Ultrastructural and Immunohistochemical Contribution to the Histogenesis of Human Cardiac Myxoma

The ultrastructural features of 8 human cardiac myxomas were analyzed and correlated with immunohistochemical data, with the aim to clarify the characteristics of the cell lines involved in the tumor genesis. Immunohistochemical studies were performed to detect the presence and the distribution of intracytoplasmic filaments (vimentin, desmin, actin, myosin) as well as myoglobin and factor VIII-related antigen, albumin, and lysozime. Eighty percent of myxoma cells were simultaneously positive for vimentin, desmin, and actin, whereas 30% of them stained with antifactor VIII and antivimentin antibodies. The submicroscopic analysis revealed two main cell populations: (1) one composed of stellate-shaped cells with scanty organelles and sparse hyaloplasmic filaments scattered throughout the myxoid stroma and forming a loose network with their projections; (2) another one included cells with more cytoplasmic organelles, intermediate filaments, and myofilaments arranged either singly or in both solid and hollow cord-like structures. Our results support the hypothesis that cardiac myxoma may originate from a reserve multipotent mesenchymal cell able to differentiate more or less completely along two major evolutional lines: myoid and endothelial. The tumor tissue thus seems to be involved in vessel formation, suggesting a growth pattern akin to that manifested in other forms of endocardial pathological reactivity in which reserve mesenchymal cells are engaged.     

Ultrastructural and Immunohistochemical Contribution to the Histogenesis of Human Cardiac Myxoma

The ultrastructural features of 8 human cardiac myxomas were analyzed and correlated with immunohistochemical data, with the aim to clarify the characteristics of the cell lines involved in the tumor genesis. Immunohistochemical studies were performed to detect the presence and the distribution of intracytoplasmic filaments (vimentin, desmin, actin, myosin) as well as myoglobin and factor VIII-related antigen, albumin, and lysozime. Eighty percent of myxoma cells were simultaneously positive for vimentin, desmin, and actin, whereas 30% of them stained with antifactor VIII and antivimentin antibodies. The submicroscopic analysis revealed two main cell populations: (1) one composed of stellate-shaped cells with scanty organelles and sparse hyaloplasmic filaments scattered throughout the myxoid stroma and forming a loose network with their projections; (2) another one included cells with more cytoplasmic organelles, intermediate filaments, and myofilaments arranged either singly or in both solid and hollow cord-like structures. Our results support the hypothesis that cardiac myxoma may originate from a reserve multipotent mesenchymal cell able to differentiate more or less completely along two major evolutional lines: myoid and endothelial. The tumor tissue thus seems to be involved in vessel formation, suggesting a growth pattern akin to that manifested in other forms of endocardial pathological reactivity in which reserve mesenchymal cells are engaged.     

Ovarian Myxoma: Ultrastructural and Immunohistochemical Findings

The ultrastructural and immunohistochemical findings are reported in two ovarian myxomas, one of which was also associated with a sclerosing stromal tumor of the same ovary. Both neoplasms showed a myxoid, moderately cellular proliferation of spindle and stellate cells interspersed with areas of fibrosis and hemorrhage as well as delicate vascular spaces. Ultrastructurally, stellate neoplastic cells with irregular nuclei and occasional nucleoli were embedded in a mucinous and loose collagen matrix. Their cytoplasm showed abundant intracytoplasmic thin filaments that rarely condensed into poorly formed dense bodies. These thin filaments correlated with immunoreactivity for musclespecific actin and vimentin. The neoplastic cells showed no immunoreactivity with antibodies to desmin, S-100 protein, cytokeratin AE1 :AE3, factor Vlll-related protein, or placental alkaline phosphatase. These ultrastructural and immunohistochemical findings are consistent with myofibroblastic differentiation. These ancillary studies exclude important, clinically more aggressive differential considerations such as myxoid rhabdomyosarcoma, myxoid liposarcoma, myxoid neural tumors, mucinous adenocarcinoma showing pseudomyxomatous change, and myxoid-appearing endodermal sinus (yolk sac) tumor.     

