Carcinosarcomas of the esophagus

Two cases of carcinosarcoma of the esophagus are reported. Both were polypoid tumors occurring in the middle of the intrathoracic esophagus. The tumors were predominantly composed of spindle-shaped sarcoma cells with some squamous cell carcinomas (SCC). One tumor showed many bizarre giant cells with filamentous materials in the cytoplasm. Microscopical examination of both tumors revealed transition from SCC to sarcoma cells. Immunohistochemi-cally, the spindle-shaped sarcoma cells in both tumors displayed a strongly positive immunoreaction to alpha-smooth muscle actin, as did the bizarre giant cells of one tumor to sarcomeric actin. SCC and a few spindle-shaped sarcoma cells near the SCC showed a positive immunoreaction to cytokeratin. Electron microscopy revealed that the spindle-shaped cells had many myofilaments with dense bodies and that the bizarre giant cells had sarcomere structures with 2-bands in their cytoplasm. These findings indicate that both tumors were carcinosarcomas of SCC and myogenic sarcoma. We considered that sarcoma cells might originate in SCC, representing its metaplastic differentiation, or that both SCC and sarcoma might originate in a pluripotent stem cell.     

Cadherin expression in the developing chicken cochlea.

In this study, we demonstrate that eight classic cadherins are differentially expressed in distinct anatomical regions of the cochlea during late stages of chicken embryonic development. Cadherin-6B is expressed in hair cells and spindle-shaped cells, while cadherin-8 mRNA is found only in supporting cells. Cadherin-11 is widely expressed not only in mesenchymal cell around the cochlea, but also in supporting cells and homogene cells. N-cadherin is found in the sensory epithelium, the neurons of the acoustic ganglion and on their neurites that target the hair cells. Three closely related cadherins (cadherin-7, cadherin-19, and cadherin-20) are expressed in a partially complementary manner in spindle-shaped cells and acoustic ganglion cells. R-cadherin is observed in homogene cells, acoustic ganglion cells, and their projections to hair cells. The expression of classic cadherins in the developing cochlea suggests a role for cadherins in the development of the cochlea.     

Malignant myoepithelioma of the breast (histological, immunomorphological and electron microscopic study)

A rare kind of a breast tumor was identified in a woman of 47. The disease ran a malignant course and presented difficulties for morphological verification. Histological appearance showed areas of spindle-shaped or lamellar cells occasionally encircled with homogeneous hyalin-like substance. Locally, there were structures of alveolar or glandular type with an irregular basal membrane and loose position of cells. The primary histological diagnosis suggested a tumor of mesenchymal origin. Followed by immunomorphologic (monoclonal antibody to KL 1) and electron-microscopic examinations, the diagnosis was verified providing evidence for epithelial nature of the tumor cells, loose, forming glandular-like structures and simulating stromal elements with ultrastructural features typical of myoepithelial differentiation. The evidence obtained justify the tumor definition as a rare neoplasm of the breast known in the literature as myoepithelioma.     

Detection of mRNAs for urokinase-type plasminogen activator, its receptor, and type 1 inhibitor in giant cell tumors of bone with in situ hybridization.

Although giant cell tumor of bone (GCT) is generally considered to be an uncommon benign neoplasm, it can pursue an aggressive course with local recurrence and metastasis. Attempts to predict the biological behavior of GCT with histopathological parameters, however, have not been successful. The urokinase-type plasminogen activation system has been implicated in tumor invasion and metastasis and abnormalities of the components of this system have been found in several malignancies. In this study we postulated that the urokinase-type plasminogen activation system associated with bone destruction and local invasion is present in GCT. We therefore evaluated the mRNA levels for urokinase-type plasminogen activator (u-PA), urokinase-type plasminogen activator receptor (u-PAR), and plasminogen activator inhibitor type 1 (PAI-1) by using Northern blot analysis and in situ hybridization in four cases of GCT and spindle-shaped mononuclear cells at the 35th passage from a GCT. Our results showed that giant cell tumors of bone contained variable levels of u-PA, u-PAR, and PAI-1 mRNA, respectively, 2.3, 1.4, and 3.2 kb in size. In situ hybridization showed that u-PA, u-PAR, and PAI-1 mRNA were expressed in both the mononuclear cells and the osteoclast-like giant cells; the signal for u-PA mRNA in the spindle-shaped mononuclear cells was more intense than that in the osteoclast-like multinuclear giant cells. Some spherical mononuclear cells (macrophage-like cells) expressed high levels of PAI-1 mRNA in comparison with the spindle-shaped mononuclear cells. In addition, the 35th passaged spindle-shaped mononuclear cells were used to study the gene expression of u-PA during cell proliferation. The results showed that the level of u-PA mRNA increases after adding 10% fetal calf serum to quiescent cells. The induction was maximal at 16 hours and remained high during 48 hours of treatment. In conclusion, even though osteoclast-like cells are ultimately responsible for the bone resorption of GCT, the mononuclear neoplastic cells of GCT may also be involved in degradation of the extracellular matrix during invasive growth by facilitating the urokinase plasminogen activation system. In addition, our observation of upregulation of u-PA mRNA in spindle-shaped mononuclear cells after serum stimulation indicated that u-PA production may be linked to tumor growth.     

