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Dermatofibrosarcoma protuberans treated with Mohs surgery. A case with CD34 immunostaining variability.

BACKGROUND: Immunohistochemical stains for CD34 antigen can help in differentiating dermatofibrosarcoma protuberans (DFSP) from other fibrohistiocytic tumors and in demarcating its surgical margins during Mohs surgery. However, variable expression of CD34 antigen in nodular areas can sometimes result in negative staining of tissue sections creating diagnostic and therapeutic dilemmas. OBJECTIVE: To show the variable expression of CD34 antigen in a DFSP tumor. METHODS: Case presentation of DFSP with immunohistochemical staining using the ABC peroxidase macromolecular complex. RESULTS: Microscopic examination of paraffin-embedded sections showed CD34-negative DFSP tumor cells with positive staining of endothelial cells. On frozen sections at the time of surgery the tumor cells stained strongly positive for CD34. CONCLUSION: This case demonstrates the variable expression of CD34 antigen by DFSP tumors and emphasizes the need to interpret with caution the results of these immunostains during Mohs surgery.

Adult↗

Gardnerella vaginalis has a gram-positive cell-wall ultrastructure and lacks classical cell-wall lipopolysaccharide.

Gardnerella vaginalis has a very thin cell wall with a characteristic gram-negative staining pattern and an apparent lamellar structure when viewed at an oblique angle by electronmicroscopy. Examination at right angles to the cell-wall plane and by freeze-etching showed absence of an outer membrane or any other lamellar structure. Cell-wall extracts made by methods specific for lipopolysaccharide (LPS) gave negative reactions by silver staining and for endotoxin in the limulus amoebocyte lysate assay. 2-Keto-3-deoxy-D-manno-2-octonoic acid (KDO), heptose and hydroxy fatty acids specific for LPS were not detected in the extracts. G. vaginalis cell walls are unequivocally gram-positive in their ultrastructural characteristics and chemical composition.

Cell Wall↗

Immunohistochemical study of carcinoembryonic antigen (CEA) in gastric tumors: correlation with preoperative serum levels, histologic type, and grade of anaplasia of the tumor.

The occurrence of carcinoembryonic antigen (CEA) was studied in 45 cases of gastric tumors by the immunoperoxidase technique. CEA-positive staining was found in 48.8% of tumors. A correlation was found between preoperative CEA values and tumor CEA staining. All patients with serum CEA values below 2.5 ng/ml showed CEA-negative staining of tumor. In patients with serum CEA values between 2.6 and 10 ng/ml, the tumors showed a minority of CEA-positive cells; but in patients with serum CEA values above 10 ng/ml, the tumors contained a majority of CEA-positive cells. CEA-positive staining was found in 34.4% of tumors of the diffuse type, and in 75% of tumors of the intestinal type. A high percentage of CEA positivity was seen in well-differentiated tumors (87.7%) compared to the moderately differentiated (69.2%), and to the undifferentiated (28.7%). A faint CEA-positivity was observed in intestinal metaplasia, while normal gastric mucosa was CEA-negative.

Adult↗

Ultrastructural and electron-immunocytochemical characterization of cells in epiretinal membranes.

Electron-immunocytochemical staining for three intermediate filament (IF) proteins, keratin (K), glial fibrillary acidic protein (GFAP), and vimentin (V), and for the macrophage marker, EBM/11 (E), was performed on epiretinal membranes obtained during vitrectomies performed for proliferative vitreoretinopathy (PVR), postdetachment macular puckers (PDMPs), idiopathic macular puckers (IMPs), or macular puckers associated with other disease processes. The ultrastructural and immunocytochemical characteristics of the cells were compared. Unstained cells outnumbered stained cells for each of the markers in almost all membranes. Six cell types, based on ultrastructure, were found in the majority of epiretinal membranes: 1) polarized cells with microvilli on the free border and foot processes anchoring them to extracellular matrix that consistently stained negative for all of the immunocytochemical markers; 2) spindle-shaped fibroblastlike cells that were generally negative for all markers, but rarely positive for V; 3) large undifferentiated cells with large, lightly stained nuclei and little cytoplasm that frequently expressed one of the intermediate filament (IF) proteins; 4) poorly differentiated cells that contained numerous mitochondria and frequently expressed one of the IF proteins; 5) undifferentiated, pigment-laden cells that rarely stained for any of the above IF proteins, but occasionally showed K or V positivity in a portion of the cell, suggesting that they may be losing or acquiring these proteins, and that rarely expressed GFAP; and 6) small, round, mononuclear cells with short processes that were sometimes, but not always, positive for E and that were consistently negative for K, V, and GFAP. In addition to these morphologic types, transitional cells demonstrating features of two or more of the above cell types were seen, suggesting that phenotypic changes between the various cell types can occur. The amount of extracellular matrix in epiretinal membranes showed a correlation with disease process (PVR greater than PDMP greater than IMP), and a negative correlation with the percentage of cells expressing a highly differentiated polarized morphology and with the percentage of cells staining for IF proteins. These data suggest that both cell morphology and IF protein expression may be dependent in part on microenvironment and that neither alone can be used to identify unequivocally the derivation of particular cells found in epiretinal membranes. The integration of ultrastructural and immunocytochemical data may provide a more accurate determination of the cell of origin and of phenotypic changes that have occurred. In some cases, however, both ultrastructural and IF protein composition taken together are insufficient for the precise identification of all cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Immunostaining of involucrin in odontogenic epithelial tumors and cysts.

