Diagnostic utility of sialosyl-Tn in discriminating carcinomatous cells from benign mesothelium in body cavity effusions

OBJECTIVE: Detecting malignant cells in the setting of reactive mesothelium can be difficult. Several techniques have been tried but without widespread acceptance. Sialosyl-Tn (STn) is an aberrantly glycosylated precursor of the MN blood group antigen frequently expressed in carcinomas and dysplastic epithelium. We investigated the STn monoclonal antibody for its clinical utility as an isolated stain to discriminate benign mesothelium from malignant cells. STUDY DESIGN: Cell block material from 72 cases of body cavity fluids were immunostained for STn using the avidin-biotin complex method without antigen retrieval. Slides were incubated overnight at 4 degrees C in a humidified chamber. RESULTS: Strong immunoreactivity was noted in 31/40 (77%) carcinomatous cases. Only moderate staining was noted in 1 of 28 (4%) benign effusions and weak staining in 5 (18%) additional benign cases. Specificity was 100%, sensitivity 78%, positive predictive value 100% and negative predictive value 76%. No staining was noted in four noncarcinomatous malignant effusions. CONCLUSION: STn may have diagnostic value in this cytologic setting as part of a diagnostic panel but not as an isolated stain.     

GLUT1 antibody staining in thin-layer specimens of benign and malignant body cavity effusions

OBJECTIVE: To determine whether GLUT1 antibody could replace one or more of the currently used antiepithelial antibodies and to assess whether ThinPrep methodology is suited to immunocytochemical (ICC) evaluation. STUDY DESIGN: In a prospective study of 10 fluids containing malignant cells from cases of proven adenocarcinoma and 10 cytologically benign effusions, multiple slides were prepared by ThinPrep technology for staining with four commercially available antibodies and appropriate isotype-matched negative controls. The antibodies used were GLUT1, CEA, B72.3 and Leu-M1 (CD 15). Tissue sections and ThinPrep slides were used as positive controls. Specimens were batched to ensure similar conditions for all antibody reactions. RESULTS: Of the 11 cases ultimately proven to be carcinoma, GLUT1 and B72.3 stained 7 each (63.6%), and CEA and Leu-M1 6 each (54.5%). No false positive staining was encountered, but one case chosen as a benign control was shown to contain immunopositive cells by three of the four epithelial markers used; this case was therefore an occult true positive rather than a false positive. CONCLUSION: In this small but controlled prospective analysis, GLUT1 demonstrated strong positive staining, with sensitivity similar to that of currently used epithelial markers. Using GLUT1 in conjunction with B72.3, no cases of carcinoma were missed. GLUT1 could be used in a panel of antibodies designed to confirm the presence of adenocarcinoma. ThinPrep methodology, which enables multiple slides to be prepared after routine microscopy determines the need for ICC, appears suited to this adjuvant investigation.     

Heparanase expression: a potential ancillary diagnostic tool for distinguishing between malignant cells and reactive mesothelium in body cavity effusions

OBJECTIVE: Heparanase, an endoglycosidase that cleaves heparan sulphate, is frequently expressed in carcinomas and was suggested to play a role in cell invasion and metastasis. We investigated whether heparanase expression may serve as a reliable marker to discriminate benign mesothelial cells from malignant cells shed into body cavities. METHODS AND RESULTS: Cytological smears of effusions from 51 hospitalized patients were immunostained for heparanase. Strong immunoreactivity was noted in 35 of 40 (88%) carcinoma samples and in all three malignant mesothelioma cases. Only rare (<3%) reactive mesothelial cells were noted showing a faint negligible staining. Specificity was 100%, sensitivity 88%, and positive and negative predictive values were 100% and 89% respectively. CONCLUSIONS: Our results suggest that heparanase may be of value as a complementary component in a diagnostic panel of markers, contributing to its reliability and accuracy.     

Comparison of light and transmission electron microscopy for the evaluation of body cavity effusions

Evaluation of 282 body cavity effusions by both light and transmission electron microscopy showed that the two methods compare favorably. The major advantages of electron microscopy are higher rates of unequivocally positive diagnoses, improved diagnosis in borderline or suspicious cases and better discrimination between benign and malignant fluids or decreased numbers of inconclusive results. This was obtained without false-positive results. The major disadvantage of the method is the increased time and preparative procedures needed. As with light microscopy, the false-negative rate was high.     

Ultrastructural and histochemical studies on guard cells

Serial thick sections of guard cells from Vicia faba L., Nicotiana tabacum L., Allium cepa L., Zea mays L. and Beta vulgaris L. were obtained systematically (600–800 nm) and viewed with the transmission electron microscope in an effort to demonstrate the presence or absence of a symplastic transport pathway within the stomatal complex. Eight to ten stomata from each species were examined, and no continuous plasmodesmata were found connecting guard cells to sister guard cells or to adjacent epidermal or subsidiary cells. Continuous plasmodesmata were observed in immature guard cells, but were sealed (truncated) during the development of the mature cell wall. Histochemical stains, phosphotungstic acid and silver methenamine, were used to demonstrate differentiation within the mature guard-cell wall. The structural differentiation of the stomatal apoplastic region is discussed in relation to fanctional specialization. Plasma-membrane elaborations or plasmalemmasomes were identified in the guard cells of Zea, and it is suggested that these structures may function in ion transport.Abbreviations PTA-HCl phosphotungstic acid and hydrochloric acid - SM silver methenamine - UA-LC uranyl acetate and lead citrate     

Ultrastructural studies of human basophils and mast cells.

