Yoshio Kuwashima

Immunohistochemical characterization of undifferentiated carcinomas of the ovary

Immunohistochemical characteristics of undifferentiated carcinomas of the ovary were examined using formalin-fixed, paraffin-embedded tissues with an avidinbiotin staining approach. Eight cases were collected from the pathology files of our Institute from a total of 214 recorded malignant ovarian tumors. For immunostaining, antibodies reacting with epithelial membrane antigen (EMA), pankeratin, vimentin, CA 125, CA 19-9, carcinoembryonic antigen (CEA), α-fetoprotein (AFP), α-l-antitrypsin (AT), epidermal growth factor receptor (EGFR), c-erbB-2,bcl-2 and p53 proteins were used. All the cases examined were positive for EMA and pankeratin, specific markers for epithelial tumors, negative for the non-epithelial tumor marker, vimentin, and also positive for EGFR. Interestingly, only one case was positive for CA 125, despite it being one of the commonest reported indicators of ovarian cancer. CA 19-9 was positive in 7 cases, CEA in 5, AFP in 2, AT in 6 and c-erbB-2 protein in 4. Two cases were positive for p53 protein, and in 1 of these positive staining forbcl-2 was also observed. These results indicate that the epithelial nature is well preserved in undifferentiated ovarian carcinomas, although consistently positive reactions were not observed within the cases for some antigens. They further celarly show that a negative signal for CA 125 can not be considered to exclude the possibility of a primary ovarian tumor.

Random nuclear p53 overexpression pattern in tamoxifen-mediated endometrial carcinoma.

SummaryIn a previous paper, we suggested that tamoxifen (TAM)-mediated endometrial carcinogenesis may not involve estrogenic pathways because of random estrogen receptor positivity among endometrial carcinomas with and without TAM treatment for breast cancer. DNA adduct formation (reported in rat liver and human endometrium) was considered to be a more plausible mechanism for TAM-mediated carcinogenesis. To examine the reported correlation between DNA adduct formation and p53, the present study examined p53 expression in the endometrial carcinomas reported in the previous study. Seven endometrial adenocarcinomas associated with long-term TAM treatment for breast carcinoma and 4 carcinomas without TAM treatment but with history of breast carcinoma were immunohistochemically investigated for nuclear p53 expression. The bcl-2 product was also examined. Diffuse and intense nuclear reactivity for p53 protein was present in only one TAM-related case. Essentially, no differences were observed in the bcl-2 staining patterns of TAM-treated and -untreated patients with cancer. Thus, p53 overexpression in endometrial carcinomas occurring in patients with breast cancer seems to be not specific for TAM-treated patients, and, if DNA adduct formation has any role in this type of endometrial carcinogenesis, it may not be related preferentially to p53 gene alteration. Further studies are needed to understand the precise mechanism(s) of the endometrial carcinogenesis.

Proliferative and apoptotic status in endometrial adenocarcinoma.

The contribution of cell proliferation and apoptosis to growth patterns in endometrial adenocarcinoma were investigated. Immunohistochemical staining was carried out by an antibody for Ki-67 proliferative antigen, Ley apoptotic antigen, and oncogene products bcl-2 and p53. Forty cases of endometrial adenocarcinoma were classified as exophytic, endophytic, and mixed exo- and endophytic in light of their vertical growth pattern, and, in each case, the carcinomatous area was divided into three layers by its vertical axis. In all but one case, no zonal distribution of the antigen expression was observed. In one case, an exophytic tumor, Ki-67 expression was intense in the surface layer and Ley expression in the deep layer was also intense, suggesting a correlation between macroscopic growth pattern and cellular growth and apoptotic potential. However, in general, zonal distribution of cell proliferation and apoptosis could not explain the growth morphology of endometrial adenocarcinoma of the uterus and it was suggested that factor(s) other than cell proliferation and apoptosis determine macroscopic growth patterns.