An immunoperoxidase method was used to detect involucrin in 47 odontogenic tumors and 35 radicular cysts. Of a total of 40 ameloblastomas, 9 cases were positive for involucrin expression and those positive cases exhibited acanthomatous or follicular patterns. Squamous odontogenic tumors were strongly positive for involucrin, whereas adenomatoid odontogenic tumors gave a negative staining reaction. Involucrin expression in odontogenic tumors was divided into three categories: single cell positive, focally positive, and squamous metaplastic cell positive. Radicular cysts showed a very irregular distribution of involucrin; nonstratified epithelium was generally negative or showed only trace staining for involucrin, whereas suprabasilar stratified squamous epithelial cells were strongly positive. Cells positive for involucrin in odontogenic tumors and in cystic epithelium are probably direct signs of epithelial differentiation; such cells were squamoid in appearance.

Cell Differentiation↗

Differentiation margins of ovarian tumor pathology: first incidences of epithelial ovarian tumors monitored by marker antibodies.

The first incidence of ovarian tumors in The Netherlands was analyzed during the PALGA data. The first incidences of benign epithelial ovarian tumors reach a plateau, at a level of 60 to 65 cases per 100,000 women beyond the age of 40 years. The borderline malignant epithelial ovarian tumors account for 10 per 100,000 women aged 30 to 85, while the ovarian carcinomas reach a plateau level of 25 to 35 per 100,000 women after the age of 50. Despite the long lag period (+/- 10 years) between benign and malignant ovarian tumors, the question of whether or not all epithelial ovarian cancers develop via an intermediate step of cystadenomas is still unanswered. Therefore, we examined whether the expression pattern of intermediate filaments and tumor antigens in normal, benign, and malignant ovarian tissues might contribute to this question. The following changes in expression pattern were observed: generally, all tumor cells retained the keratin profile of the corresponding original cell type. However, in a limited number of tumor samples ectopic keratin types, such as nos. 4, 10, 13, and 14, became expressed additionally. Most epithelial ovarian tumors and mesothelial cells coexpressed vimentin. The panepithelial marker BW495/36 clearly distinguished between negatively stained normal ovarian surface mesothelium and the positively stained (inclusion) cystic epithelium. TAG-72 as well as OV-632 marked a subsequent tumor stage by discriminating between negative serous adenomas and positive serious carcinomas. TAG-72, however, stained both mucinous adenomas and carcinomas. The ovarian tumor markers (OC125, OV-TL 33, OV-TL16, MOv18), all showed an increasing expression level in the sequence order from normal cells to benign and malignant ovarian tumors. Both our epidemiological and our immunohistochemical data have shown that in the Dutch population there is a lag period of at least 10 years between the plateau levels of benign and malignant ovarian tumors. The early transformation of mesothelial cells to benign and/or malignant tumors is clearly marked by a switch on of the BW495/36 marker. Although no general transformation or progression marker from adenomas to carcinomas emerged from this study, TAG-72 might be considered a (progression) marker between the subgroup of benign and malignant serous ovarian tumors.

Age Factors↗

Expression of HLA-DR in intrahepatic cholangiocarcinoma.

Expression of HLA-DR was investigated immunohistochemically in 20 cases of intrahepatic cholangiocarcinoma, as well as 9 normal livers that had been obtained at autopsy. Five of the nine normal livers had peribiliary glands that showed HLA-DR. Positive staining for HLA-DR on tumor cells was observed in 6 of 13 cases of cholangiocarcinoma of the hepatic hilus and in only 1 of 7 cases of peripheral cholangiocellular carcinoma. In cholangiocarcinomas of the hepatic hilus, the 5-year survival rate for tumors that had positive staining for HLA-DR was better than that for tumors that had negative staining. Based on these results, the presence or absence of HLA-DR on tumor cells could have a pathologic significance for intrahepatic cholangiocarcinoma.

