Masanori Yasuda

Expression and Localization of Chromogranin A Gene and Protein in Human Submandibular Gland

Human saliva chromogranin A (CgA) is clinically promising as a psychological stress marker. However, expression of CgA is poorly understood in humans, although salivary gland localization of CgA in other mammals, such as rodents and horses, has been demonstrated. In the present study, we investigated the expression and localization of CgA in the human submandibular gland (HSG) using various methods. CgA was consistently localized in serous and ductal cells in HSG, as detected by immunohistochemistry and in situhybridization. Reactivity was stronger in serous cells than in ductal cells. In addition, strong immunoreactivity for CgA was observed in the saliva matrix of ductal cavities. Western blotting gave one significant immunoreactive band of 68 kDa in the adrenal gland, HSG and saliva. Finally, CgA was detected in secretory granules of serous and ductal cells by immunoelectron microscopy. In conclusion, CgA in humans is produced by HSG and secreted into saliva.

Neuroendocrine marker expression in thyroid epithelial tumors

Tissue sections from 50 cases with thyroid tumors, composed of 11 follicular adenomas, 10 follicular carcinomas, 14 papillary carcinomas, 10 anaplastic carcinomas, and 5 medullary carcinomas, were immunohistochemically analyzed for representative neuroendocrine markers. Immunoexpression ratios of these neuroendocrine markers were as follows: Follicular adenomas, neuron-specific enolase (NSE) 63.6%, synaptophysin (SynP) 45.5%, Leu7 27.3%, NCAM 45.5%, chromogranin A (CgA) 0%, SNAP25 0%; follicular carcinomas, NSE 90.0%, SynP 80.0%, Leu7 80.0%, NCAM 0%, CgA 0%, SNAP25 0%; papillary carcinomas, NSE 85.7%, SynP 78.6%, Leu7 100%, NCAM 7.0%, CgA 0%, SNAP25.0%; anaplastic carcinomas, NSE 10.0%, SynP 0%, Leu7 0%, NCAM 0%, CgA 0%, SNAP25 0%; medullary carcinomas, NSE 100%, SynP100%, Leu7 80.0%, NCAM 40.0%, CgA 100%, SNAP25 100%. The two follicular carcinomas, which were morphologically characterized by insular (or alveolar) arrangements, showed distinct immunoexpression of NSE and SynP at the same time. By in situ hybridization (ISH), expression of mRNA for NSE was confirmed in cases with marked immunoexpression of NSE. Although no endocrine granules were found, our results suggested that a specific type of follicular carcinoma, i.e., insular variant, may be immaturely neuroendocrine-differentiated.

Excessive formation of basement membrane substance in clear‐cell carcinoma of the ovary: Diagnostic value of the “raspberry body” in ascites cytology

Formation of basement membrane‐like substance or so‐called collagen core was characteristic of clear‐cell carcinoma of the ovary in ascites cytology. The hyaline extracellular material was stained light green in Papanicolaou smears and pinkish to purplish red in Giemsa preparations and was frequently observed within the cancer cell clusters in all ten samples of clear‐cell carcinoma. Such a structure termed “raspberry body” was focally seen in one of 30 specimens of serous cystadenocarcinoma and one of 30 samples containing reactive mesothelial cells. The “raspberry body” was not found in ascitic fluid from ten patients with mucinous cystadenocarcinoma and two with endometrioid carcinoma. Overproduction of the acidic‐charged basement membrane substance was confirmed by 1) cytochemical positivity for periodic acid‐Schiff, alcian blue (pH 2.5 or pH 1.0), colloidal iron, and periodic acid‐methenamine silver, 2) immunocytochemical demonstration of laminin and type 4 collagen, and 3) ultrastructural identification of excessive formation of the basal lamina. Recognition of the “raspberry body” helps cytopathologists make a cytologic diagnosis of this chemotherapy‐resistant malignancy disseminated in ascitic fluid. Diagn. Cytopathol. 16:500–504, 1997.

Immunohistochemical Study of Type-1 Blood Antigen Expressions in Thyroid Tumors: The Significance for Papillary Carcinomas

Immunohistochemical expressions of type 1 blood group antigens were studied for 95 cases of thyroid tumors, including 29 follicular adenomas, 23 follicular carcinomas, and 43 papillary carcinomas, applying monoclonal antibodies against DU-PAN-2, CA19–9, Lewisa (Lea), and Lewisb (Leb). Normal thyroid tissue invariably failed to express all four antigens. In follicular adenomas, DU-PAN-2 and CA19–9 were focally expressed in 7% and 21% of cases, and in follicular carcinomas, CA19–9 expression was limited to one case (4%); all cases were negative for DU-PAN-2. No or little expression of Lea or Leb was observed in these follicular tumors. In contrast, DU-PAN-2, CA19–9, Lea, and Leb were expressed in 98%, 84%, 33%, and 49% of 43 papillary carcinomas, respectively. The positive stainings were observed mainly on the luminal surface of the tumor cells. The number of tumor cells that expressed DU-PAN-2 generally was greater than that of tumor cells that expressed CA19–9, Lea, or Leb. There was no significant difference in antigen expressions in female papillary carcinomas between subjects who were younger and older than 50 years old. The results suggest that DU-PAN-2 would be a useful immunohistochemical marker for distinguishing papillary carcinomas from follicular tumors. These immunohistochemical profiles imply the following: the activity of α2–3 sialyltransferase, a specific glycosyltransferase, would be more strongly enhanced in papillary carcinomas than in follicular tumors; the antigen expressions in papillary carcinomas may not be related to the alteration of the female sex hormone environment.