Warthin's Tumor: An Ultrastructural and Immunohistochemical Study of Basilar Epithelium

The cellular characteristics of the basilar epithelium in Warthin's tumor have had limited investigation. Ultrastructural examination of basal cells in 9 Warthin's tumors reveals that in addition to numerous mitochondria these cells possess a rich complement of tonofilaments. However, in three examples there are a proportion of these tonofilament-rich cells that have a narrow band of microfilaments in the peripheral cytoplasm adjacent to the basal lamina. Frozen sections of Warthin's tumor and normal salivary glands, doubly labeled with rhodamine-phalloidin for actin and monoclonal antibody 312C8-1 for cytokeratin 14, show that normal myoepithelial cells of acini and intercalated ducts have both of these filaments, as do a proportion of basal cells in the tumor. There are distinct differences in the cytokeratin polypeptide complement between normal luminal and myoepithelial cells as well as between luminal and basal cells in Warthin's tumor. Differences occur in the cytokeratin profiles between the luminal and basal cells of Warthin's tumor and comparable cells in the normal gland; however, there continue to be some similarities in the cytokeratin polypeptides of myoepithelium and the basal cells of normal salivary ducts and the basal cells of Warthin's tumor. These findings show that basal cells in Warthin's tumor are a mixed population with some capable of differentiating as myoepithelial-like cells, and that this tumor could arise from any level of the normal salivary gland duct system.     

Warthin's Tumor: An Ultrastructural and Immunohistochemical Study of Basilar Epithelium

The cellular characteristics of the basilar epithelium in Warthin's tumor have had limited investigation. Ultrastructural examination of basal cells in 9 Warthin's tumors reveals that in addition to numerous mitochondria these cells possess a rich complement of tonofilaments. However, in three examples there are a proportion of these tonofilament-rich cells that have a narrow band of microfilaments in the peripheral cytoplasm adjacent to the basal lamina. Frozen sections of Warthin's tumor and normal salivary glands, doubly labeled with rhodamine-phalloidin for actin and monoclonal antibody 312C8-1 for cytokeratin 14, show that normal myoepithelial cells of acini and intercalated ducts have both of these filaments, as do a proportion of basal cells in the tumor. There are distinct differences in the cytokeratin polypeptide complement between normal luminal and myoepithelial cells as well as between luminal and basal cells in Warthin's tumor. Differences occur in the cytokeratin profiles between the luminal and basal cells of Warthin's tumor and comparable cells in the normal gland; however, there continue to be some similarities in the cytokeratin polypeptides of myoepithelium and the basal cells of normal salivary ducts and the basal cells of Warthin's tumor. These findings show that basal cells in Warthin's tumor are a mixed population with some capable of differentiating as myoepithelial-like cells, and that this tumor could arise from any level of the normal salivary gland duct system.     

Cardiac Myxoma: Histochemical and Ultrastructural Localization of Glycosaminoglycans and Proteoglycans

A recurrent cardiac myxoma is examined histochemical ly at the ultrastructural level. By routine electron microscopy the stellate “myxoma” cell exhibits features suggestive of a secretory function in synthesis of its myxoid stroma. Spicer's high iron diamine (HID), which stains specifically for sulfated glycoconjugates, is utilized for intracellular localization of glycosaminoglycans. HID-positive reactive sites are localized within the Golgi-derived vacuoles and secretory granules of the myxoma cells. No staining is obtained with other cytoplasmic organelles except rare secondary lyso-somes. Although colloidal iron is less specific, both intracellular and extracellular positive reactive sites are observed. With ruthenium red staining the proteoglycans in the extracellular stroma can be visualized as numerous positively stained, polygonal 250-500 A matrix granules with faint filamentous projections. Positive intracellular ruthenium red-stained granules are also observed within the Golgi-derived vacuoles. The alcianophilia of the myxoid stroma with Alcian blue is almost completely abolished by prior treatment with bovine testicular hyaluronidase but is unaffected by leech hyaluronidase, indicating chondroitin sulfates A and/or C, not hyaluronic acid, as the major biochemical constituents of the stroma and the observed extracellular matrix granules. The above findings provide cytochemical evidence of intracellular synthesis of sulfated glycosaminoglycans and proteoglycans of the myxoma cell and its active participation in production of its stroma.     

