An ultrastructural study of nonfunctioning adrenocortical adenoma

Nonfunctioning adrenocortical adenomas obtained from 40 and 60-year-old males were examined by electron microscopy. Light microscopically, the tumors were shown to consist of clear cells and compact cells in both cases. However, compact cells predominated in the former case, and clear cells in the latter. Electron microscopy revealed that most of the cells in the former case had abundant organelles, including well-developed, smooth and rough endoplasmic reticulum, Golgi apparatus, and mitochondria with elaborate tubulovesicular cristae. The findings resembled those of adenomas with Cushing's syndrome. The latter case revealed a majority of cells with numerous, large lipid vacuoles, moderate amounts of smooth endoplasmic reticulum, Golgi apparatus, and mitochondria with sparse lamellar cristae resembling features of aldosteronoma. These ultrastructural findings suggest that even nonfunctioning adrenocortical adenomas, like functioning adrenocortical adenomas, have some possibility of producing adrenal hormones. This study was presented in part at the 25th Annual Meeting of the Clinical Electron Microscopy Society of Japan, Matsumoto, September 28–30, 1993.     

Fine structure of adrenal medullary grafts in the pain modulatory regions of the rat periaqueductal gray

Summary Recent findings in our laboratory indicate that adrenal medullary grafts produce significant alterations in pain sensitivity. Electron microscopic studies were undertaken to correlate these behavioral changes with the neural interactions of the host and graft tissue in the periaqueductal gray. A striking change found 8 weeks after transplantation is that pronounced myelination has taken place both in the graft and in the host tissue. The new myelin formation in the graft has the typical appearance of PNS myelination and, in the host the appearance of CNS myelination. The endothelial cells of the capillaries in the grafted tissue are attenuated and fenestrated in contrast to those of the surrounding parenchymal tissue of the host. By 8 weeks, the graft becomes heavily encapsulated with collagen, while the host CNS tissue develops layers of glial processes outlining the graft. However, collagen and glial layers apparently do not form an absolute barrier to either cellular or humoral interaction between the host and graft tissue. Chromaffin cells can be found protruding into the host CNS tissue and sometimes forming synapses with presumably the host neuronal processes. Grafted chromaffin cells may participate as both postsynaptic and, less often, as presynaptic components of synaptic junctions. The behavioral relevance of these synaptic contacts is unclear, since similar implants of adrenal medullary tissue into the dorsal spinal cord subarachnoid space, which also induce potent analgesia, do not contain synapses. Thus, it is more likely that behavioral changes are brought about by diffusion of neuroactive substances from grafted chromaffin cells to host receptors.     

An ultrastructural study of malignant fibrous histiocytoma of the larynx

Malignant fibrous histiocytoma (MFH) of the vocal cord occurring in a 46-year-old male was studied immunohistochemically and ultrastructurally. The tumor consisted of the two main areas, pleomorphic and storiform. Immunohistochemical staining was strongly positive for α1-antitrypsin and α1-antichymotrypsin, and negative for desmin, keratin, myoglobin, actin, carcinoembryonic antigen, and S100 protein. Ultrastructurally, this MFH consisted of four different types of cells which were either fibroblast-like, histiocyte-like, myofibroblast-like, or with features of both the fibroblast and histiocyte types. Transition forms between the four types of cells were also found. This suggests that cells of MFH are derived from the same undifferentiated stem cells.     

Spontaneous atherosclerosis: An ultrastructural study in the White Carneau pigeon

Summary The atherosclerotic lesions, associated with the celiac intimal smooth muscle cushions, of four and five year old White Carneau pigeons were studied with the light and electron miscroscopes. Light microscopic examination of the spontaneous lesions demonstrated large intimal cushions composed of smooth muscle, abundant collagen, clusters of foam cells and cholesterol crystal clefts.Ultrastructural examination of the intimal atheroma revealed dilatations between apposing endothelial cells which contained a flocculent material, similar to that seen in the subendothelial space. The subendothelial compartment contained abundant collagen, extracellular lipid, vesiculated material and cell processes which contained a flocculent matrix and tubular-like elements. In addition, fibroblast-like interlaminar cells were often observed. Numerous intimal smooth muscle cells were seen which displayed varied morphology. Abundant foam cells were also present within the intimal atheromas.The presence of atherosclerotic lesions in preexisting intimal smooth muscle cushions suggests that hemodynamic factors may be important in the progression of these spontaneous lesions. Endothelial cell dilatations may provide an important route of transport for circulating elements which may accumulate within the subendothelial space. Morphologically, it appears that the smooth muscle cells undergo modification and may represent the precursors of foam cells in this species.Supported by a General Reserach Support Grant from NIH and The University of Texas     

