Ultrastructural identification of a proximal tubulopathy without crystals in a relapsed multiple myeloma patient

A multiple myeloma patient, who had been treated with a hematopoietic stem cell transplant, underwent a renal biopsy for investigation of a possible relapse of disease as indicated by increased serum creatinine and positive urinary Bence-Jones protein containing increased kappa light chain. Paraprotein-related renal disease was not evident by light microscopy or immunofluorescence microscopy however electron microscopy demonstrated a proximal tubulopathy with intracytoplasmic non-crystalline inclusions. The ultrastructural findings suggested possible end-organ involvement by the disease and follow-up studies subsequently revealed a relapsed multiple myeloma in the patient. The case exemplifies the usefulness of electron microscopy in detecting paraproteins that, in some instances, may be difficult to demonstrate by other techniques.     

Extraskeletal myxoid chondrosarcoma: a light microscopic, immunohistochemical, ultrastructural and immuno-ultrastructural study indicating neuroendocrine differentiation

AIMS: Extraskeletal myxoid chondrosarcoma is a rare low-grade soft-tissue sarcoma with locally aggressive and metastasizing potential. Extraskeletal myxoid chondrosarcoma has distinctive clinical, light microscopic, immunophenotypic, cytogenetic and ultrastructural features. Evidence that extraskeletal myxoid chondrosarcoma often shows neuroendocrine features was first provided by Chhieng et al. on the basis of an immunohistochemical and ultrastructural study of seven cases. Our study aims to further confirm by immunohistochemistry and ultrastructural studies, including immunoelectron microscopy, that extraskeletal myxoid chondrosarcoma indeed may show neuroendocrine differentiation. METHODS AND RESULTS: Fifteen cases of extraskeletal myxoid chondrosarcoma and seven control cases of skeletal chondrosarcomas were studied. Extensive immunohistochemical analysis was performed in all cases and ultrastructural studies were done in 11 extraskeletal myxoid chondrosarcomas and three skeletal chondrosarcomas. Immunoelectron microscopy was performed on one case each of extraskeletal myxoid chondrosarcoma and skeletal chondrosarcoma. Extraskeletal myxoid chondrosarcomas expressed neuron-specific enolase (100%), synaptophysin (87%), S100 (50%), PGP 9.5 (40%), and epithelial membrane antigen (25%). Co-expression of synaptophysin and PGP 9.5 was observed in six tumours. Skeletal chondrosarcomas showed expression of S100 protein, vimentin and neuron-specific enolase in all cases. Synaptophysin, chromogranin and PGP 9.5 were not expressed in any skeletal chondrosarcoma case. Ultrastructurally, extraskeletal myxoid chondrosarcoma was characterized by distinct cords of cells immersed in a glycosaminoglycan-rich matrix. The cells were rich in mitochondria, had well-developed Golgi apparatus and there were numerous smooth vesicles. In three cases there were easily found 140-180 nm diameter membrane-bound dense-core granules in cell bodies and in processes, unrelated to the Golgi, compatible with neurosecretory granules. Fewer such granules were present in the remaining extraskeletal myxoid chondrosarcoma cases, three of which also contained intracisternal tubules typical of extraskeletal myxoid chondrosarcoma. The skeletal chondrosarcomas had scalloped cell surfaces, prominent rough endoplasmic reticulum focally distended with secretory product, and lacked neurosecretory granules. Intermediate filaments were prominent in both extraskeletal myxoid chondrosarcoma and skeletal chondrosarcomas. Immunoelectron microscopy showed synaptophysin expression in the extraskeletal myxoid chondrosarcoma but not in the skeletal chondrosarcoma case. CONCLUSIONS: This study confirms that a substantial proportion of extraskeletal myxoid chondrosarcomas show immunophenotypic and/or ultrastructural evidence of neuroendocrine differentiation, and are unlikely to be related to conventional skeletal chondrosarcomas.     

Rhabdoid tumour of the bladder: a histological, ultrastructural and immunohistochemical study

A rapidly fatal bladder tumour which had the features of a rhabdoid tumour was studied by sequential biopsies and at autopsy. This is the first rhabdoid tumour recorded at this site and the first in which there was co-existent transitional cell carcinoma. The possibility that rhabdoid tumour is histogenetically heterogeneous is discussed.     

