Myoepithelioma of the parotid gland initially diagnosed by fine needle aspiration cytology and immunocytochemistry: a case report

BACKGROUND: Myoepithelioma is a rare, benign tumor of the salivary gland, most commonly affecting the parotid gland. Although the cytologic features of myoepithelioma are documented in a few case reports, it has rarely been diagnosed preoperatively by fine needle aspiration (FNA) cytology. CASE: A 33-year-old man presented with a left parotid swelling 2.5 cm in diameter and of about 5 years' duration. FNA smears showed bundles of spindle-shaped cells as well as plasmacytoid and stellate cells in sheets and dissociated forms. A few cells had nuclear grooves, and occasional cells showed intranuclear cytoplasmic inclusions. In May-Grünwald-Giemsa-stained smears, most of the cells had reddish cytoplasm. Red to purple, myxoid matrix was present as a scanty fibrillar substance and as globules surrounded by tumor cells vaguely reminiscent of adenoid cystic carcinoma. A cytodiagnosis of myoepithelioma was given and corroborated by immunocytochemical staining, which revealed a positive reaction for vimentin, smooth muscle actin and S-100 protein. Epithelial membrane antigen yielded a negative reaction except for a few plasmacytoid cells with weakly positive staining. Histopathology of the resected tumor and immunohistochemical staining confirmed the cytodiagnosis of myoepithelioma. CONCLUSION: FNA cytologic features together with immunocytochemical studies on smears can offer a preoperative diagnosis of myoepithelioma.     

Cytomorphologic and immunocytochemical studies of chlamydial infections in cervical smears

A total of 872 cells in 183 Papanicolaou-stained cervical smears morphologically suspected of harboring chlamydial infections were cytologically investigated in an attempt to differentiate the morphologic features of chlamydial infection from those of mucus vacuoles or bacterial infection. The observed inclusions were classified according to their morphologic appearance and their staining by a Chlamydia-specific peroxidase-antiperoxidase stain and by the periodic acid-Schiff technique. Immunoperoxidase-positive inclusions were detected in 201 cells from 13 cases; 200 (99.5%) of these cells contained "nebular" inclusions while 1 cell (0.5%) contained multiple inclusions with homogeneous central target formation. These findings suggest that nebular-type inclusions may be the key morphologic finding for the identification of chlamydial infection and that the application of an immunoperoxidase staining technique on the destained Papanicolaou preparation may be useful for the diagnosis of equivocal inclusions.     

MEMBRANOUS STRUCTURES IN YEASTS

1. Most yeast cells carrying out active respiration have spherical or ellipsoidal mitochondria, with plate-like cristae. 2. Cytoplasmic petite strains of Saccharomyces cerevisiae have aberrant mitochondria, often containing whorled membranes. Mutants with deficiencies in the tricarboxylic acid cycle have mitochondria which appear normal when the cells are grown in low levels of glucose. 3. Cells of normal and petite S. cerevisiae grown strictly anaerobically show no recognizable mitochondrial profiles. 4. Carbon substrates which can only be respired promote the development of well-defined mitochondria. In certain facultatively anaerobic yeasts respiration is suppressed by glucose and the mitochondria under these conditions are large, pleomorphic and few in number. Other fermentable carbohydrates do not give this repression. 5. A number of antibacterial antibiotics, which inhibit mitochondrial protein synthesis, cause a disorganization of the mitochondrial cristae. 6. In yeast cells adapting from anaerobic to aerobic conditions mitochondria appear to develop from proliferations of the endoplasmic reticulum, which become progressively more organized. 7. Vacuoles often contain granular material, but in S. cerevisiae the vacuole, which has been described as a lysosome, frequently contains myelin-like lipid inclusions. The material in these inclusions is apparently derived from spherosomes. 8. Endoplasmic reticulum, orientated parallel to the plasmalemma, may be associated with fermentative ability in certain facultatively anaerobic yeasts. Endoplasmic reticulum is also actively involved in the budding process. 9. Normally the yeast-cell plasmalemma shows only minor convolutions, but in chloramphenicol-grown Rhodotorula glutinis the plasmalemma produces vesicular structures termed ‘paramural bodies’. 10. The yeast nuclear membrane has about 200 pores occupying 6–8 % of the total surface area. The nuclear membrane remains intact during mitotic division in yeasts until the daughter nuclei separate.     

