Macroscopic, histologic, histochemical, immunohistochemical, and ultrastructural features of mesothelioma

Mesotheliomas are uncommon neoplasms that arise from the cells forming the serosal membranes of the body cavities. Approximately 90-95% of mesotheliomas arise in the pleural cavity and 5-10% in the peritoneal cavity. Rare mesotheliomas arise in the pericardium and in the tunica vaginalis. Unlike many neoplasms, mesotheliomas grow in a diffuse distribution and tend to encase the organs in the various body cavities. A combination of histochemical, immunohistochemical, and ultrastructural features are often necessary to accurately diagnose mesotheliomas. These techniques are highlighted in this review article on mesothelioma.     

Granulated lymphocytes in human endometrium: histochemical and immunohistochemical studies.

Granulated lymphocytes with an unusual antigenic phenotype (CD56+ CD38+ CD2 +/- CD3- CD16-) form a substantial proportion of leukocytes in human endometrial stroma. The purpose of this study was to examine morphological and antigenic heterogeneity in endometrial granulated lymphocytes (eGL) in imprint preparations, paraffin-embedded sections and frozen sections. eGL in decidual imprints showed variations in cell size, nuclear size, shape and chromatin content and the number and size of cytoplasmic granules. eGL were detected in paraffin-embedded sections using phloxine tartrazine, alcian blue and toluidine blue stains. There was no difference in the number of eGL among the three stains but the granules appeared smaller and more regular when stained with toluidine blue. The proportion of stromal cells which were leukocytes increased from 8.2% in proliferative endometrium to 31.7% in early pregnancy decidua. The number of CD56+ and CD38+ cells increased in late secretory endometrium; CD56+ cells formed greater than 75% of the leukocytes in first trimester decidua. The increased number of CD2+ cells in decidua was not comparable with CD56+ and CD38+ cells suggesting that a lower proportion of CD56+ cells in first trimester decidua coexpress CD2, an observation which was supported by double labelling studies. eGL therefore show morphological and antigenic heterogeneity and the study of granulated lymphocytes in pathological endometrium and decidua will require careful phenotypic and morphological analysis of accurately dated samples.     

An immunohistochemical study of Warthin-Finkeldey cells in measles

An autopsy case of an 18 month old male infant with measles infection Is reported. An autopsy revealed generalized lym-phadenopathy, splenomegaly and hyperplastic thymus. Histologically, there were many Warthin-Finkeldey cells (WFC) in hyperplastic lymphoid tissues. Although viral particles and inclusions were not detected electronmicroscopicaliy, the infection of measles virus was detected in WFC by immunofluorescent technique using anti-measles antibody. Warthin-Finkeldey cells were morphologically and immu-nohistochemically divided into two groups. The first type was WFC appearing in the germinal centers of lymphoid tissue, revealing many large nuclei and B cell markers. The second type was WFC appearing in the interfollicular areas and thymus, showing many small hyperchromatlc nuclei and T cell markers. The data indicate that there might be a morphologic and immunophenotypic heterogeneity in WFC.     

Low-grade tubular-mucinous renal neoplasm with neuroendocrine differentiation: a histological, immunohistochemical and ultrastructural study

Low-grade tubular-mucinous renal neoplasm (LGTMRN) was recently established as a distinct carcinoma classification. A 70-year-old, female traffic accident victim underwent a detailed examination that disclosed a huge mass in the lower pole of the left kidney. The patient underwent a nephrectomy based on a diagnosis of renal tumor. Macroscopically, the tumor was well demarcated and a whitish color with focal hemorrhage. Histological examination showed that tumor cells proliferated through tubular, trabecular, and solid growth patterns in the mucinous background. Focally, foci of clear cells or the proliferation of spindle cells was also observed. Nuclei were generally round and uniform in size. No abnormal mitotic figures were identified. Immunohistochemically, tumor cells were diffusely positive for AE1/AE3, vimentin and chromogranin A, and focally positive for cytokeratin (CK) 18, CK19, Ulex europaeus agglutinin-1, epithelial membrane antigen, neuron-specific enolase (NSE), CD9 and CD57. Ultrastructurally, tumor cells contained a moderate number of mitochondria, rough endoplasmic reticulum and dense-core granules. No renin granules or glycogen were observed. Microvilli were focally seen. Our results render further evidence that LGTMRN is a distinct entity from the hitherto established renal neoplasms. Foci of clear cells and neuroendocrine differentiation should be added to the histological spectrum of LGTMRN.     