Oval cells in hepatitis B virus-positive and hepatitis C virus-positive liver cirrhosis: histological and ultrastructural study

AIMS: It is still not clear whether oval cells demonstrate diverse morphology, immunophenotype or quantity in different human liver diseases. The aim of this study was to investigate these differences in hepatitis B virus (HBV)-positive and hepatitis C virus (HCV)-positive human liver cirrhosis (HLC). METHODS AND RESULTS: Thirty-eight cases of HBV+ HLC and 32 cases of HCV+ HLC were investigated by light microscopy and immunohistochemistry for Hepatocyte, CK19, stem cell factor (SCF) and CD34. Five cases were also examined by transmission electron microscopy. Oval cells of similar morphology could be found in proliferating bile ductules in both groups. These cells coexpressed CK19 and Hepatocyte, but did not express SCF or CD34. Some of these cells exhibited a trend towards differentiation. There was no difference in the amount of oval cells between the two groups. The oval cell number was found to increase significantly with the progression of inflammation. A similar stem-like cell was not seen in the normal liver. CONCLUSIONS: There are bipotential oval cells in both HBV+ and HCV+ HLC. The lack of difference in oval cells between the two groups suggests that they might play a similar biological role in the histogenesis of different liver diseases.     

Malignant haemangiopericytoma of lung: a cell culture and ultrastructural study

Malignant haemangiopericytoma was grown in vitro to assess the histogenesis of the tumour. Disaggregated tumour cells maintained in serum-supplemented Waymouth's MB 752/1 medium showed monolayers of homogeneous spindle cells. Post-confluent cultures exhibited hillocks or plaques of multilayer growth within which considerable amount of basal lamina-like material and fibrillar matrix were present between cells. Delicate basal laminae were also expressed on cell surfaces facing matrix material. Other consistent features were attenuated cytoplasmic processes, desmosome-like junctions, abnormal mitochondria and a paucity of intracellular filaments. The three-dimensional organization of tumour cells with concomitant expression of differentiated phenotype in vitro has significant implications on the cell of origin and differentiation process of haemangiopericytoma.     

Proximal tubular cells contain a phenotypically distinct,scattered cell population involved in tubular regeneration

Regeneration of injured tubular cells occurs after acute tubular necrosis primarily from intrinsic renal cells. This may occur from a pre‐existing intratubular stem/progenitor cell population or from any surviving proximal tubular cell. In this study, we characterize a CD24‐, CD133‐, and vimentin‐positive subpopulation of cells scattered throughout the proximal tubule in normal human kidney. Compared to adjacent ‘normal’ proximal tubular cells, these CD24‐positive cells contained less cytoplasm, fewer mitochondria, and no brush border. In addition, 49 marker proteins are described that are expressed within the proximal tubules in a similar scattered pattern. For eight of these markers, we confirmed co‐localization with CD24. In human biopsies of patients with acute tubular necrosis (ATN), the number of CD24‐positive tubular cells was increased. In both normal human kidneys and the ATN biopsies, around 85% of proliferating cells were CD24‐positive – indicating that this cell population participates in tubular regeneration. In healthy rat kidneys, the novel cell subpopulation was absent. However, upon unilateral ureteral obstruction (UUO), the novel cell population was detected in significant amounts in the injured kidney. In summary, in human renal biopsies, the CD24‐positive cells represent tubular cells with a deviant phenotype, characterized by a distinct morphology and marker expression. After acute tubular injury, these cells become more numerous. In healthy rat kidneys, these cells are not detectable, whereas after UUO, they appeared de novo – arguing against the notion that these cells represent a pre‐existing progenitor cell population. Our data indicate rather that these cells represent transiently dedifferentiated tubular cells involved in regeneration. Copyright © 2012 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.     

恶性颗粒细胞瘤临床病理、免疫组化和超微结构观察

目的：探讨恶性颗粒细胞瘤的病理学诊断和鉴别诊断要点及组织学起源。方法：对３例恶性颗粒细胞瘤进行临床病理、免疫组化及超微结构观察研究。结果：男性２例，女性１例，平均年龄为４９岁。部位分别为颈部１例，右大腿２例。其中２例分别于术后２年半及７年复发，并伴有区域淋巴结转移。组织学上３例与良性颗粒细胞瘤十分相似，局部区域出现梭形瘤细胞，空泡状核及明显的核仁，其中１例在肿瘤的周边部可见到瘤细胞与外周神经束支之间有直接移行关系。免疫酶标结果显示瘤细胞强阳性表达Ｓ－１００蛋白和神经特异性烯醇化酶（ＮＳＥ），１例电镜检测显示胞浆内充满膜包被复合性溶酶体。结论：对临床明显恶性而组织学上却极似良性的恶性颗粒细胞瘤，以下几点能提示恶性诊断：（１）肿瘤超过４ｃｍ；（２）核分裂象超过２个／１０ＨＰＦ；（３）核呈空泡状并有明显的核仁；（４）出现梭形瘤细胞；（５）有肿瘤性坏死。此外，免疫组化标记及超微结构观察有助于鉴别诊断及揭示组织学起源。     