Proliferative myositis and fasciitis. A light and electron microscopic, cytologic, DNA-cytometric and immunohistochemical study.

Two cases of proliferative myositis, four cases of proliferative fasciitis and one mixed form of proliferative myositis and fasciitis have been analyzed in terms of cell differentiation and DNA content. Light microscopically, the lesions were characterized by a mixture of proliferating spindle-shaped cells and uni-, bi- or occasionally multinucleated ganglion cell-like cells. The spindle cells showed ultrastructural features and immunohistochemical properties, including an immunoreactivity for smooth muscle-specific actin, indicative of a myofibroblastic differentiation. The ganglion cell-like cells displayed some resemblance to active osteoblasts ultrastructurally and differed immunohistochemically from the spindle cells by being non-immunoreactive for smooth muscle-specific actin. None of the two cell types showed immunoreactivity for desmin, myoglobin or factor VIII RAG. It is suggested that the two cell types represent different lines of cell differentiation. The cytologic features in smears, as seen in two cases of proliferative fasciitis and one case of proliferative myositis, are considered to be characteristic of these lesions and to permit the diagnosis to be made by fine-needle aspiration. In two of the cases, the lesion was diagnosed only cytologically and thereafter disappeared spontaneously within a month. Cytometric DNA measurements, using two different image analysis systems on Feulgen-stained sections and smears, revealed a "diploid" spindle-shaped cell population with a variable proportion of cells with scattered DNA values. The ganglion cell-like cells differed from the spindle cells by having a broad DNA peak in the diploid region and additional peaks in the tetraploid region, as well as a higher proportion of cells with scattered DNA values compared with those of the spindle-shaped cells. The results of the quantitative DNA analysis are well in keeping with the benign and proliferative nature of these lesions. However, with the technique used here, quantitative DNA analysis, does not distinguish these pseudosarcomatous fibrous lesions from diploid and tetraploid soft tissue sarcomas.     

Congenital leiomyomatous epulis: a case report with immunohistochemical study

The histologic and immunohistochemical findings of an extremely rare case of congenital soft tissue mass on the alveolar ridge in an infant are reported. The lesion clinically mimicked an ordinary congenital epulis (congenital granular cell epulis, granular cell tumor of the newborn); however, histologically it consisted of a conglomerate of spindle-shaped cells, akin to smooth muscle cells, which formed interlacing and whorled fasciculi. Nerve fibers with myxoid degeneration, capillaries and muscle walled small vessels intermingled with fasciculi of spindle-shaped cells. The border between the conglomerate of spindle-shaped cells and the surrounding connective tissue was not evident. Immunohistochemically, most of the spindle-shaped cells were intensely positive for antibodies to alpha-smooth muscle actin, HHF-35 and desmin. These findings suggest that the lesion was composed of mature smooth muscle cells that were of hamartomatous or choristomatous nature. The term 'congenital leiomyomatous epulis' is proposed.     

Cytoskeletal organization, vinculin-phosphorylation, and fibronectin expression in transformed fibroblasts with different cell morphologies

Neoplastic transformation of fibroblasts results in widely different cell morphologies. We have attempted to correlate cell morphology with cytoskeletal organization and fibronectin expression in murine and avian fibroblasts transformed by a diverse group of viral and chemical agents. The distribution of vinculin, alpha-actinin, actin, and surface fibronectin was studied, and, where appropriate, also the extent of phosphotyrosine modification of vinculin. Irrespective of the transforming agent we found that increased cell rounding was generally correlated with a reduction in vinculin-containing focal adhesions, a dissolution of microfilament bundles, and a reduction of extracellular fibronectin. In contrast, spindle-shaped fibroblasts expressed relatively high levels of surface fibronectin. Reorganization of vinculin, actin, and alpha-actinin into rosette-like structures was observed in polygonal or rounded cells transformed by viruses encoding tyrosine kinases, but was not seen in fibroblasts transformed by agents without associated tyrosine kinase activity or in spindle-shaped cells. No correlation was found between the extent of phosphotyrosine modification of vinculin and the extent of cell rounding. Irrespective of cell morphology, the extent of tyrosine phosphorylation of vinculin was high in all cells transformed by viruses carrying the src gene, but low in those transformed by viruses expressing the fps gene. Our results indicate that the morphology of a transformed cell is determined by a combination of several factors which are affected to different extents by different transforming agents.     