Responses of a murine B16 melanoma to pharmacotherapy studied and compared with different assay systems

SummaryThe responses of B16 melanoma in C57BL mice to cytotoxic agents, cyclophosphamide, doxorubicin, and dactinomycin, were studied by different methods both in vivo and in vitro. Either treated tumors were allowed to grow in vivo or cells were isolated and cultured in vitro afterwards. The tumor cells were also treated in vitro and assayed in vitro. For cyclophosphamide, a fairly good correlation was found among the dose responses measured: the tumor growth delay, incidence of giant cells, decrease in mitotic index, clonogenic cell survival of tumor cells treated in vivo and the survival of cells treated in vitro. For doxorubicin and dactinomycin, on the other hand, the tumor growth delay was not marked, the appearance of giant cells and decrease in mitotic index were minimal, and almost no decrease was found in the clonogenic cell survival for tumor treated in vivo, although the cells responded well to in vitro exposure. The results indicate substantial differences in the expression of damage according to the conditions of tumor cell growth and assay after treatment with different agents.

Automated Primary Screening Devices

failure might vary widely. During the past year there has been a lull in the previously very active promotion of primary screening devices. It appears that economic projections for the manufacturers did not materialize. This offers a good opportunity to assess the situation and to suggest where the development might go from here. The first question to be raised requires that we return to the beginning. What is the problem that automation of primary screening is to solve? The often-quoted Wall Street Journal article (Bogdanich W: The pap test misses much cervical cancer through lab’s errors. Nov 2, 1987) called attention to several concerns. There were concerns about the reliability of the reading of a Pap smear, about the possibility of shortages in highly trained personnel and about labor costs. The resulting availability of capital for technology development undoubtedly responded to the need to support the control of cervical cancer, but one has to consider that a prime motivation was the potential for economic gain. There is nothing wrong with that notion in principle, but inevitably it had major consequences. The first consequence was that technical solutions to the problem were explored almost entirely with the medical/economic situation in the United States in mind. A device had to process a slide within a very short time frame so that system throughput would meet revenue projections. Device costs were weighed against amortization schemes, projected unit sales and return on investment capital. There were discussions about acceptable false negative rates and how these might be pegged to revised estimates of false negative rates in current laboratory practice. There were serious and justified concerns about the risk of lawsuits in the United States. One cannot help, though, particularly on the ocAfter 35 years of basic research and development, automated screening devices for cervical cancer went into clinical trials and entered clinical practice. The results have been encouraging and might be called a success in several ways. The United States FDA approved two devices; both proved to perform at a level equal to or better than claimed by the manufacturers. Beyond that the field tests established that it should be possible to design systems that might find approval by the cytopathology community. The devices in the clinical trials were, after all, only first-generation designs. There is no complex technology that performs at the level of its potential until several generations of designs have evolved. As first-generation devices, given the very substantial difficulties of the problem and the circumstances under which their development took place, they performed remarkably well. Eliminating design compromises dictated either by the state of technology at the time of development or by the development climate concomitant with venture capital financing would produce the next generation of primary screening systems that come much closer to a performance level well respected by medical professionals. One might hear the argument that since performance of the first-generation devices was critically assessed by the FDA in the United States, why should the profession call for improvements? It is true that the FDA approved these devices. However, one has to understand that the FDA here merely certified that the devices perform at the level claimed by the manufacturer. If one wanted to be unkind, one might suggest that the industry set its own standard. In practice it would essentially be the laboratory director who decides whether a claimed performance level can be maintained and is acceptable or not. The extent to which such a decision would be based on a thorough understanding

Rhabdomyosarcoma with FocaI Cartilaginous Differentiation (Malignant Mesenchymoma) of the Inferior Vena Cava

A sarcoma arising from the inferior vena cava occupied the entire lumen of the inferior vena cava, right atrium, hepatic veins and conmon iliac veins. Its histological appearance was non‐ specific sarcoma, except for the presence of a few rhabdomyoblasts and some immature cartilaginous tissue. lmmunohistochemically, some tumor cells were positive for myoglobin, desmin, HHF‐35, and vimentin. Electron microscopy revealed that some tumor cells contained myofilaments and Z bands in the cytoplasm, which are characteristics of rhabdomyosarcoma. The tumor was diagnosed as rhabdomyosarcoma with focal cartilaginous differentiation (malignant mesenchymoma) of the inferior vena cava. Acta Pathol Jpn 42: 382–385, 1992.

