Use of electron microscopy in core biopsy diagnosis of oncocytic renal tumors.

The distinction between oncocytoma and chromophobe renal cell carcinoma, important clinically, may be challenging, especially as the tissue sample size decreases. Ancillary studies can be helpful, although subject to interpretation and sample variability. The aim of this study was to examine the value of electron microscopy in differentiating between oncocytoma and chromophobe renal cell carcinoma on formalin fixed paraffin embedded needle core biopsies. Twenty renal needle core biopsies were evaluated. Despite formalin fixation and paraffin embedding, the classic ultrastructural features of these neoplasms were retained, revealing 80% sensitivity and 100% specificity by initial work-up.

A human pituitary tumor-derived folliculostellate cell line.

Pituitary cells have been used for the study of hormone synthesis, secretion, and regulation. However, the lack of human cell lines of pituitary origin has made such studies in humans very difficult. Activin, a member of the transforming growth factor-beta cytokine family, is secreted by the pituitary and serves, in addition to regulating hormone biosynthesis, as a regulator of cell growth and differentiation. In the human pituitary, folliculo-stellate cells secrete an activin-binding and -neutralizing protein, follistatin. However, the role of these cells in the autocrine/paracrine regulatory mechanisms of activin is poorly understood. We describe a human pituitary-derived folliculostellate cell line, designated PDFS, that was developed spontaneously from a clinically nonfunctioning pituitary macroadenoma. PDFS cells showed an epithelial-like morphology with long cytoplasmic processes. Electron microscopy revealed frequent intercellular junctions, including desmosomes, and cytogenetic analysis showed clonal characteristics with chromosomal abnormalities. These cells express vimentin and the nervous tissue-specific S-100 protein, specific markers of folliculostellate cells in the anterior pituitary, but no secretory pituitary cell markers. PDFS cells formed large colonies in an anchorage-independent transformation assay. They express follistatin and activin A and have an intact activin intracellular signaling pathway as determined by reporter assays. Therefore, this human cell line provides a useful model for studying the regulation of cell growth and cytokine production by factors endogenously produced in pituitary folliculostellate cells.

Hyalinizing spindle cell tumor with giant rosettes: a report of three cases with ultrastructural analysis.

We report the light microscopic, ultrastructural appearance and immunohistochemical staining profile of three distinctive soft-tissue tumors recently designated hyalinizing spindle cell tumor with giant rosettes. The tumors occurred in two men, 41 and 54 years old, and one woman, 62 years old. Two tumors arose in the lower extremities and one in the upper arm. Two tumors were resected and measured 3 and 13.2 cm in greatest diameter; a biopsy only was done of the third tumor. Grossly, the tumors had a tan, pink, or white cut surface. The largest tumor exhibited central cystic change. Microscopically, they all displayed similar features and were composed of fibromyxoid regions, with areas of hyalinization in two tumors and focal ossification in one tumor. Scattered throughout each of the tumors were rosette-like structures in which neoplastic cells were arranged around a central collagenous core. Ultrastructurally, the neoplastic cells demonstrated the features of fibroblasts. In all tumors, there was abundant extracellular collagen fibers and in one there were large aggregates of amorphous extracellular external lamina-like material. The center of the rosette-like structures was composed of banded collagen fibers and the cells at the periphery of the rosettes had ultrastructural features similar to the neoplastic spindle cells located elsewhere in the tumor. Immunohistochemically, the tumor cells stained for vimentin. There was focal staining of the widely distributed spindle cells and cells that formed the rosettes for Leu-7, S-100 protein, and CD34. In one tumor, there was faint diffuse staining of the spindle cells for neuron-specific enolase. One tumor (with the amorphous extracellular material) stained for type IV collagen. There was no staining for desmin, muscle actin, smooth muscle actin, keratin, or epithelial membrane antigen. These results demonstrate that hyalinizing spindle cell tumor with giant rosettes is composed of fibroblasts. We did not demonstrate any ultrastructural or immunohistochemical differences between the spindle cells that comprised the majority of the mass and those that surrounded the rosette-like structures. There was no ultrastructural evidence of neural differentiation to explain the focal S-100 protein and Leu-7 staining of the tumor cells.

Is anti-h-caldesmon useful for distinguishing smooth muscle and myofibroblastic tumors? An immunohistochemical study.

