The degree of DNA instability as determined by immunohistochemical staining with anti-singlestranded DNA antibody after acid hydrolysis (the DNAinstability test) was used as a marker of malignancy. The test was applied to tissues of oral leukoplakia assessed histopathologically as hyperplasia (38 cases), mild (12 cases), moderate (11 cases) and severe (8 cases) dysplasia, and invasive squamous cell carcinoma (SCC, 20 cases). Tissues were subjected to immunohistochemical staining for proliferating cell nuclear antigen (PCNA), p53, DNA-fragmentation factor 45 (DFF45), analysis of various AgNORs parameters, and triple immunostaining for vascular endothelial growth factor (VEGF), CD34, and PCNA. The DNA instability test was positive in 20 (100%) SCC cases, 8 (100%) severe dysplasia cases, 8 (72.7%) moderate dysplasia cases, 6 (50.0%) mild dysplasia cases, and 9 (23.7%) hyperplasia cases, indicating malignancy. The proportion of lesions positive for PCNA, p53, DFF45, and values of AgNORs parameters steadily increased from hyperplasia to mild, moderate and severe dysplasia, and SCC, especially in those showing positive DNA instability test, indicative of malignancy. Based on these results, 44.9% of leukoplakia were malignant tissues, namely carcinoma in situ. The proportion of PCNA-positive vascular endothelial cells in the vicinity of VEGF-positive epithelial lesion was significantly higher than that of negative DNA instability lesions, as revealed by immunohistochemical triple staining for VEGF, CD34, and PCNA. Our results suggest that increased DNA instability, enhanced proliferative activity, p53 mutation, and induction of DFF45 and VEGF may allow cancer cell proliferation, enhance their survival by escaping apoptosis, and provide abundant nutrients during early-stage carcinogenesis of oral leukoplakia

Growth hormone-releasing hormone (GHRH) is a well-known hypothalamic hormone that stimulates the synthesis and release of growth hormone (GH) as well as the proliferation of GH-producing cells in the anterior pituitary gland. Recent reports have shown GHRH synthesis in pituitary somatotroph adenomas, but GHRH immunoreactivity has not been demonstrated in the previous studies. In order to confirm the role of locally generated GHRH for the progression of somatotroph adenomas, we investigated the expression of GHRH in 25 pituitary somatotroph adenomas immunohistochemically using both conventional avidin-biotin-complex (ABC) method and novel catalyzed signal amplified (CSA) system.In addition, we investigated the expression of GHRH mRNA and GHRH receptor mRNA with in situ hybridization (ISH) using CSA system. The weak immunopositivity of GHRH were observed in only 2 adenomas (8.0%) out of 25 somatotroph adenomas using ABC method. In contrast, 15 adenomas (60.0%) out of 25 somatotroph adenomas were immunopositive for GHRH revealed by CSA system. The expression of GHRH mRNA was confirmed using CSA-ISH system in 13 adenomas (72.2%) out of 18 somatotroph adenomas. In 11 adenomas (61.1%) out of 18 somatotroph adenomas, the expression of GHRH receptor mRNA was demonstrated using CSA-ISH system. This is a first report that clarified histopathologically GHRH production in pituitary somatotroph adenomas. The demonstration of GHRH and its receptor expression is meaningful in clarifying the autocrinc or paracrine regulation of GHRH in GH production and progression of pituitary somatotroph adenomas. P-I-38Age-related changes of the microvasculature in the median eminence of rats. We have previously demonstrated that astroglial elements in the median eminence of aged rats (24 month-old rats), especially in the perivascular region in its outer layer, increased significantly using immunocytochemistry and computer-assisted analyzing system. According to aging. astrocytes in the perivascular region exhibited more intimate contact with the basement membrane of the perivascular space in the portal vessels compared to control groups (6 week-old rats). Therefore, age-related changes of astroglial elements are considered to affect on the microvasculature of the median eminence. In the present study, to clarify the effect of aging on the neuroendocrine system, the changes of microvasculature of the median eminence was investigated. The hypophyseal vascular plexus of aged and control rats were prepared for the injection of acrylic resin into the ascending aorta and examined with scanning electron microscope. In the saggital view, the plexuses of median eminence in the aged groups became thicker at the dorso-ventral level but these structures seemed to be not prominent change compared to those of control groups at the rostro-caudal level. The microvasculature of their capillaries increased in the diameter and exhibited to be more complex structure and they are considered to be sinusoid-like structure. In the anterior and posterior lobes, th...

The guideline issued by the Fisheries Agency prohibits fishing-boats from evacuating to the offing. While the present fishing insurance almost insure fishing-boats hit by tsunami, it doesn't insure life during suspension at all, the fishermen bear big burden economically. Therefore many fishermen evacuated fishing-boats to the offing to protect them, but we find some problem fishermen don't have enough knowledge of tsunami. In this study, we practice the survey of fishermen's consciousness and actual fishing-boat's evacuation, and we build rules of fishing-boat's evacuation and communication method, to promote fishermen's correct knowledge of tsunami and to evade evacuation on dangerous situation.

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