Histological and histochemical analysis of embryoid bodies

We examined the histological structure of embryoid bodies arising from aggregation of mouse embryonic stem (ES D3) cells after 7, 12, 18 and 26 days of in vitro culture. Morphology of originally solid embryoid bodies was affected by the process of cavitation that resulted in formation of cystic embryoid bodies and by spontaneous differentiation of the ES D3 cells. We applied in situ immunophenotyping to characterise cell populations that spontaneously differentiated inside the embryoid bodies in the various stages. The most distinct cell populations that were found inside embryoid bodies were alpha-fetoprotein-positive endodermal cells and myogenic cells that expressed desmin, myogenin or smooth muscle actin. ES D3-derived endothelial cells generated during vasculogenesis inside the embryoid bodies differed from mature endothelial cells because they did not stain for von Willebrand factor. These cells also differed from endothelial cell that were generated during angiogenesis since they did not stain for the intermediate filament nestin. Our results demonstrate the usefulness of this in vitro model for studying early embryogenesis.     

Chin-marking behavior in male and female New Zealand rabbits: onset, development, and activation by steroids.

Chin marking (chinning) frequency was determined daily in 25 male and 24 female New Zealand rabbits aged 31-150 days. Chinning appeared earlier in females (mean +/- SD = 41 +/- 16 days) than in males (47 +/- 13 days). Between days 30-50, females displayed chinning more frequently than males. Thereafter, chinning increased steadily in both sexes up to day 100. Chinning curves levelled on days 100 and 140 in males and females, respectively. Profile analysis of male and female chinning curves showed significant differences in their slopes and in their population means (p < 0.001). At 7 months of age (days 210-224), both sexes displayed adult chinning frequencies: mean of means +/- SD = 103 +/- 18 and 79 +/- 14 marks/10 min in males (n = 8) and females (n = 8), respectively. The administration of testosterone propionate (TP, 1 mg/day) or estradiol benzoate (EB, 1 microgram/day) to males and females, respectively, from days 31-50, stimulated higher chinning frequencies than those displayed by untreated animals. Results suggest that chinning frequency increases throughout development largely as a consequence of a concomitant increase in sex steroid secretion.     

Progestins used in hormonal replacement therapy display different effects in coronary arteries from New Zealand white rabbits

Objectives: The aim of this study was in an animal model to assess the vascular effects of different progestins commonly used in hormonal replacement treatment. Methods: Fifty-six non-atherosclerotic, ovariectomized New Zealand white rabbits were randomized into seven groups: (1) medroxyprogesterone acetate (MPA), (2) norethisterone acetate (NETA), (3) conjugated equine estrogens (CEE), (4) 17-β-estradiol (E2), (5) MPA+CEE, (6) NETA+E2, (7) or placebo (n=8) and given hormonal treatment through the diet for 4 weeks. Ring segments from the left proximal coronary artery and from the distal part of the left anterior descending coronary artery were microdissected and mounted for isometric tension recordings in a myograph. The vasoconstrictory responses induced by potassium, endothelin-1, calcium and N_w-nitro- -arginine methyl ester, and the vasodilatory response induced by acetylcholine and sodiumnitroprusside were investigated. The maximum contraction/relaxation (E_max) and the concentration required to induce half the maximum response (EC₅₀) were determined. EC₅₀ values were expressed as the negative logarithm to the molar concentration, pD₂=−log EC₅₀. Results: Treatment with MPA alone caused when compared to treatment with NETA an increase in tension development in the distal coronary artery after the addition of potassium (6.36±0.36 versus 4.31±0.42 P<0.005) (single dose response, mN/mm, mean±S.E.M.) and endothelin-1 (9.41±0.82 versus 6.43±0.73 P<0.05) (E_max, mN/mm, mean±S.E.M.). Treatment with MPA compared to placebo caused an endothelin-1 induced increase of E_max in the distal coronary artery (9.21±0.87 versus 6.51± 0.65 P<0.05) and a calcium induced increase of pD₂ in both coronary arteries (2.98±0.19 versus 2.42±0.12 P<0.05, proximal coronary artery) (3.26±0.09 versus 2.9±0.1 P<0.05, distal coronary artery) (pD₂, mean±S.E.M.). Treatment with NETA compared to placebo in the proximal coronary artery, after the addition of sodiumnitroprusside caused a decrease of pD₂ (5.33±0.19 versus 5.94±0.13 P<0.05). Treatment with E2 compared to treatment with CEE in the proximal coronary artery caused a decrease of pD₂ after the addition of sodiumnitroprusside (5.00±0.16 versus 5.77±0.28 P<0.05). No significant differences were found between MPA+CEE and NETA+E2. Conclusion: Treatment with MPA alone seems to enhance the contractile response to potassium and endothelin-1 in the distal coronary artery compared to NETA, indicating that different progestins used in hormonal replacement treatment may display different effects on contractile functions of coronary arteries.     

