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Biomedical subjects

H Katabuchi

Publications and source records attributed to H Katabuchi.

At least 37 records · Page 2Linked to original sources

Establishment of a new cell line, OKT1, from small cell carcinoma secreting ectopic ACTH of the uterine cervix.

OBJECTIVE: Small cell carcinoma of the uterine cervix is rare and represents a unique entity among gynecological tumors. It sometimes demonstrates neuroendocrine differentiation, including adrenocorticotropin (ACTH) secretion. In this study, we established a new cell line, OKT1, from a case of carcinoma secreting ectopic ACTH without Cushing's syndrome and determined the character of the cell line. METHODS: OKT1 was established from OKT tumor cells, derived from a biopsy specimen of small cell cervical carcinoma, and serially heterotransplanted into nude mice. To characterize OKT1, the cell morphology, growth properties, immunohistochemical properties, hormone- and tumor-associated antigen secretion, tumorigenic potential, DNA profile, and chromosomal alteration were studied. RESULTS: The population doubling time of OKT1 was approximately 27 h. The cytological properties of OKT1, including DNA ploidy pattern, were similar to those of the primary tumor. Neuroendocrine differentiation was shown in the OKT1 cells by the positive immunocytochemical staining of neuron-specific enolase (NSE) and the presence of NSE and ACTH in the culture media. The xenograft of 1 x 10(8) OKT1 cells into nude mice yielded tumor mass. Furthermore, OKT1 demonstrated HPV type 18 and absence of a p53 gene mutation from exons 5 through 8. CONCLUSION: To our knowledge, OKT1 is the first cell line established from small cell cervical carcinoma with ACTH secretion.

Adrenocorticotropic Hormone↗

Carcinoma of the uterine cervix. High-resolution turbo spin-echo MR imaging with contrast-enhanced dynamic scanning and T2-weighting.

PURPOSE: To compare high-resolution contrast-enhanced (Gd-DTPA) dynamic MR imaging with T2-weighted turbo spin-echo (TSE) imaging in the evaluation of uterine cervical carcinoma. MATERIAL AND METHODS: Thirty-two patients with cervical carcinoma underwent MR imaging on a 1.5 T superconductive unit to have the extension of the disease assessed before treatment. A phased-array coil was used in all patients. In 25 patients, surgical confirmation of the diagnosis was obtained after imaging. Radiation therapy was selected for the remaining 7 patients with advanced carcinoma. Qualitative and quantitative image analyses were also performed. RESULTS: The cervical carcinomas showed maximum contrast in the cervical stroma and myometrium in the early dynamic phase. The tumor/cervical-stroma contrast in the early dynamic phase obtained with the T1-weighted TSE technique (contrast-to-noise ratio 22.6) was significantly higher than that obtained in T2-weighted TSE imaging (contrast-to-noise ratio 4.3). In the evaluation of parametrial invasion, the accuracy of T2-weighted imaging was 71.8% and contrast-enhanced dynamic imaging 81.2%. CONCLUSION: High-resolution contrast-enhanced (Gd-DTPA) dynamic MR imaging in cervical cancer offers improved tumor/cervical-stroma contrast and provides useful information on parametrial invasion.

Adult↗

Micropapillary serous carcinoma of the ovary: an immunohistochemical and mutational analysis of p53.

Micropapillary serous carcinoma (MPSC) has recently been described as a distinct ovarian neoplasm that shares histologic features with both serous borderline tumors (SBTs) and typical serous carcinomas of the ovary. To further define the relationship of MPSC to these two neoplasms, we evaluated all three tumor types for expression of the p53 protein and p53 gene mutations. The majority of MPSCs demonstrated positive, but only moderately intense, p53 immunostaining in >50% of the cells, whereas SBTs showed very weak staining in a small number of cells. In contrast, the majority of serous carcinomas displayed diffuse, very intense staining and those that did not stain completely lacked any staining for p53. This pattern of p53 immunostaining in MPSCs can be distinguished from the pattern observed in SBTs and in serous carcinomas. Both the MPSCs and the SBTs lacked p53 mutations in the cases analyzed, whereas all immunopositive serous carcinomas were found to have mutations in p53. In addition, one of the immunonegative cases of serous carcinoma had a frameshift mutation resulting in a truncated protein, providing a likely explanation for the lack of detectable p53 protein. These findings provide support for classifying MPSC as a distinct neoplasm of the ovary and suggest that increased expression of wild-type p53 may play a role in its pathogenesis.