Cardiac Myxoma: Histochemical and Ultrastructural Localization of Glycosaminoglycans and Proteoglycans

A recurrent cardiac myxoma is examined histochemical ly at the ultrastructural level. By routine electron microscopy the stellate “myxoma” cell exhibits features suggestive of a secretory function in synthesis of its myxoid stroma. Spicer's high iron diamine (HID), which stains specifically for sulfated glycoconjugates, is utilized for intracellular localization of glycosaminoglycans. HID-positive reactive sites are localized within the Golgi-derived vacuoles and secretory granules of the myxoma cells. No staining is obtained with other cytoplasmic organelles except rare secondary lyso-somes. Although colloidal iron is less specific, both intracellular and extracellular positive reactive sites are observed. With ruthenium red staining the proteoglycans in the extracellular stroma can be visualized as numerous positively stained, polygonal 250-500 A matrix granules with faint filamentous projections. Positive intracellular ruthenium red-stained granules are also observed within the Golgi-derived vacuoles. The alcianophilia of the myxoid stroma with Alcian blue is almost completely abolished by prior treatment with bovine testicular hyaluronidase but is unaffected by leech hyaluronidase, indicating chondroitin sulfates A and/or C, not hyaluronic acid, as the major biochemical constituents of the stroma and the observed extracellular matrix granules. The above findings provide cytochemical evidence of intracellular synthesis of sulfated glycosaminoglycans and proteoglycans of the myxoma cell and its active participation in production of its stroma.     

Placental Site Trophoblastic Tumor of the Uterus: An Ultrastructural and Immunohistochemical Study and Immunohistochemical Study

Ultrastructural and immunohistochemical studies in a case of placental site trophoblastic tumor (PSTT) of the uterus were carried out in order to define the nature of the abnormal tissue. By electron microscopy, the large cells, whether mononuclear or syncytial, showed numerous ribosomes, prominent Golgi elements, and abundant rough endoplasmic reticulum (RER) filled with granular material. Pseudopods and microvilli were found on the cell surfaces. By immunofluorescence, the well-developed filamentous cyto-skeleton proved to be actin-rich. (3-HCG (human chorionic gonadotropin) and SP, (0,-specific pregnancy glycoprotein) were detected in only a few tumor cells, whereas most of them stained for HPL (human placental lactogen). The present results show the secretory nature of most of the tumor cells, which resemble the intermediate trophoblast of the placental bed. Together with previous studies, they suggest that a varying spectrum of syncytiotrophoblastic differentiation exists in PSTT. Decidual, myometrial, or histiocytic cells do not seem involved in the histogenesis of the tumor tissue.     

“Aberrant Elastic” in Elastofibroma: An Immunohistochemical and Ultrastructural Study

Elastofibroma is a rare lesion characterized by the presence of abundant abnormal elastic fibers with a unique morphology, fibroblastic proliferation, and collagen deposition. Whether the altered morphology of the elastic fibers is a degenerative phenomenon or is due to abnormal elastogenesis is controversial. We studied fetal skin and three cases of elastofibroma by light microscopy and immunohistochemistry using an antibody to lysozyme, and one case of elastofibroma by electron microscopy (EM). Our previous studies have shown that normal elastic fibers in adult skin do not stain for lysozyme whereas abnormal elastic fibers in solar elastosis and pseudoxanthoma elasticum react positively for lysozyme. In the fetal skin and all three cases of elastofibroma the elastic fibers were negative for lysozyme. EM showed the abnormal flowerlike configuration of the elastic fibers, which consisted of a central core of normal or degenerating elastin surrounded by radiating spokes of granular and filamentous material of variable electron densities, suggesting that the structure and organization of the microfibrils is abnormal. The absence of lysozyme in the aberrant elastic did not differentiate whether there was excessive production of fetal or adult elastic. However, the excessive amount of microfibrils seen at the ultrastructural level suggests that there may be excessive fetal elastic production. The elastic fibers were intimately related to the fibroblasts and were often present within their caveolae, suggesting that the abnormal elastic fibers are produced by the fibroblast. Our study suggests that abnormal elastogenesis with subsequent degeneration plays a role in the production of the abnormal elastic fibers in elastofibroma.     