Cytologic Study of Ascites and the Endometrium in Ovarian Carcinoma

OBJECTIVEnTo ascertain the usefulness of endometrial cytology with ovarian cancers when examining extension of the disease and to analyze significant factors associated with migration of ovarian cancer cells into the uterine cavity.nnnSTUDY DESIGNnCytologic results on ascites and the endometrium were analyzed in 87 patients with primary ovarian cancer in the absence of metastasis to the endometrium or cervicovagina.nnnRESULTSnPositive results for cytology were found in 62/87 of ascites cases (71.3%) and in 20/87 endometrium cases (23.0%). The 15 cases (15/62 or 24.2%) positive for ascitic and endometrial cytology, divided clinically into stage III (6 cases) and stage IV (9 cases), were classified histologically as serous, 7 cases; mucinous, 2 cases; clear cell, 4 cases; endometrioid, 1 case; and unclassified, 1 case. Half the clear cell carcinomas (4/8 or 50.0%) were positive in the ascites and endometrium. The ascitic volume at surgery exceeded 500 mL in 9/15 cases (60.0%).nnnCONCLUSIONnPapillae with basement membrane material in the cores may be structurally associated with migration of ovarian cancer cells into the uterine cavity, especially in clear cell carcinomas. Cytologic positivity of the endometrium and ascites significantly correlated with ascitic volume.

Subcellular organelle analysis applying confocal laser scanning microscopy to the steroid‐producing epithelial ovarian tumor: A technical note

An epithelial ovarian tumor with steroid production was examined in a 70‐year‐old postmenopausal female. The stromal cells of this tumor were rather dense and occasionally characterized by tuteinization or hyperthecosis, which has been associated with steroidogenesis. Subcellular visualization using confocal laser scanning microscopy (CLSM) successfully led to the identification of 3β‐hydroxysteroid dehydrogenase (HSD) in both mitochondria‐like small particles and endoplasmic reticulum‐like linear profiles, and P450‐aromatase also in endoplasmic reticulum‐like linear profiles, on the three‐dimensional images.

Treatment Strategy for Recurrent and Refractory Epithelial Ovarian Cancer: Efficacy of High-Dose Chemotherapy with Hematopoietic Stem Cell Transplantation

According to population statistics in Japan, approximately 3,800 women die of ovarian cancer annually, and approximately 6,000 are affected by this disease. Ovarian cancer is referred to as a “silent tumor”, since patients have few subjective symptoms and by the time symptoms are observed, the cancer has progressed to Stage III or IV in about half of the patients. The basic treatment for advanced epithelial ovarian cancer is to remove as much of the tumor as possible, and subsequently to perform anticancer therapy using drugs such as cisplatin, carboplatin and paclitaxel, all of which have been shown to be effective for epithelial ovarian cancer. However, the 5-year survival rate in advanced ovarian cancer patients is still only about 20%, and a treatment that leads to long-term survival has yet to be developed. Here, we review the available treatments for ovarian cancer, and present the results of high-dose chemotherapy (HDC) performed in our hospital for recurrent and refractory ovarian cancer.

Long-term results and prognostic analysis in advanced and recurrent/refractory epithelial ovarian cancer treated by high-dose cyclophosphamide, adriamycin, and cisplatin with autologous bone marrow transplantation

AbstractBackground. The efficacy of high-dose chemotherapy (HDC) with autologous bone marrow transplantation (ABMT) was evaluated in patients with advanced or recurrent/refractory (r/r) epithelial ovarian cancer in terms of long-term results and prognostic analysis.nMethods. Between 1984 and 1991, 47 patients were prescribed two courses of HDC, consisting of cyclophosphamide (1600–2400 mg/m2), adriamycin (80–100 mg/m2), and cisplatin (100–150 mg/m2) after maximal cytoreductive surgery. Prior to HDC, platinum-based chemotherapy was administered for optimal cytoreduction.nResults. The 5- and 8-year overall survival (OS) rates (%) of the 47 patients were 44.7% and 40.4%, and the 5- and 8-year disease-free survival (DFS) rates (%) were 29.8% and 27.7%, respectively. The 5- and 8-year OS rates (%) by stage were: stage III, 60.0% and 52.0%; stage IV, 33.3% and 33.3%; and r/r, 20.0% and 20.0%. The 5- and 8-year DFS rates (%) by stage were: stage III, 40.0% and 36.0%; stage IV, 25.0% and 25.0%; and r/r, 10.0% and 10.0%, respectively. Significantly better long-term survival (P < 0.01) was obtained in the group with no residual disease or residual disease <0.5 cm than in the groups with residual disease of 0.5–2 cm and >2 cm. In the 32 stage III/IV patients, the group given two courses of 150 mg/m2 cisplatin (n = 18) showed significantly better long-term survival (P < 0.05) than another group given two courses of 100 or 120 mg/m2 cisplatin (n = 14).nConclusions. Administration of two courses of HDC con-taining 2400 mg/m2 cyclophosphamide, 100 mg/m2 adriamycin, and 150 mg/m2 cisplatin per course followed by ABMT appears to be a promising procedure for achieving long-term survival in patients with chemosensitive advanced or r/r epithelial ovarian cancers with no or minimal residual disease.