Adult rat cardiomyocytes cultured in creatine-deficient medium display large mitochondria with paracrystalline inclusions, enriched for creatine kinase

In adult regenerating cardiomyocytes in culture, in contrast to fetal cells, mitochondrial creatine kinase (Mi-CK) was expressed. In the same cell, two populations of mitochondria, differing in shape, in distribution within the cell and in content of Mi-CK, could be distinguished. Immunofluorescence studies using antibodies against Mi-CK revealed a characteristic staining pattern for the two types of mitochondria: giant, mostly cylindrically shaped, and, as shown by confocal laser light microscopy, randomly distributed mitochondria exhibited a strong signal for Mi-CK, whereas small, "normal" mitochondria, localized in rows between myofibrils, gave a much weaker signal. Transmission EM of the giant mitochondria demonstrated paracrystalline inclusions located between cristae membranes. Immunogold labeling with anti-Mi-CK antibodies revealed a specific decoration of these inclusions for Mi-CK. Addition of 20 mM creatine, the substrate of Mi-CK, to the essentially creatine-free culture medium caused the disappearance of the giant cylindrically shaped mitochondria as well as of the paracrystalline inclusions, accompanied by an increase of the intracellular level of total creatine. Replacement of creatine in the medium by the creatine analogue and competitor beta-guanidinopropionic acid caused the reappearance of the enlarged mitochondria. It is believed that the accumulation of Mi-CK within the paracrystalline inclusions, similar to those observed in certain myopathies, represents a compensatory effect of the cardiomyocytes to cope with a metabolic stress situation caused by low intracellular total creatine levels.     

Fine needle aspiration cytology of primary proximal-type epithelioid sarcoma of the perineum: a case report

BACKGROUND: Epithelioid sarcoma is a malignant soft tissue tumor of unknown histogenesis. We describe the cytologic findings in a case of primary proximal-type epithelioid sarcoma of the perineum and results of an immunofluorescence analysis of rhabdoid cells from this tumor. To the best of our knowledge, the 3-color immunofluorescence features of proximal-type epithelioid sarcoma have never before been reported. CASE: An 8-cm-diameter mass with a 2.5-cm ulcer was found in the perineum of a 36-year-old man. After excision of the tumor, histopathologic examination of the resected specimen suggested a diagnosis of proximal-type epithelioid sarcoma. Fine needle aspiration cytology showed numerous rhabdoid cells with globular intracytoplasmic inclusions. A few isolated cells and polygonal cells were also observed in the smears. Three-color immunofluorescence analysis indicated that the intracytoplasmic inclusions in the rhabdoid cells were positive for cytokeratin, vimentin and CD34. The cytoplasmic staining pattern differed between rhabdoid and epithelioid sarcoma cells. CONCLUSION: Immunofluorescent staining of rhabdoid cells from a primary perineal proximal-type epithelioid sarcoma revealed an uneven distribution of cytokeratin in intracytoplasmic inclusions, with the highest concentration at the periphery of the inclusions.     

Histochemical observations on the juxtanuclear complex of organelles in the hamster oocyte.

The hamster ovarian oocyte surrounded by 2-3 layers of granulosa cells develops a large juxtanuclear complex of organelles, consisting of basophilic substance, mitochondria and some lipid inclusion bodies. The basophilic substance staining for RNA, protein and some diffuse lipoprotein forms the material with which the granular mitochondria, composed of protein and phospholipid, apparently multiply. The lipid inclusion bodies consist of phospholipids. The results of this histochemical investigation have been correlated with those of ultrastructural studies on the hamster oocyte. The paranuclear complex of organelles described in this paper has also been compared and contrasted with that seen in the primordial oocyte of hamster, as reported in the previous publication. The functional significance of juxtanuclear entities has been discussed in relation to the multiplication of organelles to be accumulated during oocyte growth.     