Mucin-Positive Epithelial Mesotheliomas: A Histochemical, Immunohistochemical, and Ultrastructural Comparison with Mucin-Producing Pulmonary Adenocarcinomas

Pathologists routinely use histochemistry, immunohistochemistry, and electron microscopy to differentiate epithelial mesotheliomas from pulmonary adenocarcinomas. Epithelial mesotheliomas are usually mucicarmine-, PAS-diastase, and carcinoembryonic antigen-negative, whereas about 60-75% of pulmonary adenocarcinomas are mucicarmine- and PAS-diastase-positive, and about 90% express polyclonal carcinoembryonic antigen. During a pathologic evaluation of pleural neoplasms between 1975 and 1990, 10 epithelial mesotheliomas were identified that were mucicarmine- and in some instances PAS-diastase-positive (diagnosis of mesothelioma confirmed by ultrastructural examination), with four mesotheliomas focally expressing carcinoembryonic antigen. The mucicarmine, PAS-diastase, and carcinoembryonic antigen staining were usually eradicated or reduced in intensity by pretreatment of the tissue sections with hyaluronidase, suggesting that hyaluronic acid was responsible for the positive mucin reactions. In three cases the epithelial mesotheliomas showed focal regions of mucicarmine, PAS-d-, and Alcian blue-hyaluronidase-resistant staining. In contrast, 10 mucicarmine-, PAS-diastase-, Alcian blue-, and carcinoembryonic antigen-positive pulmonary adenocarcinomas were not affected by hyaluronidase pretreatment of the tissue. Besides the usual ultrastructural features of well- to moderately well-differentiated epithelial mesotheliomas, the mucin-positive epithelial mesotheliomas often showed medium-electron-dense secretory material covering the microvilli, aggregates of medium electron-dense material in association with the microvilli, producing an ultrastructural morphology that has been observed only in epithelial mesotheliomas.     

Intriguing Case: Pulmonary Blastoma: An Ultrastructural and Immunohistochemical Study with Special Reference to Nuclear Filament Aggregation

A case is presented of pulmonary blastoma occurring in the right upper lobe of a 25-year-old man without distinct clinical features and laboratory abnormality. Light microscopic analysis revealed that the tumor was composed of branching glands and morulae embedded in a primitive but bland mesenchyme. Immunohistochemically the epithelial cells were im-munoreactive for cytokeratins, S-100 protein, protein gene product 9.5, chromogranin A, calcitonin, and Ki-67 (MIB-1); the mesenchymal cells were immunoreactive for vimentin, actin, cytokeratins, and Ki-67; and all the tumor cells were negative for p53, estrogen receptor protein, and human chorionic gonadotropin ß. Characteristically, many epithelial cells contained optically clear nuclei which were immunoreactive for biotin (M743). Electron microscopic analysis revealed that the optically clearing change was due to replacement of the central area of the nuclei by a mass of parallel-arranged 7- to 10-nm filaments, and biotin-immunoreactive products were mainly localized in the nuclear matrix. Additionally, spherical bodies were identified in the cytoplasm of the nuclear filament-aggregated cells, suggestive of an intimate pathogenetic association of the two morphological abnormalities. The similarity of the aggregated nuclear filaments to those observed in gestational endometrium and ovarian endometrioid carcinoma implies that a similar mechanism plays a role in the pathogenesis of these abnormalities.     