Cystadenocarcinoma, Papillary↗

Distribution and cytological properties of macrophages in human Fallopian tubes.

The macrophages in human Fallopian tubes of women in the reproductive and postmenopausal periods were examined with a focus on their morphological properties by immunohistochemical staining and transmission electron microscopy. The fine structure of the smooth muscle cells in the Fallopian tubes was also investigated during the reproductive period. For immunohistochemical staining, we used two monoclonal antibodies that were specific for human macrophages, namely PM-1K and PM-2K. PM-1K recognizes human monocytes/ macrophages corresponding to CD68, and PM-2K recognizes tissue macrophages. PM-1K-positive cells were always present and their numbers increased significantly during the menstrual and early to mid-secretory phases. In contrast, relative numbers of PM-2K-positive cells were small throughout the menstrual cycle. In the postmenopausal period, few PM-1K-positive cells were detected, but PM-2K-positive cells remained. The macrophages during the secretory phase in the endosalpingeal stroma had well-developed intracytoplasmic organelles, but relatively few cytoplasmic vacuoles and granules. In the same phase, many cells of the monocyte/macrophage lineage appeared in the vascular lumen of the endosalpingeal stroma. The macrophages during the menstrual phase had well-developed intracellular organelles, with cytoplasmic vacuoles and granules of various sizes and configurations. During the late secretory phase, just prior to menstruation, the smooth muscle cells contained few cytoplasmic filaments but electron-lucent or electron-dense lysosome-like bodies were seen. These findings suggested the presence of macrophages in human Fallopian tubes. It is possible that such macrophages might be involved in the physiological functions of the tubes during the reproductive period and moreover that they might participate in the reconstruction of the muscle layer of the tubes.

Adult↗

Spatio-temporal changes of prolyl 4-hydroxylase in granulosa cells during ovulation in eCG-hCG-treated immature rat ovaries.

The immunolocalization of prolyl 4-hydroxylase (PHase), a key enzyme of collagen synthesis, and the effects of anti-progesterone RU486 on PHase during the ovulatory process in eCG-hCG-treated immature rat ovaries were studied to investigate the mechanisms of tissue repair in follicle walls after follicular rupture. Immunolocalization of PHase was studied using an anti-rat PHase subunit monoclonal antibody, and the amount of immunoreactive PHase was measured by enzymeimmunoassay. No obvious immunolocalization of PHase was observed in theca cells throughout the ovulatory process except just after follicular rupture. In contrast, in granulosa cells, PHase was first observed at 9 h after the hCG injection, and the staining intensity apparently increased from 9 to 15 h, especially around the apex of preovulatory follicles and the orifice of ruptured follicles. Consistent with these observations, PHase concentration in granulosa cells isolated from the ovaries significantly increased by 9 h (0.45 +/- 0.03 pg per cell), and reached a peak at 15 h (0.66 +/- 0.06 pg per cell) after the hCG injection. This peak was inhibited when 20 mg RU486 kg-1 was administered at 8 h (0.46 +/- 0.05 pg per cell), and the RU486-inhibited PHase concentration was recovered by the concomitant administration of 10 mg progesterone kg-1 (0.65 +/- 0.02 pg per cell). The results suggest that PHase expressed in granulosa cells may play an important role in the repair of ruptured follicle walls, via progesterone-dependent PHase production.

Analysis of Variance↗

Adenoma malignum: MR imaging and pathologic study.

PURPOSE: To evaluate the clinical, pathologic, and magnetic resonance (MR) imaging findings in adenoma malignum, a rare variant of uterine cervical adenocarcinoma. MATERIALS AND METHODS: Medical records of all patients (n = 7) with adenoma malignum of the uterine cervix diagnosed pathologically between 1988 and 1996 were retrospectively reviewed. Unenhanced T1-weighted and T2-weighted images and gadolinium-enhanced T1-weighted MR images were evaluated, and findings were correlated with gross pathologic and microscopic features. RESULTS: In five of seven patients, enlargement of the cervix was seen. All lesions were detected as multiple cystic lesions that extended from the endocervical gland to the deep stroma of the cervix. They appeared isointense (n = 5) or slightly hyperintense (n = 2) relative to the uterus on T1-weighted images and markedly hyperintense relative to the uterus on T2-weighted images. Solid portions of variable size were seen between cystic lesions, and both the multiple cystic component and the solid portion were most apparent on the gadolinium-enhanced T1-weighted images. Microscopic parametrial invasion was seen in two patients but was not detected at MR imaging. CONCLUSION: Adenoma malignum was depicted on MR images as a multicystic mass with solid portions located in the deep cervical stroma. Gadolinium enhancement helped identify the solid portion of the tumor.