An ultrastructural and morphometric study of bladder tumours (I)

Summary The ultrastructure of urothelium from 4 patients with no evidence of tumour was compared with that taken from 22 patients with bladder neoplasms of different grade and stage. Two features were quantified: the percentage of reduplication of the basal lamina and the percentage of discontinuous basal lamina.Tumours showed a lower percentage of reduplicated basal laminae than normal tissues, the difference being significant in 3 out of 4 types of tumours. The difference in frequency between normal tissues and non-recurring tumours was not significant, but there was a significant difference between normals and recurring tumours.All types of tumours showed discontinuities in the basal lamina, including 80% of those staged non-invasive by light microscopy. None of the normal tissues showed these. The percentage of discontinuities seen in non-recurrent tumours was half that seen in recurrent ones, but both groups were significantly higher than normals.Loss of continuity of the basement membrane distinguishes invasive from non-invasive tumours. However, there is a high probability of these being missed by light microscopy alone. Therefore, electron microscopic studies on recurrent bladder tumours would increase the accuracy of staging and prognosis.     

An ultrastructural and morphometric study of bladder tumours (II)

Summary Bladder tissues from 3 groups of patients were examined, using the light and electron microscopes (LM and TEM). One group of patients had a history of well-differentiated papillary transitional cell carcinomas and specimens were taken from cystoscopically normal areas. In a second group frank papillary carcinoma was biopsied. Finally, patients with no history of urothelial tumours and a normal cystoscopic appearance were biopsied during investigations for various benign conditions and these served as controls.In tissues from the first two groups certain differences were seen when these were compared to the controls and the frequency of these was significant. Light microscopic examination of 0.5 toluidine blue stained sections revealed an increased number of immature, small dark cells in the superficial layer of the epithelium (P<0.001). Electron microscopic examination showed that in place of the characteristic asymmetric unit membrane of mature superficial cells, the surface was frequently covered with microvilli and the junctional complexes were often atypical. There was an increased number of abnormalities in the basal lamina (P<0.001).These features were seen in the absence of cystoscopic and light microscopic changes in three out of eight patients with a history of tumours. It is, therefore, suggested that these are the earliest detectable morphological abnormalities in the pre-neoplastic urothelium.     

Parachordoma: An ultrastructural and immunohistochemical study

A case of parachordoma of the left calf in a 19-year-old Chinese female is reported. The tumour showed multinodular growth pattern and consisted of round or oval tumour cells with abundant eosinophilic cytoplasm and myxoid matrix. Tumour cells formed small nests and sometimes showed concentric arrangement. Physaliferous-like cells and undifferentiated spindle cells were occasionally observed among the cell nests. The myxoid matrix was positive for high-iron diamine stain, indicating the presence of chondroitin 4- and 6- sulphates and keratan sulphate. Ultrastructurally, well-developed rough endoplasmic reticulum, abundant intermediate filaments, microvillous cytoplasmic processes, pinocytic vesicles, and desmosome-like junctional structures were found. Tumour cells were positive for S-100 protein and vimentin, but negative for cytokeratin, epithelial membrane antigen, carcinoembryonic antigen, and desmin. These results are consistent with the definition of parachordoma as a soft tissue neoplasm consisting of cells with histology and ultrastructure similar to those of chordoma cells but with immunohistochemistry similar to that of chondroid tumour cells.     