Neoplastic Argentaffin Cells with Intracytoplasmic Eosinophilic Granules in a Gastric Adenocarcinoma

A case of poorly differentiated adenocarcinoma of the stomach with unique histological features is reported: in addition to characteristic adenocarcinoma cells, a large number of tumor cells contained bright eosinophilic and argentaffin granules in their cytoplasm. On routine histologic examination, the latter cells closely resembled the endocrine cells present in the normal human gastrointestinal tract, although the granules were distributed throughout the cytoplasm and did not show any polarity, which is usually subnuclear in normal endocrine cells. Imuunohis‐tochemical studies demonstrated positive staining for lysozyme, CEA, gastrin and HCG. Electron microscopic examination revealed cytoplasmic neurosecretory granules, and some tumor cells were found to contain both secretory granules and mucinous material within the same cytoplasm. These neoplastic endocrine cells presumably originated from primitive digestive system elements capable of differentiating towards both endocrine and mucus‐secreting varieties. Acta Pathol Jpn 41: 905‐910, 1991.

In vivo dynamics of pulmonary lymphoid cell subpopulations generated against pulmonary metastasis : evaluation by bronchoalveolar lavage fluid

SummaryThe dynamics of lymphoid cell subpopulations in bronchoalveolar lavage fluid (BALF) and the systemic lymphoid organs of mice after intravenous injection of B16 melanoma cells were examined with a fluorescence-activated cell sorter. The lymphoid cell subpopulations of BALF and mediastinal lymph nodes showed significant changes in numbers and proportions, while those of other lymphoid organs including inguinal lymph nodes, thymus and spleen, showed little change. In week 1, the cells with a Thy-1.2+, Lyt-1+, L3T4−, Lyt-2− phenotype and asialo-Gm1+ cells in BALF significantly increased and L3T4+ cells slightly increased in number. By week 3, the numbers of Lyt-2+ cells in BALF markedly increased in number (by about 90 times) compared with controls. The number of Thy1.2+ cells in mediastinal lymph nodes also increased significantly by week 3. Mice that had been pretreated with an immunosuppressive dose of cyclophosphamide were also inoculated intravenously with B16 melanoma cells. In these mice, a significantly increased number of pleural tumors developed and the number of Thy-1.2+ cells in BALF was markedly reduced from week 1 to 3. The results indicate that L3T4 and Lyt-2 double negative T-cells and natural killer (NK) cells may be generated and/or mobilized to the lung in an early phase of experimental metastasis of B16 melanoma cells and that at a later stage, when multiple metastases develop, T-cells with a Lyt-2+ phenotype markedly increase, probably as an expression of a host reaction against proliferating metastatic tumor cells.

Fine needle aspiration cytology of adenocarcinoma of the rectovaginal septum : A case report

BACKGROUND Adenocarcinoma arising in the rectovaginal septum is exceedingly rare and is difficult to diagnose by pathologic examination prior to surgery because of the anatomic position of the tumor. CASE A 42-year-old woman presumed to have adenocarcinoma of the rectovaginal septum underwent fine needle aspiration for diagnosis. Although a previously performed biopsy from the posterior vaginal fornix was unsuccessful, fine needle aspiration cytology via the posterior vaginal wall detected adenocarcinoma cells. The cell clusters were composed of cells with enlarged and hyperchromatic nuclei. The nuclei themselves demonstrated round and/or irregular morphologic patterns, with high nuclear/cytoplasmic ratios, and often contained an enlarged, round nucleolus and sometimes multiple ones in a single nucleus. Aniso-nucleosis was severe, and the chromatin patterns ranged from coarse to finely granular. The cytoplasm was narrow and lightly stained. Following fine needle aspiration, the patient underwent posterior exenteration on the basis of the cytologic diagnosis. CONCLUSION Fine needle aspiration cytology was useful in establishing the preoperative diagnosis of adenocarcinoma of the rectovaginal septum, and curative exenterative surgery could be then performed. To our knowledge, this is the first report of fine needle aspiration cytology of adenocarcinoma at this location.

Fine needle aspiration cytology of argyrophilic mucinous carcinoma of the male breast

The results of fine needle aspiration (FNA) cytology of a case of argyrophilic mucinous carcinoma of the male breast is presented. The smear obtained from the tumor showed many loose clusters of atypical cells with abundant granular cytoplasm in the mucous background. Most tumor cells showed a plasmacytoid appearance with eccentric nuclei. Numerous fine granules and a few large droplets were found in the cytoplasm. The characteristic light microscopic feature of this lesion was organoid nest formation of tumor cells in mucous lakes. The Grimelius method revealed numerous argyrophilic granules in the cytoplasm and immunohistochemistry showed a positive reaction for chromogranin A. Thus, this tumor revealed the cytomor-phologic characteristics of mucinous as well as argyrophilic carcinoma, simultaneously.