Ryo Konno's research while affiliated with Tohoku Medical and Pharmaceutical University and other places

Interleukin-18 (IL-18), a cytokine produced by macrophages, is capable of inducing T-lymphocyte synthesis of interferon-gamma (IFN-gamma). In this study, for the first time, the serum concentration of IL-18 and its significance as a prognostic indicator was evaluated in patients with epithelial ovarian cancer. The serum IL-18 level was measured by an enzyme-linked immunosorbent assay (ELISA) in 69 patients with epithelial ovarian cancer and 8 healthy controls. Relationships between the IL-18 level and clinicopathological features were examined by univariate and multivariate analyses. The median serum IL-18 level in the ovarian cancer patients was 229.6 pg/ml, and the level was significantly elevated compared with that in the normal controls (151.3 pg/ml; P < 0.01). No significant correlations were detected between the IL-18 level and stage or histology (P = 0.08 and P = 0.12, respectively). On univariate analysis, overall survival was shown to be affected by IL-18 serum levels. However, multivariate analysis failed to demonstrate an independent prognostic significance for IL-18 serum levels, while confirming the role of previously established prognostic variables, such as performance status, stage, and residual tumor. This study showed that IL-18 serum levels were elevated in ovarian cancer patients and were correlated with overall survival, although they were shown not to be an independent prognostic factor.

Objective: In situ metabolism and synthesis of oestrogens are considered to play important roles in the pathogenesis and development of human endometrial endometrioid adenocarcinoma. Approximately 3-5% of patients with these neoplasms are under age 40, some of whom have been treated with progestogen alone as a primary therapy for both atypical endometrial hyperplasia and adenocarcinoma in order to preserve their fertility. Medroxyprogesterone acetate (MPA) has been used extensively in the treatment of both breast and endometrial disorders as an endocrine therapy. However, details of the alterations of in situ oestrogen metabolism following progestogen treatment have yet to be fully elucidated. Design, patients and measurements: In this study we examined the immunolocalization of 17beta-hydroxysteroid dehydrogenase (17beta-HSD) types 1 and 2, oestrogen receptor (ER), progesterone receptor (PR)A + PRB, PRB, and Ki67 in progestogen-treated endometrial endometrioid adenocarcinoma (16 cases). We compared our findings both prior to and following treatment. These findings were then correlated with the treatment outcome of individual patients in order to elucidate factors associated with the response to treatment. Results: 17beta-HSD type 2 immunoreactivity was detected in 8/16 cases examined, whereas 17beta-HSD type 1 immunoreactivity was undetected in all cases examined. 17beta-HSD type 2 positive immunostaining, PRA + PRB labelling index (LI), and PRB/PRA + PRB ratio were all significantly higher in cases responding to the treatment than in those not responding. There were no significant correlations between responsive and nonresponsive cases for positive 17beta-HSD type 1 immunostaining, Ki67 LI, ER LI and age. There were no significant differences in the positive immunostaining for 17beta-HSD types 1 and 2, Ki67 LI, ER LI, PRA + PRB LI, age and PRB/PRA + PRB ratio between specimens taken prior to and following progestogen treatment. Conclusion: These results suggest that in situ abundance of 17beta-HSD type 2 and PR, especially PRB, can predict the possible response of patients with endometrial carcinoma to progestogen treatment.

Laparoscopic surgery has inherent restrictions with respect to the operative field of view and the range of surgical manipulation. Of the two procedures which secure sufficient operative space, the operative view of the gasless method is inferior to that of a pneumoperitoneum. In order to gain greater surgical visualization in gynecological gasless laparoscopy, the authors devised the cul-de-sac packing method employing a metreurynter, an instrument familiar to obstetricians in Japan. A metreurynter was lead into the cul-de-sac, and was inflated with saline, which resulted in the adnexae being raised up. This method was performed in three patients whose preoperative diagnoses were unknown infertility, ovarian cyst, and ectopic pregnancy, respectively. In all cases this method was able to keep the bowels out of the cul-de-sac space. In the first case, we were able to perform a tubal patency test under tension-free conditions, while at the same time bilateral tubal information could be obtained in a single view. In the latter two cases the adnexal lesions were maintained at an inspectional position throughout the operation without the necessity of being held by forceps to prevent them from falling down into the cul-de-sac space. No complications occurred in our three cases. This method will not be useful for patients whose cul-de-sac space is closed due to adhesions. However, except in such cases, this technique supplies a good operative view while being simple, safe, and inexpensive. Furthermore, this method supports gentler and less traumatic manipulation throughout the operation.