Ovarian Myxoma: Ultrastructural and Immunohistochemical Findings

The integrity of the colonic mucin layer has been reported to be altered during carcinogenesis in both humans and rodents. Prior to attempting scanning microscopic techniques on colonic mucosa of patients at high risk to develop colorectal cancer, these procedures were performed on colonic mucosa from rats with chemically induced colon cancers. Substantial technical difficulties in preparation and serious subjectivity in interpretation of the scanning micrographs were encountered. The major technical problem was the unpredictable retention of the mucin layer upon both normal and cancerous mucosae. Visual interpretation of the integrity or disruption of the mucin layer with the scanning electron microscope revealed variable fenestration and fraying of the mucin in both normal and cancerous colons. Our findings suggest that scanning electron microscopy of colonic mucin may not be a reliable screening procedure for (pre)cancerous changes in human colonic mucosae.     

Intraductal Papilloma of the Male Breast: An Ultrastructural and Immunohistochemical Study

A case of intraductal papilloma of the male breast was studied by electron microscopy and immunohistochemistry. The major components of this lesion were the epithelial and myoepithelial cells. Intermediate cells showing ultrastructural features of both cell types were also observed. Squamous metaplasia was noted in many areas. Numerous intranuclear helioid inclusions were seen in the tumor cells. The features of this lesion are similar to those of papillomas of the female breast.     

Intraductal Papilloma of the Male Breast: An Ultrastructural and Immunohistochemical Study

A case of intraductal papilloma of the male breast was studied by electron microscopy and immunohistochemistry. The major components of this lesion were the epithelial and myoepithelial cells. Intermediate cells showing ultrastructural features of both cell types were also observed. Squamous metaplasia was noted in many areas. Numerous intranuclear helioid inclusions were seen in the tumor cells. The features of this lesion are similar to those of papillomas of the female breast.     

So-Called Glandular Schwannoma: Ependymal Differentiation in a Case

A cervical root tumor in a patient with neurofibromatosis showed a biphasic pattern of spindle and epithelioid cells with prominent “gland” formation, characteristic of the so-called glandular schwannoma. Electron microscopy and histochemistry of the “glands” disclosed features consistent with an ependymal differentiation. It is noted that there is a curious preferential association of ependymal lesions and neurofibromatosis, the pathogenesis of which is not understood.     

So-Called Glandular Schwannoma: Ependymal Differentiation in a Case

A cervical root tumor in a patient with neurofibromatosis showed a biphasic pattern of spindle and epithelioid cells with prominent “gland” formation, characteristic of the so-called glandular schwannoma. Electron microscopy and histochemistry of the “glands” disclosed features consistent with an ependymal differentiation. It is noted that there is a curious preferential association of ependymal lesions and neurofibromatosis, the pathogenesis of which is not understood.     

Superficial Acral Fibromyxoma: Immunohistochemical and Ultrastructural Analysis of a Case,with Literature Review

ABSTRACT

Superficial acral fibromyxoma (SAFM) is an uncommon tumor of the superficial soft tissues of acral sites. SAFM is a proliferation of fibroblastic cells, within a myxoid to collagenous stroma. The published cases mostly expressed immunoreactivity for CD34, CD99, EMA, and, less frequently, CD10. The authors report an additional case that did not express any of the previously reported markers, including CD34, and antigens of mesenchymal stromal lineage. Ultrastructural study confirmed the tumor cells were typical fibroblasts with cytoplasmic intermediate filaments and numerous cisternae of rough endoplasmic reticulum. The authors describe the first example of SAFM, ultrastructurally studied, with pure fibroblastic immunoprofile.     

Sclerosing Stromal Tumor of the Ovary: An Ultrastructural and Immunohistochemical Analysis with Histogenetic Considerations