Development of the notochord in human embryos: ultrastructural, histochemical, and immunohistochemical studies

In the present study of the notochord, the specimens were 54 externally normal human embryos ranging between Carnegie stages 13 and 23. The following staining procedures were used: periodic acid-Schiff (PAS), modified method of PAS, alcian blue, colloidal iron, and toluidine blue. Routine electron microscopic techniques were used. Immunoreactivity of the notochord to alpha-enolase was also examined. The notochord cells were undifferentiated in stage 13 with few intracellular organelles. The microfibrils and deposition of acid mucopolysaccharides appeared in the notochordal sheath in stage 14. The characteristic relation of mitochondria with rough endoplasmic reticulum was observed. Golgi complexes increased in the perinuclear region in stage 15. The layer of microfibrils in the notochordal sheath initially separated from the notochord in stage 16. Glycogen, mucoprotein, neutral mucopolysaccharides, and glycolipids began to increase in the mesenchymal cells around the notochord, starting at stage 16. Acid mucopolysaccharides increased in the notochordal sheath and in the matrix of the precartilage area around the notochord as this embryonic stage advanced. It was also revealed that the immunoreactivity of the notochord to alpha-enolase remained constant during the embryonic period. The results show that the notochord is transformed from an apparently undifferentiated organ into an organ with secretory activity in stage 14, producing microfibrils and depositing acid mucoplysaccharides in the notochordal sheath. The immunoreactivity of the notochord to alpha- and gamma-enolase isoenzymes and the development of the notochord are discussed. This study was undertaken to provide additional information on the development of tumors of notochordal origin.     

Adverse effects of the anabolic steroid, boldenone undecylenate, on reproductive functions of male rabbits

This study was conducted to evaluate the adverse effects of the anabolic steroid, boldenone undecylenate (BOL) on reproductive functions of male rabbits. Thirty white New Zealand mature male rabbits were divided into three groups (10 rabbits each). Group A rabbits served as a control group. Group B rabbits received 4.4 mg/kg body weight (bwt) BOL 5% oily solution. Group C rabbits received 8.8 mg/kg bwt BOL. Rabbits were injected intramuscularly twice weekly for two months. BOL had no significant effect on the bwt and bwt gain. Testes and epididymis weights were decreased significantly in the BOL-treated groups. BOL caused significant reduction in serum testosterone level, seminal volume, sperm motility, and sperm count. No abnormalities were detected in the sperm morphology of the BOL-treated groups. Histopathological alterations in the testes and epididymis were marked in the group C rabbits. These results indicate that administration of BOL exerts a significant harmful effect on the reproductive functions of male rabbits.     

Differentiation between ulcerative colitis and Crohn's disease by a quantitative immunohistochemical evaluation of T lymphocytes,neutrophils, histiocytes and mast cells

Mucosal biopsy criteria has limited validity in terms of discrimination between ulcerative colitis (UC) and Crohn's disease (CD). The aim of this study was to set up quantitative immunohistochemical criteria, with a special focus on inflammatory cell distribution within individual specimens and throughout the large bowel. Quantitative evaluation was performed for the density of CD8+, CD45RO+, neutrophil elastase+, CD68+ and mast cell tryptase+ cells in affected and unaffected mucosa taken from 41 patients with UC and 61 patients with CD. Each slide was examined at the highest and lowest density fields, which were further divided into the upper and deeper half of mucosa. Multiple logistic regression analysis using 51 features as independent variables constructed a predictive equation finding the probability of UC (PUC), and the diagnostic categories were subsequently defined based on a receiver-operating characteristic curve. The analysis disclosed five significant features suggesting UC; these implied intense infiltration of CD8+ and mast cell tryptase+ cells, diffuse infiltration of neutrophil elastase+ and CD68+ cells, and continuous infiltration of CD45RO+ cells. The criteria consisted of three diagnostic categories, 'suggestive of UC (PUC > or = 0.7)', 'indeterminate (0.3 < PUC < 0.7)', and 'suggestive of CD (PUC < or = 0.3)'; the criteria had values for sensitivity and specificity exceeding 95%. The immunohistochemical criteria distinguishing UC from CD may help to confirm the diagnosis in patients with ambiguous endoscopic and histological diagnosis.     