Intratumoral metabolism and synthesis of estrogens are consid- ered to play very important roles in the pathogenesis and development of human endometrial adenocarcinoma. The 17b-hydroxysteroid de- hydrogenase (17b-HSD) isozymes catalyze the interconversion of es- tradiol (E2) and estrone and thereby serve to modulate the tissue levels of bioactive E2. To elucidate the possible involvement of this enzyme in human endometrial carcinoma, we first examined the expression of 17b-HSD type 1 and type 2 in 20 normal cycling human endometria, 36 endometrial hyperplasia, and 46 endometrial endo- metrioid adenocarcinoma using immunohistochemistry, and we then studied immunoreactivity of 17b-HSD type 2 using immunoblotting analyses, the activity of 17b-HSD type 1 and type 2 using thin-layer chromatography and their expression using RT-PCR in endometrial endometrioid adenocarcinoma. We correlated these findings with var- ious clinicopathological parameters to examine the biological signif- icance of 17b-HSDs in human endometrial disorders. 17b-HSD type 2 immunoreactivity in normal endometrium was present in all cases of secretory phase (n 5 14), but not in any endometrial mucosa of proliferative phase (n 5 6). In addition, 17b-HSD type 2 immunore- activity was detected in 27 of 36 (75%) endometrial hyperplasia and 17 of 46 (37%) carcinoma cases. 17b-HSD type 1 immunoreactivity was not detected in all the cases examined. In both endometrial hyperplasia and carcinoma cases there were significant positive cor- relations between 17b-HSD type 2 and progesterone receptor labeling index (LI). In carcinoma cases, a significant inverse correlation was detected between 17b-HSD type 2 immunoreactivity and age. In ad- dition, 17b-HSD type 2 immunoreactivity was also correlated with 17b-HSD type 2 enzymatic activity, and semiquantitative analyses of 17b-HSD type 2 messenger RNA. No significant correlations were detected between 17b-HSD type 2 and estrogen receptor LI, Ki67 LI, amount of aromatase messenger RNA or histological grade. These data indicated that the expression of 17b-HSD type 2 in hyperplastic and/or neoplastic endometrium may represent altered cellular fea- tures through hyperplastic and neoplastic transformation. However, 17b-HSD type 2 may also play some protective and/or suppressive roles toward unopposed estrogenic effects through inactivating E2 in situ, especially in premenopausal patients. (J Clin Endocrinol Metab 86: 3436 -3443, 2001)

Retinoids have recently been proposed to modulate estrogenic actions in various sex steroid-dependent neoplasms, but little has been studied in human endometrial disorders. Therefore, in this study, we first examined the immunolocalization of retinoic acid receptor alpha, beta, and gamma, and retinoid X receptor (RXR) alpha, beta, and gamma in 20 normal cycling human endometria, 34 endometrial hyperplasia, and 46 endometrioid endometrial adenocarcinomas. We then correlated these findings with other clinicopathological parameters, especially in the correlation between retinoid receptor subtypes and the status of steroid hormone receptors, 17 beta-hydroxysteroid dehydrogenase (17 beta-HSD) and aromatase. We also then examined the effects of retinoic acid on the expression of 17 beta-HSD type 2 in cell lines derived from endometrial carcinoma using Northern blotting analysis to examine the possible roles of retinoids in in situ endometrial estrogen metabolism. Among these six retinoid receptors examined, RXR gamma immunoreactivity was exclusively detected in the epithelial cells of the secretory phase endometrium but not of the proliferative phase, which was well correlated with 17 beta-HSD type 2 immunolocalization. However, in endometrial hyperplasia, RXR gamma was not correlated with 17 beta-HSD type 2. In endometrioid endometrial adenocarcinoma, there was a statistically significant correlation between 17 beta-HSD type 2 immunoreactivity and RXR gamma labeling index (LI) (P < 0.001) and between RXR gamma LI and progesterone receptor LI (r = 0.501, P = 0.003). A significant inverse correlation was also detected between RXR gamma LI and patient age (r = 0.449, P = 0.015). No statistically significant correlation was obtained between LIs of receptors and other clinicopathological parameters including the status of intratumoral aromatase examined by immunohistochemistry. In the endometrial carcinoma cell line, RL95-2, retinoic acid markedly increased the level of 17 beta-HSD type 2 messenger RNA in a time- and dose-dependent manner. These results all suggest that retinoic acids may be involved in modulation of in situ estrogen metabolism in both normal and neoplastic human endometrium possibly through RXR gamma by stimulating the expression of 17 beta-HSD type 2.

Objective: Report of a remnant tubal pregnancy after cloacal malformation repair. Design: Case report. Setting: A university hospital. Patient(s): A woman with cloacal malformation repair. Intervention(s): Laparoscopic surgery. Main Outcome Measure(s): Remnant tubal pregnancy. Result(s): Laparoscopic right salpingectomy. Conclusion(s): In this patient, ectopic pregnancy was caused by contralateral sperm transmigration to the remnant right tube, which was not connected with the left uterus. (C)2001 by American Society for Reproductive Medicine.

