Anti-Carcinogenic Effects of a Serine Protease Inhibitor (FOY-305) through the Suppression of Neutral Serine Protease Activity During Chemical Hepatocarcinogenesis in Rats

Anti-carcinogenic effect of a serine protease inhibitor, [N, N-dimethylcarbamoylmethyl 4-(4-guanidinobenzoyloxy)-phenylacetate] methanesulfate (FOY-305), was studied in rats with neutral serine protease during hepatocarcinogenesis induced by a single intraperitoneal administration of diethylnitrosamine (DEN) and following feeding of a diet containing 2-N-fluorenylacetamide (FAA) for 32 weeks. Oral administration of FOY-305 significantly suppressed development of -γ-glutamyl transpeptidase (γ-GTP)positive hyperplastic nodules, preneoplastic lesion, at the 8th week of DEN injection, and that of hepatocellular carcinoma (HCC) formation at the 32nd week. Neutral protease activity increased in the preneoplastic and neoplastic livers. The activities in the preneoplastic and tumor-bearing livers were much lower in FOY-305-treated group compared with those in control group. Neutral protease partially purified from neoplastic liver at the 32nd week was inhibited by FOY-305 in vitro. The data suggest that neutral protease plays a crucial role in the process of chemical hepatocarcinogenesis

Enrichment of Spermatogonial Stem Cells using Side Population in Teleost

Spermatogenesis originates from a small population of spermatogonial stem cells; this population can maintain continuous sperm production throughout the life of fish via self-renewal and differentiation. Despite their biological importance, spermatogonial stem cells are not thoroughly characterized because they are difficult to distinguish from their progeny 5 cells that become committed to differentiation. We previously established a novel technique for germ cell transplantation to identify spermatogonial stem cells based on their colonizing activity and their ability to initiate donor-derived gametogenesis in the rainbow trout (Oncorhynchus mykiss). Although spermatogonial stem cells can be retrospectively identified after transplantation, there is currently no technique to prospectively enrich for or purify spermatogonial stem cells. Here, we describe a method for spermatogonial stem-cell enrichment using side-population. With optimized Hoechst 33342 staining conditions, we successfully identified side-population cells among type A spermatogonia. Side-population cells were transcriptomically and morphologically distinct from non-side-population cells. To functionally determine whether the transplantable spermatogonial stem cells were enriched in the side-population fraction, we compared the colonization activity of side-population cells with that of non-side-population cells. Colonization efficiency was significantly higher with side-population cells than with non-side-population cells or with total type A spermatogonia. In addition, side-population cells could produce billions of sperm in recipient. These results indicated that transplantable spermatogonial stem cells were enriched in the side-population fraction. This method will provide biological information that may advance our understanding of spermatogonial stem 20 cells in teleosts. Additionally, this technique will increase the efficiency of germ-cell transplantation used in surrogate broodstock technology

Preliminary result of the relationship between the breathing frequency and dynamic body acceleration

March 5-6, 2009, Bangkok, ThailandIn this study, the relationship between the breathing frequency and the dynamic body acceleration (DBA) of one hatchery-reared loggerhead turtle Carretta carretta was examined using acceleration data loggers. Two acceleration data loggers (M190L-D2GT, W1000-3MPD3GT, Little Leonard, Japan) were attached on the lower-beak and carapace of a hatchery-reared loggerhead turtle, respectively. Breathing was successfully detected from the angle and depth of the beak-attached data logger and DBA, which has been used as an index of activity levels (Wilson et al., 2006), was calculated from the forward acceleration of the carapace-attached logger. There was a positive correlation between the DBA in the previous dive and the breathing frequency; the relationship was exponential. The result suggests that the number of breaths increased exponentially after a more active dive

Dermoscopic Features of a Black Hairy Tongue in 2 Japanese Patients

Dermoscopic features of a black hairy tongue have never been reported. Dermoscopy might be useful in speculating pathologic features of oral lesions. The objective was to identify additional dermoscopic criteria. Two Japanese patients who were clinically given a diagnosis of “black hairy tongue” were evaluated for dermoscopic features. We have shown characteristic dermoscopic features of brownish hair-like elongation of filiform papillae with whitish lingual papillae. Dermoscopic examination seemed useful as an adjunct to the diagnosis of this benign disorder of the tongue, demonstrating exact changes in shape and color of filiform papillae. It might also be helpful in more objective observation of the therapeutic efficacy

Two Cases of Severe Degeneration of the Macula Following Vitrectomy with Indocyanine Green-Assisted Internal Limiting Membrane Peeling for Idiopathic Macular Hole