Sclerosing stromal tumors are rare, benign ovarian neoplasms of unknown etiology and histogenesis. Three sclerosing stromal tumors were evaluated by immunohistochemistry and electron microscopy and were compared to two thecomas and nonneo-plastic ovarian mesenchymal tissue. The sclerosing stromal tumors and thecomas were positive for muscle-specific actin; immunoreactivity was intense in the cellular areas of the sclerosing stromal tumors and focal in the thecomas. This antigen was expressed in nonneoplastic stroma predominantly in a perifollicular (theca external distribution. Two sclerosing stromal tumors and both thecomas were vimentin positive. Desmin was present in nonvascular cells in one of each tumor type. Expression of vimentin diffusely and of desmin focally was present in nonneoplastic cortical stroma and surrounding follicles. All specimens were nonreac-tive for cytokeratin. Electron microscopy supported differentiation toward smooth muscle in the sclerosing stromal tumors but not in the thecomas. Such differentiation included aggregates of cyto-plasmic filaments with interspersed dense bodies, pinocytotic vesicles, and basal lamina. Delicate, long processes interconnected cells, often with primitive junctions, in the hypocellular foci. Cyto-plasmic lipid, which was present in the thecomas, was not well developed in the sclerosing stromal tumors. It is proposed that a population of muscle-specific actin-positive elements exists in the theca externa—the perifollicular myoid stromal cell —and that sclerosing stromal tumors may originate from them. Sclerosing stromal tumors and thecomas share many antigenic determinants and morphologic features and thus are probably closely related entities.     

Sclerosing Stromal Tumor of the Ovary: An Ultrastructural and Immunohistochemical Analysis with Histogenetic Considerations

Sclerosing stromal tumors are rare, benign ovarian neoplasms of unknown etiology and histogenesis. Three sclerosing stromal tumors were evaluated by immunohistochemistry and electron microscopy and were compared to two thecomas and nonneo-plastic ovarian mesenchymal tissue. The sclerosing stromal tumors and thecomas were positive for muscle-specific actin; immunoreactivity was intense in the cellular areas of the sclerosing stromal tumors and focal in the thecomas. This antigen was expressed in nonneoplastic stroma predominantly in a perifollicular (theca external distribution. Two sclerosing stromal tumors and both thecomas were vimentin positive. Desmin was present in nonvascular cells in one of each tumor type. Expression of vimentin diffusely and of desmin focally was present in nonneoplastic cortical stroma and surrounding follicles. All specimens were nonreac-tive for cytokeratin. Electron microscopy supported differentiation toward smooth muscle in the sclerosing stromal tumors but not in the thecomas. Such differentiation included aggregates of cyto-plasmic filaments with interspersed dense bodies, pinocytotic vesicles, and basal lamina. Delicate, long processes interconnected cells, often with primitive junctions, in the hypocellular foci. Cyto-plasmic lipid, which was present in the thecomas, was not well developed in the sclerosing stromal tumors. It is proposed that a population of muscle-specific actin-positive elements exists in the theca externa—the perifollicular myoid stromal cell —and that sclerosing stromal tumors may originate from them. Sclerosing stromal tumors and thecomas share many antigenic determinants and morphologic features and thus are probably closely related entities.     

Variability of Differentiation Patterns in Xenotransplanted Spindle Cell Sarcomas: A Histomorphological, Immunohistochemical, and Ultrastructural Study

Three leiomyosarcomas, 3 nerve sheath sarcomas, 1 rhabdomyosarcoma, and 1 sarcoma not otherwise classifiable with 17 of their xenografts, grown on nude mice, were analyzed to assess the degree of concordance between histomorphology, immunohistochemistry, and ultrastructure in spindle cell sarcoma xenograft differentiation. Histomorphology was inconclusive or misleading in 4/8 sarcoma strains and immunohistochemistry in 4/8 originals and in 10/17 xenografts, although specific patterns had been identified ultrastructurally. Electron microscopy was superior to immunohistochemistry and histomorphology in spindle cell sarcoma differential diagnosis. A further purpose of this study was to clarify whether spindle cell sarcoma xenografts retain the morphological characteristics of their primaries. Histomorphological features of the primaries were preserved over all passages, whereas the immunohistochemical marker profiles as well as the ultrastructural phenotypes changed in 14/17 xenografts and in 8/17 xenografts, respectively. Moreover, unusual bidirectional or tridirectional patterns of differentiation were identified ultrastructurally with leiomyomatous as well as Schwann cells occurring side by side and with MFH-like areas in 5/17 xenotransplants. These findings suggest genetic instability of tumor cells and may be important in the consideration of mesenchymal differentiation pathways.