Development of spindle-shaped cells and chondroid cells from androgen-dependent Shionogi carcinoma 115. A light and electron microscopic study

Androgen-dependent Shionogi carcinoma 115 (SC115) is an undifferentiated medullary carcinoma consisting of compact round cells. However, when host male DS mice were castrated 2 weeks after tumor transplantation, tumors composed of compact round cells, spindle-shaped cells and chondroid cells grew 4 weeks after castration. Compact round cells with desmosomes were arranged in solid nests and exhibited immunoreactivity for keratin protein. Spindle-shaped cells had prominent rough endoplasmic reticulum, and appeared to secrete collagen. Chondroid cells had the characteristics of chondrocytes. The light and electron microscopic features were highly suggestive of a transition from compact round cells to spindle-shaped cells, and from spindle-shaped cells to chondroid cells. The histology of this tumor thus suggests that SC115 cells are able to change into chondroid cells via spindle-shaped cells.     

Sarcomatoid carcinoma of the pancreas: A case report with immunohistochemical study

Sarcomatoid carcinoma of the pancreas is an uncommon neoplasm. The immunohistochemical characteristics of this unique type of pancreatic tumor were studied. Histologically, there was diffuse proliferation of atypical spindle cells that had hyperchromatic, short, spindle-shaped nuclei and pale cytoplasm. A few tiny foci of small tubular structures were seen in connection with the atypical spindle-shaped cells. Immunohistochemical examination showed that the spindle cells were positive for epithelial cell markers (cytokeratin AE3, cytokeratin AE1, epithelial membrane antigen) and DF3 (MUC1 apomucin-related antigen (ARA)), and were negative for markers such as vimentin, desmin, neuron-specific enolase, and myoglobin. DF3 antigen is known to be expressed in invasive ductal carcinoma of the pancreas and liver, as well as of the breast. Other MUC1-ARA (MY.1E12, MUC1 glycoprotein, HMFG-1, HMFG-2) and anti-CA19-9 were also detected in the present case. Thus, this tumor was diagnosed as anaplastic carcinoma (sarcomatoid carcinoma).     

DEVELOPMENT OF SPINDLE-SHAPED CELLS AND CHONDROID CELLS FROM ANDROGEN-DEPENDENT SHIONOGI CARCINOMA 115: A Light and Electron Microscopic Study

Androgen-dependent Shionogi carcinoma 115 (SC115) is an undifferentiated medullary carcinoma consisting of compact round cells. However, when host male DS mice were castrated 2 weeks after tumor transplantation, tumors composed of compact round cells, spindle-shaped cells and chondroid cells grew 4 weeks after castration. Compact round cells with desmosomes were arranged in solid nests and exhibited immunoreactivity for keratin protein. Spindle-shaped cells had prominent rough endoplasmic reticulum, and appeared to secrete collagen. Chondroid cells had the characteristics of chon-drocytes. The light and electron microscopic features were highly suggestive of a transition from compact round cells to spindle-shaped cells, and from spindle-shaped cells to chondroid cells. The histology of this tumor thus suggests that SC115 cells are able to change into chondroid cells via spindle-shaped cells. ACTA PATHOL JPN 38: 1405–1416, 1988.     

Spindle cell carcinoma of the larynx

The clinicopathological features of 14 laryngeal neoplasms consisting of spindle-shaped cells are presented. Light microscopy showed a variety of morphological patterns from that of pleomorphic sarcoma and fibrosarcoma to more loose vascular patterns. Immunohistochemistry revealed that the spindle-shaped cells had a positive keratin immunoreactivity in 8 of 14 cases, and also that some cases had a dual expression of keratin and vimentin filaments. Electron microscopy showed that spindle-shaped cells had epithelial ultrastructures. The results support the hypothesis that the spindle cell carcinomas are true carcinomas with mesenchymal metaplasia and that the spindle-shaped cells are part of the neoplasm and not benign, reactive fibroblasts. These lesions occurred mainly on the true vocal cords in elderly patients. The neoplasms were nearly all polypoid, and many also ulcerated. There is no significant difference in clinical behaviour between laryngeal spindle cell carcinomas and ordinary squamous cell carcinomas of the larynx, and the same treatment policy is therefore advocated. Being polypoid and therefore able to be surgically removed with relative ease, they may even present a more favourable clinical course.     