We describe three eyes of two cases of severe degeneration of the macula following vitrectomy with indocyanine green-assisted internal limiting membrane peeling for idiopathic macular hole. We need to remember the possibility of these complications and have to select the procedures that are safest to use for macular hole surgery

Late-onset peritoneal recurrence of advanced gastric cancer 20 years after primary resection

Late onset of peritoneal recurrence of gastric cancer more than 10 years after surgery is extremely rare, and only three cases have been reported. We present the case of a 61-year-old man who was diagnosed finally with peritoneal recurrence of gastric cancer 20 years after primary curative resection. As a result of small-bowel obstruction caused by peritoneal recurrence, diverting ileostomy with partial ileal resection was performed. The resected specimen revealed tubular adenocarcinoma that resembled the primary gastric cancer. The clinical course after the second operation was unfavorable and systemic chemotherapy had no effect. He died at 62 years of age, 21 years and 7 months after initial gastrectomy. Immunohistochemical analysis using antibodies against proliferating cell nuclear antigen (PCNA), Ki-67, and p53 was performed to investigate the phenotype of primary and recurrence cancer. Protein expression of proliferation markers such as PCNA and Ki-67 was down-regulated, but p53 was overexpressed at the site of recurrence. These data suggest that late peritoneal recurrence has a low proliferation rate and is resistant to chemoradiotherapy. In conclusion, we present late onset of peritoneal recurrence of gastric cancer more than 20 years after primary surgery, and speculate on the mechanism of late-onset recurrence in our case

Proliferation and cell death of human glioblastoma cells after carbon-ion beam exposure: Morphologic and morphometric analyses

Histological analyses of glioblastoma cells after carbon-ion exposure are still limited and ultrastructural characteristics have not been investigated in detail. Here we report the results of morphological and morphometric analyses of a human glioblastoma cell line, CGNH-89, after ionizing radiation to characterize the effect of a carbon-beam on glioblastoma cells. Using CGNH-89 cells exposed to 0–10 Gy of X-ray (140kVp) or carbon-ions (18.3 MeV/nucleon, LET = 108 keV/μm), we performed conventional histology and immunocytochemistry with MIB-1 antibody, transmission electron microscopy, and computer-assisted, nuclear size measurements. CGNH-89 cells with a G to A transition in codon 280 in exon 8 of the TP53 gene had nuclei with pleomorphism, marked nuclear atypia and brisk mitotic activity. After carbon-ion and X-ray exposure, living cells showed decreased cell number, nuclear condensation, increased atypical mitotic figures, and a tendency of cytoplasmic enlargement at the level of light microscopy. The deviation of the nuclear area size increased during 48 hours after irradiation, while the small cell fraction increased in 336 hours. In glioblastoma cells of the control, 5 Gy carbon-beam, and 10 Gy carbon-beam, and MIB-1 labeling index decreased in 24 hours (12%, 11%, 7%, respectively) but increased in 48 hours (10%, 20%, 21%, respectively). Ultrastructurally, cellular enlargement seemed to depend on vacuolation, swelling of mitochondria, and increase of cellular organelles, such as the cytoskeleton and secondary lysosome. We could not observe apoptotic bodies in the CGNH-89 cells under any conditions. We conclude that carbon-ion irradiation induced cell death and senescence in a glioblastoma cell line with mutant TP53. Our results indicated that the increase of large cells with enlarged and bizarre nuclei, swollen mitochondria, and secondary lysosome occurred in glioblastoma cells after carbon-beam exposure.学位記番号：医博甲1096, 学位の種類：博士（医）, 学位授与年月日：平成20年3月25

Tumor-specific exon creation of the HELLS/SMARCA6 gene in non-small cell lung cancer

In an attempt to identify tumor suppressor genes on chromosome 10 in non-small cell lung cancers, we isolated 10 types of splicing variants of the HELLS/ SMARCA6 gene transcripts. HELLS/SMARCA6 is a novel member of SNF2 family, which is implicated in cellular function like chromatin remodeling. Variant 1 was an alternatively spliced isoform containing an insertion of a 44-ntd intronic sequence between exons 3 and 4, giving rise to a premature termination of translation. The expression of the variant 1 was detected exclusively in the lung cancer specimens (11 of 43 cases, 26%), but was not detected in corresponding normal tissues. D10S520 marker in the proximity of the HELLS/SMARCA6 gene showed prevalent allelic loss (41%) as compared with flanking markers (25-31%). These results suggest that loss of function of HELLS/SMARCA6 by allelic loss and aberrant proteins by tumor-specific exon creation may result in epigenetic deregulation, leading the lung cells to malignancy or its progression