Adenoma, Bile Duct↗

Gastric submucosal tumours of neurogenic origin with neuroaxonal and Schwann cell elements.

Twelve cases of gastric submucosal tumours, originally diagnosed as leiomyoma and leiomyosarcoma, were investigated by staining with the neurogenic markers S-100 and neuron specific enolase (NSE). Three cases were, in addition, studied by electron microscopy. The tumours stained negatively for desmin, indicating that they were not of smooth muscle origin. All stained positively for S-100 and NSE. The ultrastructural features in the cases examined by electron microscopy were reminiscent of autonomic nerve structures normally present in the gastric wall. These included fine cytoplasmic processes of Schwann cells, wrapped with complete or incomplete basement membranes, and structures analogous to post-ganglionic neuroaxonal components. These tumours appear to represent a distinct type of gastric neoplasm originating from autonomic nerve elements in the stomach wall. They are different from previously described schwannomas or neurofibromas. In some of the tumours, identifiably neural elements are relatively a minor component, and the majority of the tumour cells are of undetermined origin. It is suggested that these cells may be poorly differentiated, lacking their antigenic determinants specific to neural differentiation.

Adult↗

Ischemia and reperfusion impair the gene expression of endogenous basic fibroblast growth factor (bFGF) in rat skeletal muscles.

Our previous studies showed that the amount of endogenous basic fibroblast growth factor (bFGF) was reduced after ischemia and reperfusion insult. One of the mechanisms involved in the decrease of endogenous bFGF is the increased destruction of this growth factor associated with oxygen free radical activation and inflammation. We hypothesized that the wounding also impairs the secretion of bFGF and examined the bFGF gene expression in skeletal muscles after ischemia and reperfusion insult. In this study, a rat leg ischemia (4 h) and reperfusion (24 h) injury model was prepared and the in situ hybridization method and reverse transcriptase polymerase chain reaction technique (RT-PCR) were used to evaluate the bFGF gene expression and its localization in control (normal) and injured rat skeletal muscles. The results showed that the bFGF mRNA expression was localized in the cytoplasm in control skeletal muscle, especially at the periphery inside the cells. According to the intensity of the stain, four main classes of fibers could be identified: strongly, moderately, weakly, and negatively stained fibers. Based on the positive stain, about 82% of the total fibers examined were positive for bFGF mRNA stain. In ischemic or ischemic and reperfused rat skeletal muscles, the localization of bFGF mRNA expression was similar to that in normal skeletal muscles, but only 52% in ischemic muscles and 22% in ischemic and reperfused muscles had positive bFGF mRNA staining. RT-PCR confirmed a significant decrease in bFGF mRNA expression in ischemic and reperfused rat skeletal muscles. These results suggest that the acute ischemia and reperfusion not only induce the destruction of endogenous bFGF molecule, which is stored at the extracellular matrix of the fibers, but also downregulate the bFGF gene expression. The simultaneous dysregulation of endogenous bFGF gene expression and decreased synthesis of bFGF protein suggest a possible role of this growth factor in delayed wound healing.

Animals↗

A myristylated membrane protein encoded by the vaccinia virus L1R open reading frame is the target of potent neutralizing monoclonal antibodies.

We identified a protein component of the intracellular mature vaccinia virion membrane that is a target of a potent neutralizing monoclonal antibody, 7D11, obtained from Alan L Schmaljohn. By immunofluorescent and electron microscopic analysis, MAb 7D11 was found to stain intracytoplasmic viral factories, virion membranes in cell sections, and the surface of negatively stained preparations of purified virions. The MAb 7D11 antigen, which is synthesized at late times in infection, has apparent molecular masses of 25 and 29 kDa under nonreducing and reducing conditions, respectively. The membrane antigen was most efficiently extracted from virions by NP40 detergent in combination with a reducing agent; in addition, the protein partitioned exclusively into the detergent phase when extracted with Triton X-114. Although the N-terminus of the immunoaffinity-purified protein was blocked, sequence analysis of trypic peptides revealed that the MAb 7D11 antigen was identical to the myristylated protein encoded by the L1R open reading frame previously described by C.A. Franke, E.M. Wilson, and D.E. Hruby (1990, J. Virol. 64, 5988-5996). Validation of this genetic assignment was provided by the ability of MAb 7D11 to immunoprecipitate a [3H]myristic acid-labeled product of the expected molecular weight from infected cells. In addition, we discovered that the previously described neutralizing monoclonal antibody 2D5 (Y. Ichihashi, T. Takahashi, and M. Oie, 1994, Virology 202, 834-843) also recognizes the L1R protein.