On degeneration of peptidergic neurosecretory fibres in the albino rat

Three types of degenerating peptidergic neurosecretory fibres have been found in the posterior pituitary of chronically dehydrated albino rats. "Dark" neurosecretory fibres and their swellings contain neurosecretory granules, neurotubules, shrunken mitochondria and diffusely distributed fine dense material. Some swellings are filled with synaptic vesicles and/or conglomerations of dense membranes. The transitional forms exist between these fibres and extracellular accumulations of electron dense material. Synaptic vesicles, single neurosecretory granules, lipid-like droplets and lamellar bodies occur in the latter. Some neurosecretory fibres and swellings have numerous polymorphous inclusions arising due to degradation of secretory inclusions and organelles, mitochondria and neurotubules in particular. "Dark" neurosecretory elements and those with numerous polymorphous inclusions are enveloped by pituicyte cytoplasm. Sometimes the plasma membranes both of the pituicytes and neurosecretory fibres are destroyed or transformed into a multi-membrane complex. It is assumed that pituicytes may phagocytize degenerating neurosecretory elements. N urosecretory fibres with a locally dissolved neuroplasm and/or large lucent vacuoles seem to be due to axonal degeneration by the "light" type. These neurosecretory elements, the largest of them in particular, may transform into large cavities bordered by a membrane and containing flake-like material and single-membrane vacuoles. Degeneration of neurosecretory elements seems to occur mainly due to hyperfunction of the hypothalamo-hypophysial neurosecretory system.     

An ultrastructural assessment of mitochondria in the gastric parietal cell with the high iron cliamine method

Summary The ultrastructural densification of mitochondria by the high iron diamine (HID) method has been investigated by staining gastric parietal cells with each component of the HID reagent, alone or in combination, or with an otherwise modified HID solution. The effect of certain chemical treatments prior to staining has also been assessed. These tests provided evidence for at east three cytochemically distinguishable constituents in the mitochondrial matrix. In addition, the results from these tests and observations obtained with a newly introduced diaminobenzidine (DAB)-FeCl₃ staining reagent indicated that the probable mode of action of the HID method in imparting density to mitochondrial matrix entails combination between a complex of iron with polymerized or aggregatedm- andp-diamine and some matrix constituent. Assessment of the cytochemical staining in varied fixation condition revealed that the lucent population of mitochondria recently described in HID-stained parietal cells reflects a failure to stain with the method because of inadequate fixation.The possible nature of the HID-reactive substance in the mitochondrial matrix is discussed in the light of these observations and other cytochemical reactivities.     

蚕豆萎焉病毒2感染豌豆叶细胞后引起的线粒体增生和聚集

应用超薄切片和免疫金标记电镜技术,结合体视学分析研究了受蚕豆萎焉病毒2(BBWV 2) 中国分离物B935侵染的豌豆(Pisum sativum)叶细胞中线粒体的异常变化。结果表明,感病细胞线粒体增生并聚集于细胞质的膜增生区周围,体积增大,形状畸变,一些线粒体内含有类结晶包涵体。病叶细胞与健康对照之间线粒体的体积密度(VV)、表面积密度(SV)、数密度(NV)等参数存在显著差异(P<0.01),而形状因子(PE)、周长指数(CI)、比表面积(RSV)等参数随不同病变阶段而有变化。在线粒体周围及线粒体之间的网格结构可被BBWV 2金标记抗体特异性标记．推断为正在组装的病毒粒子。子代病毒形成结晶体和管状体,有高密度的免疫金颗粒标记。上述研究结果提示BBWV 2 引起的细胞线粒体异常变化与病毒复制组装有关,聚集线粒体的外膜粘连面可能是病毒粒子组装部位,一些线粒体内的类结晶包涵体可能代表了某种蛋白质异常积累。     

Two types of intracytoplasmic inclusions in the cells of a patient with acute lymphoblastic leukemia

A patient with acute lymphoblastic leukemia (ALL) with cells containing two types of cytoplasmic inclusions is described. The inclusions appeared as globular bodies containing electron dense material with homogeneous structure and as crystalloid formations confined in organelles with structure similar to that of the surrounding mitochondria. In distinction to other reports, these structures were not related to the endoplasmic reticulum. The possibility that some of them represented altered mitochondria is discussed.     