Large-cell calcifying Sertoli cell tumor of the testis: a case report with ultrastructural study

Large-cell calcifying Sertoli cell tumor (LCCSCT) is a rare histologic variant of Sertoli cell tumor. Recently we observed a case of LCCSCT of the testis with no associated endocrine abnormality. Our ultrastructural findings of rows of tight junctions, numerous intracytoplasmic filaments, and abundant rough endoplasmic reticulum in whorled formations support the Sertoli cell origin of this neoplasm.     

Cystadenoma of the pancreas: a histological, histochemical and ultrastructural study of seven cases

Seven cases of cystadenoma of the pancreas were examined in detail histologically and histochemically and two variants were identified: a microcyst type lined by glycogen-rich cuboidal epithelium which also secretes some neutral mucins; and macrocystic type lined exclusively by mucus-secreting columnar cells which secrete predominantly sulphated acidic mucins with some neutral mucins. Electron microscopy of two cases of the microcystic variant showed neoplastic cells with an ultrastructure comparable to that of centroacinar cells of the normal exocrine pancreas.     

Malignant histiocytosis: a histological and ultrastructural study of lymph nodes in six cases

A morphological, histological, and ultrastructural investigation was carried out on a series of six cases of malignant histiocytosis observed between 1973 and 1977. All lymph nodes were obtained prior to treatment. The histological findings revealed that the proliferating cell type was in all cases represented by histiocytes of varying degrees of atypia and with consistent phagocytic activity. In four cases, the histiocytic cellular proliferation also showed some degree of cohesiveness and, in two cases, there was capsular invasion. In three cases blood vessel invasion by malignant cells could be seen within the lymph node and in the surrounding tissue. On electron microscopy the tumour cells of malignant histiocytosis appeared to be pleomorphic with three types of cells: undifferentiated cells, histiocytes with variable degrees of differentiation, and cells with intermediate features. In particular, nonphagocytic and actively phagocytic histiocytes could be identified. The histological and ultrastructural data further support the idea that malignant histiocytosis is a disease that is related to the neoplastic proliferation of moderately differentiated histiocytes and their precursors. The latter may undergo various transformations with either apparently benign or definitely atypical morphological features.     

Benign papillary mesothelioma of peritoneum: a histological, histochemical and ultrastructural study of six cases

Six cases of localized benign papillary mesothelioma of peritoneum were retrieved from laboratory files over a 20-year period. One presented because of pain following torsion and the remaining five were incidental findings at operation. Light microscopy showed a delicate papillary stroma covered by a single layer of mesothelial cells. The mesothelial cells often showed punctate periodic acid-Schiff staining of cytoplasm. The brush border and stroma showed alcian blue positive staining which was abrogated by prior incubation with hyaluronidase. Electron microscopy revealed typical features of mesothelial cells with long microvilli, rough endoplasmic reticulum in relation to mitochondria, bundles of microfilaments, and tight junctions. Between cells there was a series of intercommunicating channels containing occasional lymphocytes and debris. These features are compared with nodular mesothelial hyperplasia, malignant mesothelioma and adenomatoid tumours of the genital tract. The necessity to distinguish benign papillary mesothelioma from metastatic carcinoma is stressed and relies on the recognition of their mesothelial origin.     

Paraganglioma of the cauda equina: a histological, ultrastructural and immunocytochemical study of two cases with a review of the literature

The clinical and pathological features of two paragangliomas arising in the cauda equina are described and compared with 14 previous reports. The light microscopic features were similar to those of paragangliomas from other sites, with a 'Zellballen' pattern of cells containing argyrophil granules. Electron microscopy showed densely staining membrane-bound granules, cilia and fibrous bodies in the cytoplasm. The last two features only occur in paragangliomas from this site. gamma-Enolase was demonstrated by the peroxidase-antiperoxidase technique for the first time in these neoplasms. This technique was also used to demonstrate cytokeratins, which appear to be associated with the presence of fibrous bodies. The pathological findings suggest that paragangliomas in this site arise from pre-existing paraganglia, possibly of the visceral autonomic group. The prognosis in cases treated by complete excision appears to be good.     