Antibodies, Monoclonal↗

The immunocytochemical localization of GFA protein in experimental murine CNS tumors.

Immunocytochemical localization of GFA protein in formalin-fixed, paraffin-embedded tissue sections by the peroxidase-antiperoxidase method of Sternberger was used to study experimental murine CNS tumors. Transplacental tumor induction in rats by ethylnitrosourea produced oligodendrogliomas and mixed gliomas in the cerebrum and spinal cord, and malignant Schwannomas of the trigeminal nerve. A methylcholanthrene-induced mouse "ependymoblastoma" inoculated intracerebrally in normal and in toxoplasma-infected mice was also studied. A positive reaction of GFA protein antibody was seen in the astrocytic portion of the mixed gliomas; the oligodendrogliomas, the malignant Schwannomas and the mouse "ependymoblastoma" were negative. Staining for GFA protein delineated the astrocytic reaction of neural tissue adjacent to the tumors. The reaction was markedly intensified in the brains of mice infected with toxoplasma. Additionally, ependymal cells near the tumors stained positively for GFA protein; normal ependyma at a distance from tumor remained negative. The technique, which combines a high degree of specificity with great sensitivity and is readily adaptable to routinely processed tissue, should prove a valuable tool in experimental oncology of the central nervous system.

Animals↗

Morphological changes in HAV-infected Frp/3 cells and immunolocalization of HAAg.

Electron and immunoelectron microscopic studies were carried out on HAV-infected Frp/3 cells. The infection led to a distinctive cytopathic effect (CPE) arising on day 3 up to the complete detachment of monolayers on day 7. Infected cells exhibited progressive modifications, beginning from the formation of long helical polyribosomes. Subsequently, hypertrophy, cisternal dilatation and degranulation of the RER could be observed. Furthermore, the formation of concentric membranous bodies (CMB), large myelin-like structures and annulate lamellae could be revealed at later times of infection. 24-27 nm virus-like particles were observed within cytoplasmic vesicles or outside extensively degenerated cells. Indirect immunoperoxidase staining were used to localize HAV antigen (HAAg) in thin sectioned infected Frp/3 cells. Vesicular inclusion bodies, often seen to contain electron-lucent particles, resulted darkly stained as well as tracts of the RER and myelin-like structures. Negatively stained preparations from cell lysates revealed small clusters of HAV particles which sometimes appeared to be still associated with residual membrane fragments. Our findings seem to suggest that HAV replication occurs in close association with cytoplasmic membranes and a direct involvement of the RER seems to be demonstrated.

Antigens, Viral↗

Granular acute lymphoblastic leukemia with hypereosinophilic syndrome.

A four-year-old boy presented with marked peripheral blood eosinophilia (absolute eosinophil count of 54 x 10(9)/1), features of hypereosinophilic syndrome, and acute lymphoblastic leukemia (ALL-L2), the latter characterized by the presence of granular blasts. Blasts were negative for myeloperoxidase, non-specific esterase, acid phosphatase, periodic-acid Schiff stain, and toluidine blue. They exhibited an early pre-B immunophenotype (TdT, CD19, CD10, CD20 and CD22 positive) and stained negative for T (CD7, CD2, CD5 and CD3) and myeloid markers (MPO, CD33 and CD13). Chromosomal analysis revealed a normal karyotype. To the best of our knowledge, this case represents the first report of the coexistence of granular ALL and hypereosinophilic syndrome.

Child, Preschool↗

Multifocal micronodular pneumocyte hyperplasia in a postmenopausal woman with tuberous sclerosis.

We report a peculiar case of multifocal micronodular pneumocyte hyperplasia (MMPH) without association of pulmonary lymphangioleiomyomatosis (LAM) in a 56-year-old postmenopausal woman with tuberous sclerosis. This case is surmised to be a forme fruste of tuberous sclerosis. Computed tomography demonstrated multiple micronodules, measuring up to 5 mm in size, present in the bilateral lung fields, but no cystic changes. A proliferation of pleomorphic type-II pneumocytes lining the thickened alveolar septa in an adenomatoid pattern, with an associated increase in alveolar macrophages, was observed without typical nuclear atypia. In fully developed lesions, the ingrowth of more proliferating type-II pneumocytes into the thickened alveolar septa and macrophages filling the alveolar lumens were characteristic findings. Proliferation of immature smooth muscle cells suggesting LAM was not observed. Positive immunohistochemical stains for cytokeratin, epithelial membrane antigen, and surfactant apoproteins A and B, and negative staining for HMB45, alpha-1 smooth muscle actin, desmin, and carcinoembryonic antigen confirmed the characteristics of alveolar lining cells in each MMPH lesion. MMPH associated with tuberous sclerosis in the postmenopausal woman appears to be similar to that described in premenopausal women. The present case is familial rather than sporadic and suggests no relationship between the development of MMPH and the underlying hormonal state.