Origin of nucleolus-like bodies found in the nucleoplasm and cytoplasm of Vicia faba meristematic cells

Cytohistochemical staining and RNase-gold labelling have been applied to root-tip meristematic cells of Vicia faba to study the origin and biological significance of 2 types of inclusions: one seen in the nucleoplasm and the other in the cytoplasm of early telophase cells. They have been termed "dense bodies" and "cytoplasmic nucleolus-like bodies" (NLB), respectively. Both types of inclusions respond positively to silver staining and ribonucleoprotein (RNP) staining in a similar fashion to nucleolus. Interestingly, the dense bodies label heavily with the RNase-gold complex, as does the nucleolus, while the cytoplasmic NLB have no affinity with the label. In most cases, the dense bodies label more heavily than the nucleolus. Light microscope surveys reveal that the dense bodies sometimes appear to be released from the surface of the nucleolus. On the other hand, prenucleolar material showing the same silver staining and RNP preferential staining characteristics as the dense bodies begin to accumulate on the surface of chromosomes in mid-anaphase. This material does not label with RNase-gold. These data are discussed in terms of the hypothesis that the dense bodies are derived from the nucleolus by direct budding or fragmentation, and the cytoplasmic NLB are composed of prenucleolar material that failed to attach to chromosomes.     

Intramitochondrial filamentous bodies in the thick limb of henle of the rat kidney

"Intramitochondrial filamentous bodies" (IMFB) were occasionally found within the matrix of some mitochondria of the thick limb of Henle of the rat kidney, but not elsewhere in the tubular system. Three types were recognized: type I, an accumulation of filaments 55 A thick; type II, a bundle of parallel filaments having the same thickness as those of type I and regular spacing, 87 A apart, from center to center; and type III, consisting of type II with regular light bands of 280 A periodicity and a helical border of prismatic tubular cristae. In addition to these, electron-opaque masses showing variable and faint substructures were found in the matrix of mitochondria. It is suggested that all these IMFB may originate from mitochondrial cristae and that type II IMFB may be an intermediate developmental form between type I and type III. After uranyl acetate staining, IMFB and the membranes of prismatic tubular cristae showed highly increased electron opacity. The literature has been reviewed for reports of intramitochondrial filamentous inclusions in various types of cells. These inclusions have been classified according to their structural characteristics and the localization in the mitochondria and compared with IMFB reported herein.     

An immunogold-silver staining method for detection of cell surface antigens in cell smears

We developed an indirect immunogold-silver staining method for detection of leukocyte cell surface antigens in cell smears. Air-dried and fixed cytocentrifuge preparations or smears of peripheral blood leukocytes were incubated with monoclonal antibodies (MAb) and colloidal gold-labeled secondary antibodies. The preparations were post-fixed and silver enhancement was performed. The smears were counterstained with May-Grunwald-Giemsa and examined in brightfield light microscopy. The morphology of the cells was well preserved. Leukocytes reacting with the MAb showed black granules on their surface membranes. The intense immunostaining and the low background allowed a rapid enumeration of the positive cells. The labeling could be detected with high sensitivity by epipolarization microscopy. This immunogold-silver staining method was used to quantify T- and B-lymphocytes and natural killer cells in buffy coat smears of normal adult blood. These lymphocyte subsets correlated well with those obtained in smears with the alkaline phosphatase-anti-alkaline phosphatase (APAAP) method and with those found by labeling of mononuclear cells in suspension with immunogold-silver staining. This immunogold-silver staining method forms a good alternative to immunoenzyme methods for study of hematologic cells. In addition, it could be a general procedure for detection of cell surface antigens in all kinds of cell smears.     