A questionnaire survey was administered by Tohoku University hospital to investigate the current status of gynecological laparoscopic operations in teaching hospitals in which residents were trained as obstetricians and gynecologists. Thirty-eight senior doctors in 18 hospitals were sent a postal questionnaire. The response rates of facilities and doctors were 94.4% and 76.3% respectively. Diagnostic and operative laparoscopy was performed in all of the hospitals surveyed, however only a few hospitals performed more than 100 laparoscopies per year, and many surgeons did not perform advanced laparoscopic operations including hysterectomy, myomectomy and lymphadenectomy. Half of the doctors performed minilaparotomy, which is a so-called "open" technique. The standard surgical style involved the use of a carbon dioxide (CO2) pneumoperitoneum and an endoscope 10 mm in diameter. Unipolar endocoagulating instruments were generally used for hemostasis and incision. Most of the doctors surveyed thought that operative laparoscopy would become a necessary procedure in gynecological field, and half of them hoped to receive training in the technique. Instruction of expert endoscopic surgeons is necessary, especially in teaching hospitals, for the spread of safe and advanced laparoscopic surgery. An educational and training system for established surgeons as well as for young residents is urgently needed in Japan.

Our objective is to investigate the involvement of granule-mediated apoptosis in the cyclic changes of the endometrium. We demonstrated the localization of CD56, perforin, granzyme B and caspase-3 in the endometrium by immunohistochemistry. We also confirmed the localization of perforin by immuno-electron microscopy, and demonstrated apoptosis in endometrial glandular cells by TdT-mediated dUTP-biotin nick end labeling (TUNEL) and electron microscopy. Uterine CD56-positive natural killer (NK) cells expressed perforin and granzyme B in its cytoplasm. Uterine NK cells increased significantly in the endometrial stroma during the secretory phase, and peaked during the late secretory phase. These cells started decreasing in number during the menstrual period. In endometrial glandular cells, caspase-3 and TUNEL-positive cells increased significantly from the late secretory phase, with apoptosis reaching a peak during the menstrual period. Using electron microscopy, we observed uterine NK cells with chromatin rich, segmented nuclei and intracytoplasmic granules in the stroma obtained from late secretory phase endometria. These cells extended projections to the lining of endometrial glandular cells and attached to form a cell-to-cell contact. In addition, nuclear chromatin was observed to have already cohered and small cytoplasmic organelles were beginning to disappear, suggesting that these endometrial glandular cells were undergoing apoptosis. Utilizing immuno-electron microscopy, intracytoplasmic granules in uterine NK cells were stained with anti-perforin antibody. The findings of this study suggest that granule-mediated apoptosis in endometrial glandular cells induced by NK cells expressing perforin and granzyme B may be associated with the onset of menstruation.

Human epithelial ovarian carcinoma is well-known as a sex steroid-dependent neoplasm, but the possible biological significance of progesterone receptor (PR) in this cancer remains controversial. Recently, two isoforms of human PR, PRA and PRB, have been characterized and different functional characteristics have been reported for these two isoforms. We therefore examined immunohistochemistry (107 cases) and reverse transcription-polymerase chain reaction (RT-PCR) (16 cases) for PRA, PRB, and oestrogen receptor-a (ER-a). Labeling indices (LI) for PRA and PRB were 2.4 and 43.6, respectively, and the difference was statistically significant. PRB LI, but not PRA LI, as well as performance status, stage, and residual tumour turned out to be independent prognostic factors following multivariate analysis. There was also a significant correlation between ER-a LI and PRB LI (r = 0.595, P < 0.0001), suggestive of a possible interaction between these two receptors. RT-PCR also detected the expression of PR isoform transcripts in the same pattern as was observed with immunohistochemistry. Results of these studies indicate that PRA and PRB both mediate distinct pathways of progesterone action in ovarian carcinoma. Moreover, it is important to examine PRB LI as a prognostic factor in the cases of human epithelial ovarian carcinoma.

Estrogens play important roles in the pathogenesis of the great majority of endometrial endometrioid adenocarcinoma. Recently, a novel estrogen receptor (ER), ER beta, has been characterized, but little is known about the status of ER beta in endometrial carcinoma. We therefore examined expression of both ER alpha and ER beta in 45 cases of endometrioid endometrial adenocarcinoma using mRNA in situ hybridization, reverse transcription and polymerase chain reaction (RT-PCR), and immunohistochemistry. We also correlated the findings with various clinicopathologic parameters in these cases to examine their possible biologic significance. Accumulation of mRNA hybridization signals for both ER alpha and ER beta was detected predominantly in the cytoplasm of carcinoma cells, and to a lesser extent in some stromal cells. ER beta mRNA was detected in 16/45 cases (35.6%), and ER alpha mRNA hybridization signals were detected in 36/45 cases (80.0%). Among the 16 ER beta positive cases, 15 cases also had ER alpha mRNA hybridization signals. In the cases that expressed both ER alpha and ER beta, ER alpha mRNA hybridization signals were more widely distributed than ER beta mRNA. In 21 cases, carcinoma cells had ER alpha mRNA hybridization signals but not ER beta mRNA. There was a statistically significant positive correlation between the results of mRNA in situ hybridization and semiquantitative RT-PCR or immunohistochemistry for both ER alpha and ER beta. There were no significant correlations between ER beta mRNA expression and PR labeling index, Ki67 LI, age, or histologic grade. The results from our study indicate that ER beta is coexpressed with ER alpha, and that the estrogenic effects occur predominantly through ER alpha in endometrial carcinomas.