A kidney carcinoma with features of clear cell renal carcinoma and transitional cell carcinoma: a combined renal cell and transitional cell carcinoma?

Renal cell carcinoma (RCC) arises from kidney parenchyma, while transitional cell carcinoma (TCC) from the renal pelvis in the kidney. We report a kidney carcinoma with histological features of RCC and TCC. A 72-year-old Japanese man presented with right lumbar pain and hematuria, and was admitted to our clinic. Imaging modalities showed a solid tumor in the upper half of the right kidney. RCC was suspected, and total nephrectomy was performed. Grossly, the tumor was solid and yellowish tan with a rather hard consistency. The tumor measured 5 6 5 cm and invaded into the pericapsular fat tissue and renal pelvis. Histologically, there were two populations of tumor cells. One component was composed of clear cells with hyperchromatic nuclei arranged in a trabecular pattern (Fig. 1A). This was interpreted as RCC of clear cell type. The other component consisted of stratified cells with hyperchromatic nuclei and dark cytoplasm (Fig. 1B). This was diagnosed as TCC. These two components were admixed in the tumor (Fig. 2A), and gradual transition between the two was recognized in places (Fig. 2B). The distribution of both components was even in the tumor. The renal pelvis and extracapsular invading front showed the same morphology. The RCC component comprised approximately 50% of the tumor, and the TCC component another 50%. Vascular and lymphatic invasion was recognized in some areas. The non-tumorous kidney showed no significant changes. The tumor cells of both components were negative with periodic acid–Schiff base (PAS), alcian blue, and Grimelius stains. Immunohistochemically, the tumor cells of both components showed the same results. They were positive for low-molecular-weight cytokeratins (LMWCK) using antibodies (KL-1, MNF116, and CAM5.2) and epithelial membrane antigen, but were negative for high-molecular-weight CK (HMWCK) (AE1/AE3), vimentin, carcinogenic embryonic antigen (CEA), desmin, alpha-smooth muscle antigen, S-100 protein, chromogranin A, CD68, C-erbB2, p53, bcl-2, and placental alkaline phosphatase. CK profiling showed that both components of the tumor cells were positive for CK7, CK18 and CK19, while they were negative for CK8 and CK20. Ki-67 labeling was approximately 15%. RCC and TCC are two major carcinomas occurring in the kidney. RCC may arise from proximal convoluted tubules, thus developing in the parenchyma of the kidney [5]. In contrast, TCC arises from urothelia of the renal pelvis and calices, thus occurring in the renal pelvis. Embryologically, both renal parenchyma and pelvis develop from mesodermal metanephros [5]. Renal parenchyma develops from metanephric tissue (nephrogenic blastema), and the renal pelvis from the ureteric bud. Thus, renal parenchyma and pelvis have common developmental origin. Histologically, the present tumor was composed of RCC and TCC. Both elements were admixed in the tumor, and there were gradual transitions between the two. Therefore, we diagnosed the tumor as combined RCC and TCC. A search of the English literature failed to detect such a combined RCC and TCC, though there were several cases of RCC and TCC that were separately detected in the same kidney [2, 4]. Such an entity as the T. Terada ()) Department of Pathology, Shizuoka Municipal Shimizu Hospital, Shimizu-Miyakami 1231, 424-8636 Shizuoka, Japan e-mail: byouri@city.shimizu.shizuoka.jp Fax: +81-543-361315

Double squamous cell carcinomas, verrucous type and poorly differentiated type, of the urinary bladder unassociated with bilharzial infection

A case of a 66‐year‐old Japanese woman with two synchronous urinary biadder tumors, namely verrucous carcinoma and poorly differentiated squmamous cell carcinoma, is described. Both tumors were accompanled by widespread squamous metaplasia in the background and unassociated with bilharzial infection. The poorly differentiated squamous cell carcinoma, having a satellite tumor in Its proximlty, was large and in an advanced stage. The verrucous carcinoma was small with minimal invasion to the muscuiaris propria. The boundary between both tumors was well defined, suggesting colliding growth appearance. Immunoexpression of cytokeratins of verrucous carcinoma was simllar to that of squamous metaplasia, and significant differences between verrucous carcinoma and poorly differentiated squamous cell carcinoma were demonstrated in thair immunoexpression of cytokeratins.