蚕豆萎焉病毒2感染豌豆叶细胞后引起的线粒体增生和聚集

应用超薄切片和免疫金标记电镜技术,结合体视学分析研究了受蚕豆萎焉病毒2（BBWV2）中国分离物B935侵染的豌豆（Pisumsativum）叶细胞中线粒体的异常变化。结果表明,感病细胞线粒体增生并聚集于细胞质的膜增生区周围。体积增大,形状畸变,一些线粒体内含有类结晶包涵体。病叶细胞与健康对照之间线粒体的体积密度（Vv）、表面积密度（Sv）、数密度（Nv）等参数存在显著差异（P〈0．01）,而形状因子（PE）、周长指数（CI）、比表面积（Rsv）等参数随不同病变阶段而有变化。在线粒体周围及线粒体之间的网格结构可被BBWV2金标记抗体特异性标记。推断为正在组装的病毒粒子。子代病毒形成结晶体和管状体,有高密度的免疫金颗粒标记。上述研究结果提示BBWV2引起的细胞线粒体异常变化与病毒复制组装有关。聚集线粒体的外膜粘连面可能是病毒粒子组装部位。一些线粒体内的类结晶包涵体可能代表了某种蛋白质异常积累。     

The ultrastructure of the osmeterium and the nature of its secretion in Papilio larvae (lepidoptera)

Two main cell types constitute the defensive osmeterium gland of Papilio larvae. Ellipsoid gland cells have an extensively infolded basal plasma membrane, abundant ribosomes and whorls of smooth endoplasmic reticulum. The apical plasma membrane bears long microvilli extending into a mass of granular material containing electron-lucid cavities. Tangential slits occur in the epicuticle. Tubular arm cells contain heterogeneous, electron-dense inclusions, extensively-branched nuclei and large mitochondria sometimes distended with electron-dense material. The apical plasma membrane bears short microvilli. The inner, dense epicuticle forms a complex ramifying system. The two-phase defensive fluid consists mainly of water, 2-methyl propionic acid, and 2-methyl butyric acid.     

Fine Structure of Cellular Inclusions in Measles Virus Infections

Cells which are infected with measles virus have been known for some time to contain inclusion material that is distinguishable from normal cellular components by application of traditional staining methods and observation in the light microscope. The fine structure of the inclusion material contained in HeLa cells infected with Edmonston strain of measles virus has been examined in the electron microscope. Two steps have been found necessary in this study: (1) the recognition by phase-contrast microscopy of the living cell of bodies that are defined as inclusion material when the cells are classically stained; and (2) the recognition in the electron microscope of inclusion-body material that had previously been identified in the living cell. The fine structure of the nuclear and cytoplasmic inclusion material in osmium-treated cells was found to consist mainly of randomly arrayed filaments of low electron density. Dense, highly ordered arrays of filaments were found near the center of the nuclear inclusions, sometimes as a two-dimensional, nearly orthogonal arrangement. If the size of the measles virus is taken to be around 100 mµ in diameter, the strands seen in the inclusions cannot be fully formed virus.     