Examination of crystalline arrays in non-a,non-b hepatitis

Individual particles within crystalline arrays found in the endothelial cells of hepatic sinusoids in experimental non-A, non-B hepatitis were examined by the Markham rotation technique. The particles appeared to possess an outer structure with 16–18 divisions. The presence of the crystalline structures is probably a reflection of host cell response to infection.     

Imprint cytology of primary cardiac sarcomas: a report of 3 cases

Primary cardiac sarcomas are rare instances and only occasionally documented in the cytologic literature. Usually, the diagnosis of these rare lesions can be made at echocardiography, aspiration biopsy cytology, cardiac biopsy, and open cardiac surgery (intraoperative diagnosis). In this study, cytologic configurations and immunohistochemistry for 3 primary cardiac sarcomas (rhabdomyosarcoma, angiosarcoma, and malignant fibrous histiocytoma) were revealed. In rhabdomyosarcoma (right ventricle), the tumor cells exhibited an anisocytotic spindle-shaped nuclei with hyperchromasia and an obscure cytoplasmic margin. Vimentin and myosin were positive throughout the cytoplasm for the tumor cells. In angiosarcoma (right atrium), small clusters of anisocytotic spindle-shaped tumor cells appeared as vascular-like structures and hemosiderin-laden macrophages in many erythrocyte-rich backgrounds. Nuclei showed round to oval shape with hyperchromasia and prominent large nucleoli. Cytoplasm was obscure and elongated. Factor VIII related antigen and CD34 were strongly positive throughout the cytoplasm for the tumor cells. In malignant fibrous histiocytoma (right ventricle), the tumor cells exhibited oval to spindle-shaped and elongated nuclei and coarse granular chromatins with hyperchromasia. The nuclear margin was thin. A few small round nucleoli appeared. Elongated obscure and foamy cytoplasm was stained pale blue. Vimentin and α₁-antitrypsin were positive throughout the cytoplasm for the tumor cells. This study elucidated the cellular characteristics and immunohistochemistry for cardiac sarcomas using imprint smears as an aid to cytopathologic diagnosis.     

Punch and spindle-shaped biopsies for collecting oral mucosal tissue for the fabrication of transplantable autologous epithelial cell sheets

The oral mucosa is an easily accessible source of cells. Oral mucosal collection will be an essential surgical procedure for regenerative medicine and cell biological research. However, there is no current report that describes the details of the surgical procedure used for oral mucosal collection. Moreover, the number of cells that can be obtained has not been determined. Two different procedures, the punch biopsy and the spindle-shaped biopsy, were performed for the fabrication of transplantable autologous epithelial cell sheets. The mean values of the cells collected per square centimeter of tissue using the punch biopsy and the spindle-shaped biopsy were 76.8 ± 45 × 10(4) cells/cm(2) and 195.7 ± 120 × 10(4) cells/cm(2) , respectively. There was no significant difference between the punch biopsy and the spindle-shaped biopsy. The coefficient of variation of the punch biopsy and the spindle-shaped biopsy was 58.9% and 69.8%, respectively. This result indicated that both procedures showed variations in the number of collected cells. Although the punch biopsy may be easier and simpler than the spindle-shaped biopsy, multiple punch biopsies may result in a more complicated procedure, and the spindle-shaped biopsy may be preferable when a large number of cells is necessary. ? 2012 Wiley Periodicals, Inc. J Biomed Mater Res Part A 100A:2849-2854, 2012.     