Metanephric adenoma of the kidney: Ultrastructural, immunohistochemical and lectin histochemical studies

A case of metanephric adenoma, a rare benign tumor of the kidney is reported, and the results of ultrastructural, immunohistochemical, and lectin histochemical studies are presented. The patient was a 47 year old Japanese man presenting wlth a left renal tumor by ultrasonography. The nephrectomy material revealed a well-demarcated tumor with medullary appearance, and on histological examination the tumor was composed of cuboidal cells forming regular small tubules with hyallnous stroma. Incomplete glomeruloid structures were formed in a few elongated tubules com-posed of columnar cells. Metanephric blastema was not observed in the tumor or in the normal kidney. By electron microscope, the tumor cells were observed to be immature cells with microvilli at the apical surfaces. The neoplastic tubules were surrounded by basal lamina. The tumor cells were immunoreactive for Leu 7, epithelial membrane antigen, vimentin, and low molecular weight cytokeratin, and were weakly immunoreactive for S-100 protein. Peanut agglutinin, soybean agglutinin, and Dollchos bifiorus agglu-tinin were bound to the apical surfaces of the tumor cells.     

Signet-ring stromal tumor of the ovary: A histochemical,immunohistochemical and ultrastructural study

We report a signet-ring stromal tumour found in the ovary of a 50-year-old woman. The solid tumour was composed of spindle and round cells which stained with vimentin and some round cells were vacuolated and had the appearance of signet-ring cells. The vacuoles were negative with periodic acid-Schiff, with and without diastase digestion, with alcian blue, Sudan III, alpha-1-antitrypsin and alpha-fetoprotein stains. Ultrastructurally, they were membrane bound and contained scanty flocculent material. Some neoplastic cells contained intracytoplasmic eosinophilic globules which were positive with alpha-1-antitrypsin. Signet-ring stromal tumour is a benign, morphologically distinctive type of ovarian stromal tumour, which may present problems in differential diagnosis with other stromal and non stromal ovarian neoplasms.     

Histiocytoid breast carcinoma: Histological, immunohistochemical, ultrastructural, cytological and clinicopathological studies

Histiocytoid breast carcinoma (HBC) is a rare variant of breast carcinoma and often causes a diagnostic dilemma because of its histological similarities to some types of breast cancer and benign lesions. To elucidate the incidence of HBC and its biological properties, histological specimens from 1010 breast cancer patients treated at Yokohama Minami Kyosai Hospital between 1972 and 1996 were reviewed. Three cases of pure HBC and three cases of combined HBC (two with pleomorphlc lobular carcinoma and one with apocrine ductal carcinoma) were found, yielding an Incidence of 0.3% for each. Two of the three pure HBC cases contained foci of in situ lobular carcinoma. Targetoid and Indian file invasive patterns, the features characteristic of lobular carcinoma, were present in all three pure HBC cases and in two of the three combined HBC with pleomorphic lobular carcinoma. These results, together with those of previous studies, suggested that the majority of HBC are of lobular origin, although the apocrine ductai origin is also possible in a small number of HBC. Diastase-resistant periodic add-Schiff-positive granules and granular immunoreactivities for gross cystic disease fluid protein-15 (GCDFP-15) were characteristic of the histiocytoid tumor cells in both the pure and combined HBC, suggesting the apocrine differentiation of tumor cells. All three pure HBC cases were in stage 1 and were free of the disease for up to 5 years and 1 month after the lumpectomy. Thus, the prognosis of HBC appears to be dependent on the stage of the disease and may not always be poor, as indicated by the original report mentioning a preferential eyelid metastasis.