Ultrastructural studies of human mast cells (HMCs) and basophils (HBs) are reviewed. Sources of HMCs include biopsies of tissue sites and in situ study of excised diseased organs; isolated, partially purified samples from excised organs; and growth-factor-stimulated mast cells that develop de novo in cultures of cord blood cells. Sources of HBs for study include partially purified peripheral blood basophils, basophils in tissue biopsies, and specific growth factor-stimulated basophils arising de novo from cord blood cells. The ultrastructural studies reviewed deal with identity, secretion, vesicles, recovery, and synthesis issues related to the biology of these similar cells.     

Ultrastructural studies on plastids of generative and vegetative cells in Liliaceae

Summary The behaviour of plastids and mitochondria during the formation and development of the male gametophyte of Chlorophytum comosum has been investigated using electron microscopy. During first pollen mitosis an intracellular polarization of plastids occurs in that the plastids are clustered in the centre of the microspore. The originating generative cell normally lacks plastids. Only in a small number of microspores have plastids been observed near the dividing nucleus of the microspore and later on in the generative cell. These observations agree with the genetic investigations of Collins (1922) on the mode of plastid inheritance which demonstrated a small amount of biparental plastid inheritance in Chlorophytum. The cytological mechanisms underlying plastid polarization during the first pollen mitosis are discussed.     

Ultrastructural identification of calbindin-D28k-immunoreactive cells in the rat superficial pineal body

The vitamin-D-dependent calcium-binding protein, calbindin-D28k, was detected immunocytochemically in the majority of pericapillary pinealocytes of the rat superficial pineal body. The interstitial cells were calbindin-negative. The significance of these findings is discussed.     

Chromosome studies on cells cultured from serous effusions. Use in routine cytologic practice

Cytogenetic analysis was used to supplement the cytologic diagnosis of malignancy in serous effusions. Current methods of analysis involve immediate preparation, but our study showed that prior culture of the cells is a more practical approach to the use of chromosome analysis in the routine cytology laboratory. This method permits the retrospective selection of specimens for chromosome analysis from patients in whom there is a strong clinical suspicion of malignant disease but who have negative or equivocal cytology reports. The preparative technique and the recognition of structural and numerical chromosome anomalies can be readily learned by the cytotechnologist and incorporated into the routine cytology laboratory.     

Ultrastructural studies in rheumatoid polyneuropathy

The biopsy material from the sural nerve of 8 patients suffering from rheumatoid arthritis and showing, with one exception, positive serological reactions, were studied by means of electron microscopy. All the patients were displaying sensory and sensomotor neuropathy. Besides hypermyelination the damage of Schwann cells, the destruction on unmyelinated fibres and the proliferation and degeneration of the vascular endothelium were observed. No inflammatory signs were seen around the peri- and endoneurial vessels.     

Ultrastructural and cytochemical studies on the cytodifferentiation of duodenal endocrine cells

Summary The development and cytodifferentiation of endocrine cells that produce the gastrointestinal hormones gastrin, cholecystokinin and secretin have been studied by a combined fluorescence-cytochemical, immunocytochemical and ultrastructural approach. The results show that, during development, several ultrastructurally distinct cell types exhibit COOH-terminal gastrin and cholecystokinin immunoreactivity. Furthermore, some cells simultaneously contain both gastrin- and cholecystokinin-specific antigenic determinants. Studies on the time course of development of gastrin and cholecystokinin cells, together with the above-mentioned data, suggest that gastrin cells may be converted into cholecystokinin cells in development. During this period, gastrin, cholecystokinin and secretin cells store the biogenic monoamine, 5-hydroxytryptamine a feature not displayed by the adult counter-parts of these cells. In the adult duodenum, characteristic enterochromaffin (EC) cells store 5-hydroxytryptamin for which, evidence for a possible hormonal role has been presented. Taken together, our data indicate that the differentiation of duodenal endocrine cells occurs in distinct steps, each involving a restriction in the biosynthetic repertoire of the cell.     

Ultrastructural studies on prolactin and growth hormone cells in Anguilla pituitaries in vitro

Summary Eel hemi-pituitaries were cultured in vitro on high or low sodium media which are known to affect differentially prolactin and growth hormone release. Ultrastructural examination of the prolactin cells after 24 h culture showed the Golgi bodies were markedly more abundant and widely distributed in hemi-pituitaries from the low sodium medium. Secretory granule release profiles and dense bodies were also more frequent, but the percentage of the cytoplasmic volume occupied by secretory granules was lower than on the high sodium medium. RER was only slightly modified. Significant differences were noted in the shape and processes of the non-granulated (stellate) cells of the RPD, but there were only slight differences in the ultrastructure of the somatotropes.