Misinterpretation of positive staining of antibodies to desmin, smooth muscle actin, and muscle actin as representing smooth muscle differentiation in the context of a spindle cell tumor is not uncommon. Anti-h-caldesmon is a promising novel immunohistochemical reagent for more specific smooth muscle differentiation. We studied 72 tumors (11 leiomyosarcomas, 26 malignant fibrous histiocytomas [MFHs], 11 fibromatoses, 11 cellular cutaneous fibrous histiocytomas [CCFHs], 5 malignant peripheral nerve sheath tumors, 4 synovial sarcomas, and 4 cases of nodular fasciitis), the reactive myofibroblastic response in 5 cases of acute cholecystitis, and the desmoplastic response surrounding 5 invasive breast carcinomas. Tissues were examined for expression of h-caldesmon, desmin, smooth muscle actin, and muscle actin. Diffuse staining for h-caldesmon was present only within the leiomyosarcomas. Focal staining for h-caldesmon involving less than 1% of lesional cells was present in 3 of 26 MFHs and 1 of 11 CCFHs. There was overlap in staining for the other "myoid" markers in all of the lesions that contained myofibroblasts. Anti-h-caldesmon seems to be a reliable marker of smooth muscle differentiation, and its inclusion in a panel of myoid immunohistochemical reagents should allow distinction of smooth muscle and myofibroblastic tumors.

Ultrastructure of peripheral blood granulocytes from patients with low serum cobalamin.

Peripheral blood granulocytes from ten patients with low serum cobalamin were studied using light and electron microscopic techniques. Six patients had classic neutrophil hypersegmentation using light microscopy, but three patients had an increase in band forms and rare hypersegmented cells. The electron microscope revealed nuclear appendages consisting of nuclear stalks, rings and blebs in neutrophils from all patients regardless of the degree of hypersegmentation but not from normal controls; these did not appear to result from sectioning artifact and their formation may be interrelated. Numerous cytoplasmic empty granules were seen in neutrophils from three patients. Another three showed what appeared to be granules in the shape of water-wings under low power, but on higher power were unique mitochondria. Some disorder of the eosinophil granule centrum was seen in all patients but was extreme in four patients; lipid cytoplasmic inclusions were also seen in these four and one other patient.

Apoptotic cells in peripheral blood from patients with low serum cobalamin.

Apoptotic leukocytes were found by using ultrastructural and light microscopic techniques to examine peripheral blood in ten of twelve patients with low serum cobalamin (vitamin B12) and rarely in normal controls. A total of 88 apoptotic cells (.14% of total leukocytes) were identified in all the patients. One patient also had apoptotic cells found on a routine blood smear. There was no correlation between the finding of these cells and nuclear hypersegmentation, serum cobalamin levels, serum intrinsic factor antibody, serum methylmalonic acid and homocysteine or the Schilling test. Two patients, however, with the most severe vitamin deficiency did not have increased numbers of apoptotic cells suggesting that these patients had lost the ability to initiate the cell death program.

The many faces of smooth muscle neoplasms in a gynecological sampling: an ultrastructural study.

Smooth muscle neoplasms may have a variety of light microscopic and ultrastructural appearances. On one extreme, a spindle cell mass with a fascicular pattern, located in the myometrium, usually does not need electron microscopy or immunohistochemistry to confirm its smooth muscle nature. However, at the other end of a spectrum is an epithelioid neoplasm of the extrauterine pelvic tissues that could be composed of any of several cell types if routine light microscopy, alone, were used in studying it. In this report, some of these variants of smooth muscle neoplasms are exemplified, including myxomatous, fibroblast-like, nondescript, epithelioid, granular cell, and clear cell types. The main purpose has been to address, in particular, the ultrastructure of these unusual neoplasms, but, at the same time, not to ignore or downplay the contributory role of immunohistochemistry in making a final diagnosis, in some cases. Especially intriguing were the ultrastructural characteristics of leiomyomatous granular cell and clear cell neoplasms. A paucity or absence of filaments and dense bodies in samplings of these lesions makes the reliance on other ultrastructural features extremely useful.

Chondromyxoid fibroma: a tumor showing myofibroblastic, myochondroblastic, and chondrocytic differentiation.

Chondromyxoid fibroma (CMF) is a rare primary benign tumor of bone that demonstrates variable histologic features and is often confused with chondrosarcoma. Although the chondroid elements in CMF have been reported to be S-100 protein positive and to have chondrocytic features ultrastructurally, the immunohistochemical and ultrastructural profile of CMF, especially with respect to the peripheral nonchondroid elements, has not been extensively studied. Formalin-fixed, paraffin-embedded tissue from 10 CMFs were stained immunohistochemically with antibodies to vimentin, desmin, muscle actin, smooth muscle actin, S-100 protein, and CD34. Six tumors were also examined ultrastructurally. The chondroid areas showed variable staining for S-100 protein but did not stain for muscle actin or smooth muscle actin. The peripheral areas surrounding the chondroid areas stained diffusely for smooth muscle actin and muscle actin but did not stain for S-100 protein. CD34 highlighted the extensive vascularity that was especially prominent in the peripheral areas; no tumor cells stained for CD34. There was no staining for desmin. Ultrastructural examination showed three different cell types. Some cells showed the classic features of chondrocytes, other cells had the features of myofibroblasts, and the third cell type had the features of both chondrocytes and myofibroblasts ("myochondroblasts"). These findings support the conclusion that CMF is a tumor showing myofibroblastic, myochondroblastic, and chondrocytic differentiation.