Adenocarcinoma↗

Distribution and fine structure of macrophages in the human ovary during the menstrual cycle, pregnancy and menopause.

We evaluated the distribution and ultrastructural characteristics of macrophages in the ovaries of women of reproductive ages, during pregnancy, and after menopause, by immunohistochemistry and transmission electron microscopy. Macrophages appeared around the ovarian follicle with its development. Their organelles were poorly developed, and no vacuoles or granules were observed in the cytoplasm. Macrophages were also present in the cavity of the atretic follicle, being larger in size than those in the developing follicle and characterized by cytoplasmic vacuoles and granules of a lysosomal nature. With the luteinization of the follicle, macrophages were seen to be distributed inside and outside the corpus luteum, but constituted only a minor population as compared with other kinds of leukocytes. The intracellular organelles were well-developed, including the lysosomal granules. In early pregnancy, the number of macrophages was noticeably increased in the corpus luteum. They were observed mainly outside the corpus luteum, and stained strongly with hCG immunohistochemically. Macrophages were present in the regressing corpus luteum and in the corpus albicans. Numerous lipid droplets and elongated cholesterol crystals were seen in the cytoplasm. Macrophages therefore appeared to be present throughout the ovarian cycle and may be involved in the development and atresia of the follicles and the progression and the regression of luteal tissues.

Adult↗

Follicular development and ovulation in macrophage colony-stimulating factor-deficient mice homozygous for the osteopetrosis (op) mutation.

To clarify the role of macrophages and macrophage colony-stimulating factor (M-CSF) in follicular development and ovulation, the processes of folliculogenesis and ovulation, numerical changes in macrophages, and proliferative capacity of granulosa cells were examined in op/op mice before or after daily M-CSF administration. The natural estrous cycle was determined daily by means of vaginal smears. The number of ovulated ova in both fallopian tubes was significantly smaller in op/op mice than in normal littermates. Such ova markedly increased in number after daily M-CSF administration. The numbers of both antral and mature follicles in the proestrous ovary were markedly lower in op/op mice than in the controls and increased after daily M-CSF administration. Flash-labeling with [3H]thymidine showed that the proliferative capacity of granulosa cells in antral follicles was reduced in op/op mice but elevated after daily M-CSF administration. Numbers of granulosa cells and macrophages in the antral follicles were significantly decreased in op/op mice but were increased after M-CSF treatment. All these data provide evidence that macrophages are implicated in the process of folliculogenesis and ovulation.

Animals↗

Role of macrophages in ovarian follicular development.

The effects of macrophages on granulosa cell proliferation were examined using gonadotropin-primed immature female rats and osteopetrotic (op/op) mice, a model defective in monocyte-macrophage lineage cells. Macrophages were found in the follicles at various developmental stages in rats and mice. The labeling index with [3H]thymidine of cultured rat granulosa cells was maximal when they were cultured with peritoneal macrophages at a macrophages:granulosa cell ratio of 0.01. This ratio was similar to those in rat preantral and antral follicles in vivo. In op/op mice, the number of developing follicles was markedly reduced, but increased after daily macrophage-colony-stimulating factor (M-CSF) administration. In the antral follicles of op/op mice, both granulosa cells and macrophages were significantly decreased in number but were increased after M-CSF treatment. Double immunohistochemical staining revealed that epidermal growth factor (EGF)-positive cells were macrophages in the developing rat follicles. These findings suggest that macrophages are located in the developing follicles and participate in promoting granulosa cell growth through a paracrine mechanism by secreting EGF and other cytokines.

Animals↗

Mutations in DNA mismatch repair genes are not responsible for microsatellite instability in most sporadic endometrial carcinomas.