Ultrastructure of crystalloid inclusions in the dog and rat epididymis

Fine structural studies of the epididymis of mature mongrel dogs and of Sprague-Dawley rats were undertaken in conjunction with research dealing with the effects of vasectomy upon this organ. This paper reports the observation of crystalloid and lamellar inclusions present in these species following fixation of the epididymis in 5 % glutaraldehyde, post-fixation in osmium, and routine processing for electron microscopy. In the dog, crystalloid inclusions were observed within the cauda epididymidis of unoperated and vasectomized animals. They were found within the apical cytoplasm of principal cells in association with the Golgi apparatus and endoplasmic reticulum, and in some instances, in close proximity to the nucleus. These crystalloids exhibited a 12 nm periodicity and often measured over 3 μm in length. In the rat, two types of inclusions were found, one within mitochondria of clear cells from unoperated animals and another within membrane-bound bodies of principal cells from the caput epididymidis of unoperated and vasectomized animals. The mitochondria which contained inclusions were basally located and were observed in stacks of up to eight elongate mitochondria each. The mitochondrial inclusions exhibited a complex lamellar structure with an approximate periodicity of 36 nm. In contrast, the crystalloid inclusions found within principal cells were sequestered within supranuclear cytoplasmic bodies which increased in number with age. Such crystalloids exhibited a linear periodicity of 11–13.5 nm, but the precise lattice structure remains to be determined. Although certain aspects of the morphology of these bodies suggests a relationship to microbodies, we have been unable to demonstrate catalase activity within them. At present, neither the origin of crystalloid structures described, nor their relationship to epididymal physiology is clear.     

Occurrence of Mitochondria in the Nuclei of Tobacco Sperm Cells

Tobacco sperm cells contain intact mitochondria within their nuclei with a frequency of 0.35 [plusmn] 0.13 per cell. These inclusions appear to originate from mitochondria found among chromatids in the highly elongated metaphase plate of the dividing generative cell. These organelles are apparently captured during the reconstitution of the nuclear envelope. Only sperm cells were observed to contain these nuclear mitochondria; generative cells, vegetative pollen cells, transmitting tissue cells, unfertilized egg cells, and central cells lacked them. Nuclear mitochondria were also seen in the nuclei of the egg and central cell after fusion with sperm nuclei, suggesting that nuclear mitochondria are transmitted into the zygote and primary endosperm cells during double fertilization. Organellar inclusions in the sperm nucleus provide a potential mechanism for transmitting organellar DNA into the next generation and could potentially facilitate the transfer of genetic material between the nucleus and other organelles.     

Solitary fibrous tumour: a diagnostic challenge for the cytopathologist

N. Gupta, A. Barwad, K. Katamuthu, A. Rajwanshi, B. D. Radotra, R. Nijhawan and P. Dey Solitary fibrous tumour: a diagnostic challenge for the cytopathologist Background: Solitary fibrous tumour (SFT) is an uncommon spindle cell tumour that can occur in a variety of locations. Cytological features of this tumour have only rarely been reported in the literature. We describe the cytomorphological features of SFT with an emphasis on diagnostic pitfalls. Methods: We retrieved nine cases of histopathologically proven SFT. Three cases had sampling error with inadequate smears and, therefore, six cases with adequate cellularity were analysed for cytological findings. The cytomorphological features and the differential diagnoses on fine needle aspiration cytology (FNAC) are discussed. Results: No definitive cyto‐diagnosis of any of these cases was possible because of the morphological overlap with various soft tissue tumours and other tumour types. There was one false‐positive case, in which the possibility of sarcoma was suggested due to the presence of scattered atypical cells. Cytologically, the smears from the SFTs showed spindle to plump cells embedded in metachromatically staining dense ropy collagen material. The cells usually had oval to spindle shaped nuclei, bland chromatin and wavy elongated pale staining cytoplasm. Conclusion: A diagnosis of SFT on cytology smears is challenging. Careful attention given to certain cytological features in an appropriate clinicoradiological setting and application of immunochemistry, including CD34 and CD99 immunostaining on cytological samples, can help in the diagnosis of SFT in some cases. It is important to consider cytological overlaps of this tumour in order to avoid false‐negative or false‐positive results.     

Fibrillar bodies in leukemic cells revealed by polarization microscopy.

An optical polarizing microscope with a good coefficient of extinction permits the visualization of the cytoplasmic fibrillar body in living preparations and smears of leukemic cells (human leukemias and the L 5222 experimental leukemia). These inclusions are not visible by phase contrast microscopy nor in fixed and stained smears. The detection in living cells of fibrillar bodies makes it possible to study directly the conditions for their formation and their reaction to the effect of certain drugs.