Immunogold labelling of PCNA and Ki-67 antigen at the ultrastructural level in laryngeal squamous cell carcinoma and its correlation with lymph node metastasis and histological grade

Streptavidin-gold was used for the immunolocalization of PCNA and Ki-67 antigen at the ultrastructural level with a postembedding technique in biopsies of 15 patients with laryngeal squamous cell carcinoma. Positive immunoelectron staining was obtained in 9 cases for PCNA (60%) and in 8 cases for Ki-67 (53%). PCNA was predominantly found in heterochromatin of the nucleus of laryngeal carcinoma cells in a granular pattern. Positivity for PCNA was not found in nucleoli. In 4 cases, positive staining was observed both in nucleus and cytoplasm. In the cytoplasm, it was found to be present on the endoplasmic reticulum and on ribosomes throughout the cytoplasm. Ki-67 antigen was localized in the nucleus where it was associated with heterochromatin and euchromatin. It was also observed in nucleoli in all cases. Cytoplasmic localization of Ki-67 antigen was similar to that of PCNA. All 8 cases that were positive for Ki-67 were also positive for PCNA. Control incubations did not result in labelling with steptavidin-gold particles for both antigens. A significant correlation between PCNA and Ki-67 expression in association with pathological characteristics such as nodal status and histological grade was not found. Our data indicate that Ki-67 antigen staining correlates with PCNA labelling, whereas a relationship between proliferation markers and tumour progression was not found.     

Clear cell papillary renal cell carcinoma: a clinicopathological study emphasizing ultrastructural features and cytogenetic heterogeneity

Clear cell papillary renal cell carcinoma (CCPRCC) is a recently recognized renal neoplasm, which was initially described in end-stage renal disease (ESRD), but some cases have been reported in otherwise normal kidneys. We report a series of 11 CCPRCC (age range, 33-72 years; male-to-female ratio, 8:3). Follow-up was available for 8 patients. No patients developed local recurrence, distant or lymph-node metastasis, or cancer death. Histologically, all tumors exhibit morphologic features typical of CCPRCC including a mixture of cystic and papillary components, covered by small to medium-sized cuboidal cells with abundant clear cytoplasm. All 11 cases exhibited moderate to strong positivity for CK7, CA9, Vim, and HIF-1α, coupled with negative reactions for CD10, P504S, and RCC. We did not find any VHL gene mutations in all 11 cases. Losses of chromosomes 3 (monoploid chromosome 3) was detected in 3 cases. Ultrastructurally, the tumor cells composed of numerous glycogens with scanty cell organelles, reminiscent of clear cell renal cell carcinoma (CCRCC). In conclusion, the coexpression of CA9 and HIF-1α in the absence of VHL gene abnormalities in CCPRCC suggests activation of the HIF pathway by mechanisms independent of VHL gene mutation. Losses of chromosomes 3 (monosomies chromosome 3) was detected in 3 cases suggesting that at least some of these lesions have demonstrated abnormalities of chromosomes 3. Ultrastructurally, CCPRCC composed of numerous glycogens with scanty cell organelles, reminiscent of CCRCC suggesting the close pathogenesis relationship of CCPRCC with CCRCC.     

Human malignant mesothelial cells: Variability of ultrastructural features in established and nude mice transplanted cell lines

The aim of this study was to determine, by transmission electron microscopy, the differentiation features of 21 human malignant mesothelioma cell lines (HMCLs) established from 13 specimens of 12 confirmed human malignant mesotheliomas, and of tumours induced in nude mice injected with 16 HMCLs. Fifty per cent of HMCLs showed typical mesothelial differentiation (long and slender microvilli, desmosomes, perinuclear intermediate filaments); 29 per cent did not show differentiation; and the remainder were poorly differentiated. Three human tumour specimens gave several different HMCLs; the cell lines obtained from a given tumour exhibited variable mesothelial differentiation. Eleven HMCLs were compared with the native tumour. Four were similar to the tumour and seven were less well differentiated, in most cases in relation to their microvilli. With six HMCLs, tumours induced in nude mice were less well differentiated than the corresponding cell lines, whereas with four HMCLs, tumours were equally or better differentiated. However, in most nude mice tumours, typical mesothelial microvilli were present. These results show that cell lines established from malignant mesothelioma may exhibit dedifferentiated features. However, while the variability in ultrastructural differentiation may result from the culture microenvironment, it could also be related to the state of differentiation, of the native tumour sample and to tumour cell heterogeneity.     