大鼠骨髓源性内皮祖细胞的分离培养与鉴定

背景：内皮祖细胞因其分离与培养的方法各不相同,在实验中难以重复。 目的：探讨大量获取骨髓源性内皮祖细胞分离与培养的方法。 方法：通过密度梯度离心法从4周龄SD大鼠骨髓中分离单个核细胞,使用EGM-2 MV培养基进行诱导培养,采用形态学特征观察、摄取Dil-Ac-LDL与结合FITC-UEA-1实验、免疫荧光化学鉴定其表面抗原CD133与VEGFR2等方法对其进行鉴定,并通过管腔形成实验观察形成管腔的能力。 结果与结论：①形态学观察：分离的骨髓单个核细胞经诱导培养后,在生长的早期(8 d左右)、晚期(15 d左右)其细胞形态有一定差异,早期以纺锤形、三角形、圆形细胞多见,晚期以圆形、短梭形细胞多见。②摄取Dil-Ac-LDL与结合FITC-UEA-1实验：显示8,21 d的细胞均为阳性。③免疫荧光化学染色：8 d的细胞表达CD133、VEGFR2。④管腔形成实验：在Matrigel基质上15 h左右能够生成血管样结构。结果表明：利用密度梯度离心法分离大鼠骨髓单个核细胞后以EGM-2 MV进行诱导培养,经过鉴定证明获得的细胞符合内皮祖细胞的特征。这种方法能够简单、快速、可靠、大量地获取内皮祖细胞。     

Coronavirus infection of rat dorsal root ganglia: ultrastructural characterization of viral replication, transfer, and the early response of satellite cells

Swine hemagglutinating encephalomyelitis virus (HEV) has been shown to have a capability to gain access to the cell bodies of sensory neurons after peripheral inoculation, resulting in ganglionic infection. It is not clearly understood how this virus is replicated within and released from the sensory neurons, and it remains to know how satellite cells response to the HEV invasion. By ultrastructurally examining HEV-infected rat dorsal root ganglia, we found that HEV in the cell bodies of infected neurons budded from endoplasmic reticulum-Golgi intermediate compartments, and were assembled either individually within small vesicles or in groups within large vesicles. The progeny virions were released from the sensory neurons mainly by smooth-surfaced vesicle-mediated secretory pathway, which occurred predominantly at the perikaryal projections and infoldings of sensory neurons. Released HEV particles were subsequently taken up by the adjacent satellite cells. Almost all virus particles in the cytoplasm of satellite cells were contained in groups within vesicles and lysosome-like structures, suggesting that these glial cells may restrict the local diffusion of HEV. These observations give some insights into the pathogenesis of coronavirus infection and are thought to help understand the interactions between sensory neurons and their satellite cells.     

Diffuse malignant pericardial mesothelioma

A 67-year-old male case of pericardial mesothelioma is reported. The diagnosis was established by the gross appearance at autopsy and its histological findings. The tumor arising from the anterior epicardium of the right half of the heart was located within the pericardial sac, and displaced and compressed the heart to the posterior direction. Histologically, large areas of the tumor were composed of spindle-shaped malignant cells which were arranged in interlacing bundles. The other areas showed the presence of tubular structures and ill-defined solid nests. Distant metastasis was found only in a small focus of the 8th vertebral bone marrow composed of spindle-shaped cells. A meticulous search failed to find any tumor in other organs. Based on these findings, the tumor was diagnosed as diffuse malignant mesothelioma of biphasic type.     

Distribution of calcitonin gene-related peptide in the rat peripheral nervous system with reference to its coexistence with substance P

This immunocytochemical study, using a double-staining method, showed that calcitonin gene-related peptide-like immunoreactive structures are widely distributed in the peripheral nervous system and that many of them coexist with substance P-like immunoreactive structures in single sensory ganglion cells. Neurons positive for calcitonin gene-related peptide but negative for substance P were detected in sensory ganglia. These cells were large (about 30-45 micron in diameter); these primary sensory neurons containing calcitonin gene-related peptide can probably act independently of substance P. There were neurons containing calcitonin gene-related peptide without substance P in the pterygopalatine ganglion, although these cells were less numerous than in the sensory ganglia. In consecutive sections, calcitonin gene-related peptide-like structures occurred in thyroid parafollicular cells, which also contain calcitonin. This suggested that messenger RNA for producing calcitonin gene-related peptide is also present in the thyroid, and like calcitonin, calcitonin gene-related peptide may have a peripheral physiological role.     

Ultrastructure of hepatic angiosarcoma in rats induced by vinyl chloride

The hepatic angiosarcoma studied was from a male Wistar rat having been exposed to vinyl chloride monomer (VCM) by the oral route for 120 weeks. Widened sinusoids lined by large electron-lucent, spindle-shaped tumor cells and membrane-bound nuclear-free structures were seen in the transitional zone between the tumor mass and the adjacent liver tissue. In areas merely consisting of tumor tissue the tumor cells had a more angular or irregular shape. The origin of the tumor cells is discussed in light of the characteristic differences between endothelial cells and Kupffer cells described in the literature. Although an endothelial origin of the tumor cells could not be established unequivocally, it was concluded that at present there is little reason to doubt such an origin.