Endometrial carcinoma is the second most common tumor type in women with hereditary nonpolyposis colorectal carcinoma. Microsatellite instability (MI) has been observed in the inherited (hereditary nonpolyposis colorectal carcinoma-associated) form of endometrial carcinoma as well as in approximately 20% of presumably sporadic cases. Recent studies suggest that MI in many cell lines or xenografts derived from sporadic colorectal carcinomas is not attributable to mutations in four known human DNA mismatch repair (MMR) genes (hMSH2, hMLH1, hPMS1, and hPMS2). Mutational analyses of these four MMR genes in endometrial carcinomas have not been previously reported. We analyzed nine sporadic MI-positive primary endometrial carcinomas for mutations in the above four MMR genes. Mutations were detected in two tumors (in hMSH2), and both of the mutations were acquired somatically. Immunohistochemical staining revealed a lack of expression of hMSH2 protein in the two tumors containing hMSH2 mutations. Our data suggest that mutations in these four known DNA MMR genes are not responsible for MI in the majority of sporadic endometrial carcinomas displaying this phenotype.

Adult↗

Activated (HLA-DR+) T-lymphocyte subsets in cervical carcinoma and effects of radiotherapy and immunotherapy with sizofiran on cell-mediated immunity and survival.

A prospective randomized trial on 312 patients with locally advanced cervical carcinoma (FIGO stages IB-IV) was carried out. The 5-year survival in 90 patients treated with radiotherapy and antitumor polysaccharide sizofiran, an extract from the culture broth of Schizophyllum commune Fries, in combination was significantly (P = 0.045) better than that in 82 patients treated with radiotherapy alone. Treatment with sizofiran and 5-fluorouracil in combination improved (P = 0.003) the 5-year survival in 60 patients treated with radiotherapy. In 244 cervical carcinoma patients, the percentage of activated CD8+ (CD8+HLA-DR+) T cells in the CD8+ T-cell subsets in peripheral lymphocytes increased significantly as the disease progressed. A similar tendency was observed in the percentage of activated CD4+ (CD4+HLA-DR+) T cells in the CD4+ T-cell subsets. These immunologic parameters were significantly increased by radiotherapy, but not by surgery. Sizofiran accelerated a recovery in the activated CD8+ T cells in the CD8+ T-cell subsets compared with that of sizofiran nontreated patients after radiotherapy. Our data show that possible immune impairment in cervical carcinoma may be caused by disturbances in cell-mediated immunity, and that sizofiran is an effective immunotherapeutic agent for cervical carcinoma because it stimulates a rapid recovery of the immunologic parameters impaired by radiotherapy.

CD4-CD8 Ratio↗

Activated (HLA-DR+) T-lymphocyte subsets in early epithelial ovarian cancer and malignant ovarian germ cell tumors.

We examined peripheral blood T-lymphocyte subsets before initiation of therapy in 79 healthy controls, 3 patients with endometriosis, 95 patients with common epithelial tumors of the ovary, 15 patients with ovarian germ cell tumors, and 3 patients with ovarian sex cord-stromal tumors. In stage Ia/Ib patients with epithelial ovarian cancer, the percentages of activated CD4+ (CD4+HLA-DR+) T cells and activated CD4+ T cells in the CD4+ T-cell subsets were significantly higher than those of healthy controls and patients with benign or borderline epithelial tumors of the ovary. These immunologic parameters were subsequently decreased in patients in stage Ic and more advanced stages. In malignant ovarian germ cell tumors, a similar increase in the CD4+ T-cell subsets was observed. Moreover, the percentage of activated CD8+ T cells in the CD8+ T-cell subsets in stage Ia/Ib patients increased significantly compared with those in healthy controls and patients with benign tumors. Our findings indicate that activated T lymphocytes may play some roles in oncogenesis and progression of both epithelial ovarian cancer and malignant ovarian germ cell tumors.

CD4-Positive T-Lymphocytes↗

Immunohistochemical localization of epidermal growth factor and its effect on granulosa cell proliferation in rat ovary.