Mesothelioma with clear cell features: an ultrastructural and immunohistochemical study of 20 cases

Mesotheliomas with clear cell morphology are rare and only a few individual case reports have been documented in the literature. The author reports a series of 20 epithelioid mesotheliomas with clear features, 17 of which originated in the pleura and 3 in the peritoneum. Eighteen of the patients were men and 2 were women. Twelve patients had a history of asbestos exposure. Electron microscopy and special histochemical stains demonstrated that the cytoplasmic clearing seen in hematoxylin and eosin-stained sections resulted from multiple factors that can occur either singly or in combination. The most frequent cause of the cytoplasmic clearing was the accumulation of large amounts of intracytoplasmic glycogen. Another but somewhat less common factor was the accumulation of large amounts of lipid, which occurred alone or with glycogen. Other less common causes were marked mitochondrial swelling, the presence of numerous intracytoplasmic vesicles, and a large number of intracytoplasmic lumens. The value of immunohistochemistry in helping to distinguish epithelioid mesotheliomas from some carcinomas with clear cell morphology is emphasized. In addition, it was determined that because electron microscopy was decisive in establishing the cause of the cytoplasmic clearing in most of the cases, tissue for electron microscopy should routinely be procured for ultrastructural studies.     

Biphenotypic plasma cell myeloma: two cases of plasma cell neoplasm with a coexpression of kappa and lambda light chains

Plasma cell neoplasm (PCM) is a medullary and extra medullary proliferation of clonal plasma cells that occurs due to accidental translocation of proto-oncogenes into immunoglobulin (Ig) gene loci. While the majority of plasma cell neoplasms are monoclonal, up to 2% of the PCMs [1] considered being biclonal based on electrophoretic analysis, characterized by secretion of paraprotein with two distinct heavy chains or light chains are possible and present unique diagnostic challenges. Methods: Traditionally protein electrophoresis has been used to diagnose, characterize, and monitor progression of plasma cell neoplasm. To characterize neoplastic plasma cells, in our institution, other ancillary studies, including in situ hybridization, flow cytometric analyses of plasma cell surface markers and cytoplasmic immunoglobulins with DNA ploidy, are also utilized routinely. Results: We present two cases of plasma cell myeloma in which the neoplastic plasma cells shows production of cytoplasmic kappa and lambda light chain, with secretion of free lambda light chain only. Co-expression of kappa and lambda light chain by the same neoplastic plasma cells is a rare but reported phenomenon. Conclusions: Our study indicates that serum electrophoresis alone could mischaracterize biphenotypic myeloma as monotypic plasma cell myelomas in the absence of additional testing methods.     

Squamous cell characteristics in small cell carcinoma of the uterine cervix: histological, immunohistochemical, and ultrastructural study in xenografted small cell carcinoma

Small cell carcinoma of the uterine cervix is a rare type of gynecological tumor that frequently expresses neuroendocrine differentiation. Its histological origin is unclear. We examined the histopathological characteristics of small cell cervical carcinoma in a patient with elevated serum adrenocorticotropin hormone. We then studied the morphological alteration in xenotransplanted tumors (passages 1–9) using immunohistochemistry and electron microscopy. The primary cervical tumor was characterized by a sheetlike arrangement of uniform small cells with hyperchromatic nuclei and a high nucleocytoplasmic ratio. A ribbon-like or trabecular pattern was also observed in a small area of the tumor. Neuron-specific enolase, chromogranin A, and S-100 were positive for the tumor cells, but cytokeratin was negative. Dense-core granules were detected by electron microscopy. In the xenografted tumor, a serial change from squamous cells to round-to-oval cells was observed. Cytokeratin was immunohistochemically stained in the squamous tumor cells but not in the other tumor cells. In contrast, chromogranin A was stained in some of the round-to-oval cells. Basal lamina underlaid the squamous tumor cells, and desmosome-like junctions were apparent. The cytoplasm was filled with well-differentiated organelles including electron-dense tonofilaments. Elliptical tumor cells resembled the primary carcinoma ultrastructurally. These findings suggest that small cell cervical carcinoma with neuroendocrine properties shares the characteristics of squamous cell carcinoma.This study was presented at the 24th Annual Meeting of the Clinical Electron Microscopy Society of Japan, Okayama, September 17, 1992.