Neurofibromatous neuropathy: An ultrastructural study

Plexiform neurofibroma is pathognomonic of neurofibromatosis 1 (NF1). An NF1-associated peripheral neuropathy has been described in a small minority of NF1 patients but its histopathological features are poorly characterized. We report the case of a 46-year-old woman presenting with bilateral supraclavicular painful masses without other stigmata of NF1. MRI showed bilateral plexiform lesions extending from cervical roots to the elbows. Nerve conduction studies documented a sensory motor polyneuropathy. Morphometric analysis of sural nerve biopsy showed a preferential loss of large-caliber myelinated fibers with a g ratio of 0.515, and the presence of regeneration clusters. By electron microscopy, marked and diffuse endoneurial fibrosis with an altered relationship between Schwann cells (SC) and collagen fibrils was observed. Moreover both myelinating and non-myelinating SC were characterized by the presence of various cell degradation products. These changes suggest that, in neurofibromatous neuropathy, a widespread axonal atrophy and degeneration take place independently on the presence of tumoral infiltration, possibly due to an impairment in SC-axon cross talk. In this case, the coexistence of plexiform neurofibromas with a peripheral neuropathy strongly suggests a diagnosis of NF1 even without fulfillment of clinical criteria. We propose that in the presence of plexiform neurofibromas, electrophysiological studies should be performed also in asymptomatic patients, in order to detect the existence of a subclinical neuropathy.     

An ultrastructural and immunohistochemical study of central neurocytoma

A case of central neurocytoma was studied by electron microscopic and immunohistochemical methods. Immunohistochemical examinations showed the tumor cells to be positive for neuron-specific enolase, sporadically positive for synaptophysin, and negative for glial fibrillary acidic protein. The Ki-67 (MIB-1) positive index value for the tumor cells was a low 1.5%. Ultrastructurally, thin cytoplasmic processes of tumor cells showed differentiation to neuronal cells containing parallel bundles of microtubules and abortive synapses with dense core vesicles and/or clear vesicles. Microvessels composed of endothelial cells without fenestrations and tight junctions in the endothelial clefts appeared in fibrillary stromas as revealed by light microscopy. Neurosecretory granules in thin cell processes may have been secreted around microvessels.     

An ultrastructural study of meconium corpuscles in human foetal colon

Summary In human foetal colon meconium corpuscles were observed in the colonic epithelium during the stage of secondary lumina development and enlargement.Transmission electron microscopy of these specimens revealed inclusion bodies in the superficial and deeper layers of the epithelium. Many of the membrane-bounded inclusion bodies contained well-preserved organelles and some inclusions contained nuclear fragments. There was evidence of nuclear fragmentation with condensed chromatin arranged in crescentic caps. The ultrastructural observations are typical of apoptosis, a mode of cell death first described in 1972 by Kerr and colleagues.Thus, meconium corpuscles are apoptotic bodies found as a result of the deletion of healthy normal cells during the reshaping and development of organs.     

An ultrastructural and morphometric study of bladder tumours (III)

Summary Quadrant biopsies were taken at cystoscopy from 12 male patients previously diagnosed on light microscopy as having flat carcinoma in situ (CIS) of the urinary bladder. There was also material available from 3 cystectomy specimens with widespread CIS associated with papillary or solid urothelial tumours.Sections of normal ureter from kidney transplant donors and biopsies from two patients investigated for non-malignant bladder conditions servered as controls.The biopsies from 4 patients were classified as mild dysplasia of the urothelium, while those from 11 patients were categorised as CIS.Biopsies categorised as mild dysplasia on light microscopy showed an increase in the number of cells with large nuclei and nucleoli when compared to controls. The number of desmosomes was significantly reduced compared to controls, while the frequency of abnormalities of the basal lamina was increased. These features were more pronounced in the CIS group.Biopsies from the CIS group could be divided into classical and large cell CIS, the latter showing a higher frequency of ultrastructural abnormalities than the classical type.The patients diagnosed as having CIS fell into two clinical categories, the early onset and the late onset group. The five patients in the former had been diagnosed as having CIS with or without urothelial tumours elsewhere within 3 months of presentation. In the remaining four patients CIS was observed after recurring episodes of papillary or solid tumours during the previous 9 months to 20 years.The biopsies of 3 out of 5 patients with early onset CIS had been classified large cell CIS, wheras only one patient out of 4 in the late onset group came into this category. An early appearance of CIS is thought to have a worse prognosis, and it is therefore suggested that large cell CIS is a more severe form of the disease.     