The localization of epidermal growth factor (EGF) in the ovary and its effect on proliferation of granulosa cells were investigated in gonadotrophin-primed immature female rats. Immunoreactions with anti-rat EGF monoclonal antibody were observed sparsely in the granulosa layer and antrum of follicles, but not in the theca layer or stromal tissue. The EGF-positive cells were round or oval shaped and often larger than granulosa cells. The localization and morphological appearances of these cells in the follicles were in good agreement with those of macrophages. Although EGF alone did not promote granulosa cell growth in vitro, the labelling index with [3H]thymidine of granulosa cells cultured with 0.1 ng/ml EGF and 0.1 ng/ml basic fibroblast growth factor was significantly greater than that without the growth factors (18.4% vs. 15.8%, P < 0.01). These results suggest that macrophages in follicles may modulate follicular development through a paracrine mechanism by secreting EGF and other growth factors.

Animals↗

Mucinous adenofibroma of the ovary: case report of the endocrinologic findings.

Endocrine and clinicopathologic findings in a 77-year-old woman with ovarian mucinous adenofibroma of borderline malignancy are reported. The preoperative levels of testosterone, androstenedione, estrone, and estradiol in her peripheral blood were 91 ng/ml, 3.78 ng/ml, 82 pg/ml, and 35 pg/ml, respectively, abnormally high. Those of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) were < 0.5 mIU/ml and 12.1 mIU/ml, respectively. Total abdominal hysterectomy and bilateral salpingo-oophorectomy were performed. Pathologic diagnosis of the right ovarian tumor was mucinous adenofibroma of borderline malignancy with hyperthecosis. Postoperatively, the levels of sex steroids and gonadotropin returned to the normal range of a postmenopausal woman. The patient remains well 2 years and 6 months after the operation.

Adenofibroma↗

Isolation, growth and characteristics of human ovarian surface epithelium.

The ovarian surface epithelium (OSE) is a key tissue in the pathogenesis of ovarian surface epithelial-stromal tumours and ovarian endometriosis, commonly encountered gynaecological diseases. Despite the high incidence of these diseases, experimental in vitro studies of OSE are few and so we used the scraping method with an enzymatic procedure to isolate human OSE and studied its characteristics in vitro. Nineteen normal ovaries were used. After incubation of the ovary for 40 min in collagenase type 1 solution (300 U/ml), the surface cells were removed by gentle scraping with a surgical blade. Cells obtained as a cluster after unit gravity sedimentation with 5% bovine serum albumin in medium 199 were cultured in medium 199 containing 15% fetal bovine serum. The viable cell number in a single ovary was 0.1-2.7 x 10(6). The outgrowth of cells started from a homogeneous population of single cells, and the cell population doubling time was between 7 and 10 days. Confluent monolayers were formed after 13-20 days and subcultured from one to three times. The monolayers mostly had a cobblestone appearance, and fusiform or polygonal cells were also observed. By cytochemistry, immunocytochemistry and scanning and transmission electron microscopy, the cells were shown to have characteristics of mesothelial OSE cells in short-term culture. This experimental approach was efficient in providing cultured human OSE, which can be utilized to investigate pathobiology and carcinogenesis.

Adult↗

Scanning electron microscopic and immunohistochemical studies of pelvic endometriosis.

The pathogenesis of pelvic endometriosis has been studied by using scanning electron and light microscopy, observing the surface structure of bluish lesions obtained from 26 patients during laparotomy. Paraffin sections included another 17 tissue samples of endometriosis, based on immunohistochemical responses to epithelial membrane antigen, keratin and vimentin. Ultrastructurally, the surface epithelial cells could not be detected in 13 out of 17 pelvic peritoneal endometriosis samples. In one case in which the surface peritoneal cells were seen histologically to dip into the subperitoneal stroma, many surface peritoneal infoldings were observed, and ciliated cells were detected at the edge of these infoldings. Ovarian endometriosis was composed of three types of cells, none of which had any cilia. These findings were observed in continuity with adjacent normal mesothelial cells. No characteristic structure of the endometrial surface was observed for the bluish lesion, but the gland surface of endometriosis located in the subperitoneal stroma initially had ciliated cells. The immunoreactions in both the columnar mesothelial cells with surface peritoneal infoldings and the glands of endometriotic tissues were similar to those of normal endometrial glands, but different from those of normal mesothelial cells. Pelvic endometriosis might originate by a process of metaplasia from the pelvic peritoneum.

Adult↗