Female↗

Thyroglobulin in carcinoma of the thyroid: an immunohistochemical study.

To determine the value of thyroglobulin as an immunohistochemical marker for thyroid neoplasms, we studied 42 primary thyroid carcinomas, 38 metastatic carcinomas, and four sarcomas involving the thyroid gland. All follicular and papillary carcinomas, regardless of their morphologic variation, stained positively for thyroglobulin, whereas the medullary carcinomas, metastatic tumors, and sarcomas showed negative staining reactions. The only small-cell variant of follicular carcinoma and ten of 14 spindle and giant cell carcinomas showed the lowest thyroglobulin reactivity. It is concluded that immunohistochemical demonstration of thyroglobulin is a sensitive and specific method of identifying thyroid carcinomas of follicular cell origin.

Carcinoma↗

Benign fibrous histiocytoma of the skin. An immunohistochemical analysis of 30 cases.

In this study the immunohistochemical analysis of distinct morphologic variants of benign fibrous histiocytoma (BFH) of the skin was performed with immunoperoxidase technique for both lysozyme and alpha-1-antitrypsin (A1AT). Thirty cases including cellular, fibrous and xanthomatous variants of BFH were selected. Out of the total 6 cases (20%) showed positive staining only for A1AT, 3 cases (10%) only for lysozyme and 10 (33.3%) for both markers. Thus, 19 cases (63.3%) showed positive staining for one of both markers. Positive staining was higher in the cellular variant than the fibrous and xanthomatous types. Negative staining of tumors of definite histiocytic morphology may be interpreted as a variable enzymatic expression of different histiocytic activation and/or undetectable enzymatic content by the current techniques. These results are in accordance with our previous evolutional hypothesis of BFH which considered the cellular tumors as functionally more active variants evolving to less cellular, more fibrous and less active types. Current histogenetic concepts about this controversial group of skin neoplasms are discussed.

Aged↗

Distribution of type I and type III collagen in the developing periodontal ligament of mice.

In order to localize type I and type III collagen in developing periodontal ligament, immunofluorescent and immunoelectron microscopic examinations were undertaken. The materials used were the maxillary molars of CF1 mice, 13 days to 6 months old. The antibodies used were monospecific polyclonal antibodies against type I or type III collagen raised in rabbits or guinea pigs, respectively. Single- and double-staining methods were employed in immunofluorescent as well as immunoelectron microscopic examinations. In immunofluorescent examinations, during the development of the periodontal ligament, periodontal fibers showed intense and homogenous staining for both type I and type III collagen; while Sharpey's fibers in the alveolar bone showed a heterogenous staining in cross and longitudinal sections for both types of collagen. In immunoelectron microscopic examinations, fibrils of periodontal fibers showed positive staining for type I and type III collagen simultaneously and had characteristic cross-banding and had a large diameter (ranging 40 approximately 80 nm) which remained constant during development. Sharpey's fibers in the alveolar bone or in the cementum showed the same staining pattern as the periodontal fibers, except for an afibrillar area which showed negative staining for both type I and type III collagen. The results obtained suggest that periodontal fibers and Sharpey's fibers consist of cofibrils of at least type I and type III collagen.

Aging↗

[Overexpression of c-erbB-2 oncogene in primary ovarian cancers: incidence and prognostic significance in 243 patients].

In ovarian cancers, no valuable prognostic factors are available so far for predicting biological behaviour and clinical outcome. During the last years, overexpression of c-erbB-2-oncogene has been reported to be a prognostic factor in various human carcinomas. Its prognostic value in ovarian cancer is still a matter of controversy. The records and histological specimens of 243 patients with primary ovarian cancer were reviewed. Overexpression of c-erbB-2-oncogene encoded transmembrane protein p185 was determined immunohistochemically using polyclonal and monoclonal antibodies. Of 243 investigated tumours, 45 (19%) were positive. We examined the relationship between p185 and clinical stage, histological type, grading and prognosis. p185 overexpression differs within the histological subtypes of tumours. No correlation was found between p185 overexpression and stage or histological grade. The follow-up data demonstrate that patients with positive staining for p185 have a significantly worse prognosis (p = 0.001) than those with negative staining. These results suggest that overexpression of c-erbB-2 oncogene is associated with higher biological aggressiveness and unfavourable course of disease.

Adenocarcinoma↗