An ultrastructural and immunohistochemical study of cutaneous metastasis of biliary system carcinoma

An 81-year-old Japanese man noticed red papules and subcutaneous tumors on his face and trunk about 2 months before entering our clinic. He had been suffering from biliary system cancer for about one year.Light microscopic findings: Tumor cells, which formed various-sized glands lined by columnar cells, were observed in the dermis and subcutaneous tissue.Electron microscopic findings: In tumor cells, intracytoplasmic lumina lined with microvilli were observed. Cell organelles, such as mitochondria and rough-surfaced endoplasmic reticulum, were relatively well-developed. From these findings, it seems that the characteristics of these tumor cells resemble those of the biliary system.     

Pigmented granules in functional black adenoma of the adrenal gland: A histochemical and ultrastructural study

A black adenoma of the adrenal gland was laparoscopically removed from an 61-yr-old Japanese female who had clinical and laboratory findings characteristic of Cushing’s syndrome. The tumor consisted of polygonal cells that contained numerous brown pigmented granules of various sizes by routine hematoxylin-eosin staining. The histochemical study showed that the pigment had the characteristics of lipofuscin, not of melanin or neuromelanin. Electron microscopic study revealed tumor cells with two types of pigmented granules. These results show that there might be differences in the lipid metabolism of individual tumor cells in black adenomas of adrenal; which suggests an interesting histogenesis for these granules.     

Eosinophilic fasciitis ultrastructural study of an early biopsied case

Summary A 55-year-old woman with eosinophilic fasciitis was biopsied 8 weeks after the onset of her illness. Under the electron microscope the changes were almost exclusively located in the fascia with many active fibroblasts, accumulation of protocollagen fibrils (10–50 Å diameter), elastic fibre remodelling and numerous degranulating mast cells. The inflammatory infiltrate was dense and mostly composed of lymphocytes and plasma cells, with 16% eosinophils.The connective tissue changes may be part of a healing process following microinjury of the fascia. However, large numbers of lymphocytes and plasma cells are unusual in the healing process and are more common in the cellular reaction of morphea. Nevertheless, the absence of macrophages in subcutaneous fat, together with large number of eosinophils in the fascia may be considered to be distinctive features of eosinophilic fasciitis.     

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.     

An ultrastructural study of subungual exostosis, with special reference to intranuclear inclusions

A 12-year-old Japanese boy noticed a red nodule on the distal portion of his right great toe about 2 months before entering our clinic. There was evidence of traumatic injury and infection before the appearance of the nodule. Histological examination showed the subcutaneous localization of bony spicules lined by osteoblasts. Haversian canals, which contained blood vessels and connective tissue, were also seen. Electron microscopic examination revealed normal bone structure, but, interestingly, mesenchymal cells that were localized around the tumor had intranuclear inclusions composed of filamentous bundles. From these findings, this case was diagnosed as subungual exostosis. It is suggested that the appearance of intranuclear inclusions, reported here for the first time, is related to the pathogenesis of subungual exostosis.This study was presented at the 28th annual meeting of the Clinical Electron Microscopy Society of Japan, Osaka, October 17–19, 1996     

Fine-needle aspiration cytology of spindle cell neoplasms of the adrenal gland.

Three spindle cell neoplasms were encountered in a series of 46 FNA of the adrenal performed between 1984 and 1991. These neoplasms included a recurrent undifferentiated adrenal cortical carcinoma (ACC) with a predominant spindle cell pattern, a pheochromocytoma (PC), and a metastatic desmoplastic malignant melanoma (DMM). Cytologically, the ACC was characterized by the presence of numerous microtissue fragments composed of spindle-shaped malignant cells with oval to spindle-shaped nuclei, one or more nucleoli, and bipolar cytoplasmic processes. In some areas the tumor cells were dissected by vascular channels. The background contained abundant metachromatic stroma as well as individually scattered tumor cells. The PC was composed predominantly of loosely cohesive spindle-shaped cells along with more polygonal shaped cells with delicate faintly staining granular cytoplasm. The tumor cells exhibited mild anisonucleosis. The tumor fragments were well vascularized by arborizing delicate capillary channels. The DMM was composed of microtissue fragments, interlacing fascicles and loose aggregates of spindle-shaped malignant cells with hyperchromatic nuclei, small nucleoli, and an absence of cytoplasmic pigment. In each case ancillary studies including immunocytochemistry and electron microscopy (EM) were helpful in the differential diagnosis. The ACC was negative for cytokeratins, neuron-specific enolase (NSE), and muscle-specific actin (HHF), but displayed strong positivity for vimentin as well as characteristic whorls of smooth endoplasmic reticulum by EM. The PC was positive for NSE and chromogranin with no EM performed. The DMM stained for S-100 and vimentin but was negative for HMB-45, cytokeratin, and HHF. EM examination revealed rare atypical premelanosomes.(ABSTRACT TRUNCATED AT 250 WORDS)     

