metastases from the paraaorta, mediastinum to the left supraclavicular region. Later the patient had a recurrent tumor in the left supraclavicular area and multiple cutaneous metastases), (3) Various modalities of treatment were given (right nephrectomy, abdominal lymph nodes dissection, resection of the supraclavicular and mediastinal lymph nodes metastases, hyperthermia combined with irradiation for the recurrent tumor in the supraclavicular region and systemically adoptive immunotherapy with tumor infiltrating lymphocytes). During this treatment course, the drift of serum AFP showed a good correlation. The possibility of AFP as a tumor marker of renal cell carcinoma in selected cases was presented and the suitability of the treatments performed to this patient was discussed.