An ultrastructural study of pemphigoid vegetans, characterizing the possible mechanism of vegetation

Ultrastructural changes in the skin lesions of pemphigoid vegetans were observed by transmission electron microscopy. Two significant changes were recognized at the vegetating site: (1) The basal lamina was sometimes interrupted and detached from the basal cells, protruding into the dermis to form folds. In the basal cells, hemidesmosomes were not decreased in number but tonofilament bundles were fewer than normal. Occasionally, basal cell processes extended beyond the basal lamina through its gap. These features resembled the epidermal growth factor-induced morphological changes of cultured skin. It was suspected that the abnormal attachment of the basal lamina to the basal cell was associated with keratinocyte proliferation. (2) Mast cells were encountered in the epidermis abutting on the basal lamina. It is uncommon to find mast cells in the epidermis. Mast cells in the epidermis were reported at the erythematous regions which were adjacent to the bullae of bullous pemphigoid, and were thought to have an important role in forming blisters. It was suggested that early changes in blister formation occur at the vegetating site.This study was presented at the 20th annual meeting of the Clinical Electron Microscopy Society of Japan, Fukuoka, September 1, 1988, at the 21st annual meeting of the Clinical Electron Microscopy Society of Japan, Kobe, September 22, 1989, and at the 23rd annual meeting of the Clinical Electron Microscopy Society of Japan, Toyama, September 14, 1991.     

An ultrastructural study on anterior cruciate ligament reconstruction using autogenous patellar tendon grafts in dogs

An ultrastructural study was undertaken concerning morphological changes within the autografted patellar tendon (PT) after being transplanted to the anterior cruciate ligament (ACL) in mongrel dogs. After 4 weeks, the arrangements of both large and small collagen fibrils in the PT graft became disordered, and the number of inflammatory cells increased. However, the same PT grafts revealed postsurgical signs of newly-produced collagen fibrils around activated fibroblasts at 12 to 24 weeks. At 52 weeks after the transplantation, small collagen fibrils increased in both number and density, showing a remarkable morphological similarity to the collagen fibrils of normal ACL. These data indicate that the characteristics of the PT graft eventually resemble those of a normal ACL. This paper was presented in part at the Combined Meeting of the Orthopedic Research Societies of the USA, Japan and Canada in Banff, Alberta, Canada, October, 1991.     

Ret protein expression in adrenal medullary hyperplasia and pheochromocytoma

Ret is a developmentally regulated tyrosine kinase involved in formation and maintenance of the nervous system. Ret mutations predisposing to pheochromocytomas and medullary thyroid carcinomas occur in multiple endocrine neoplasia (MEN) syndromes 2A and 2B. Biochemical studies have demonstrated overexpression of Ret mRNA and protein in pheochromocytomas compared to normal adrenal medulla. However, the cellular distribution of Ret in the normal human adrenal and in hyperplastic lesions that antecede pheochromocytomas are unclear. The present investigation was undertaken to resolve the histological distribution of Ret in the normal human adrenal, in pheochromocytomas evolving from adrenal medullary hyperplasia in MEN2A and in sporadic pheochromocytomas. Ret expression was studied by immunohistochemistry using both a polyclonal and a monoclonal antibody, with confirmation by immunoblotting of representative cases. Only occasional cells stained for Ret in the normal adrenal, consistent with the distribution in adult adrenals of other species. Heterogeneous, progressively increased Ret expression was observed during the evolution of pheochromocytomas. In both normal and neoplastic adrenal, the most intense immunoreactivity was observed in cells with neuron-like features. Our finding that Ret is not expressed at high levels in the early stages of disease suggests that elucidation of mechanisms that regulate Ret expression is required for understanding the pathobiology of MEN2A. The association of high-level Ret expression with neuronal morphology suggests that the variable overexpression of Ret in pheochromocytomas might in part be an epiphenomenon, reflecting the known phenotypic plasticity of these tumors.