Search

Background: Vascular enumeration using antibodies to Factor VIII has been reported to be an independent prognostic indicator of invasive breast carcinoma., Methods: To eliminate potential subjectivity in distinguishing between individual vessels, especially in areas of tangled capillaries, total endothelial area (EA) was assessed using a Samba 4000 image analyzer. One hundred seventy-eight invasive breast carcinomas (Stage 1 and 2, mean follow-up: 71 months) were immumostained for the presence of CD34, the human hematopoietic progenitor cell antigen also present in endothelium, and EA was quantitated within 5 adjacent 20X fields (0.74 mm2). Additionally, these same vessels were manually counted from the image analyzer. Manual counts were also made from a photomicrograph representative of a single 10X field (1.06 mm2)., Results: High grade carcinomas contained greater endothelial area than low grade carcinomas (P = 0.0001). Endothelial area was prognostically significant (P = 0.004) in univariate analysis of disease-free survival (DFS) and overall survival (OS), as were stage of disease, tumor size, and combined histologic grade (P < or = 0.024). Manual vessel counts from the monitor were significant for OS only. Manual vessel counts from photomicrographs showed no statistically significant association with DFS or OS. In multivariate analysis, EA, but not vessel enumeration, remained as an independent predictor for OS (lymph node negative patients only, n = 87) and for DFS (lymph node positive patients only, n = 91). For the entire group of patients (lymph node negative and lymph node positive) independent predictors of DFS and OS were tumor grade and size (P < or = 0.006)., Conclusions: Of the three methods used to evaluate tumor angiogenesis, total endothelial area, as objectively evaluated by image analysis, was the only independent prognostic indicator for OS for patients with lymph node negative invasive breast carcinoma.

Background: Expression of carcinoembryonic antigen (CEA) has been reported in 10-95% of breast cancer. Its value as a predictor of disease progression is controversial., Methods: The expression of CEA in 202 Stages I and II breast carcinomas was assessed by immunohistochemistry, and the results were correlated with various histologic and clinical parameters to establish CEA's biologic relevance. The mean follow-up of the patients was 6.5 years. The monoclonal antibody used does not cross-react with other molecules in the CEA gene family., Results: One hundred, thirteen (56%) tumors expressed CEA in more than 15% of the cells. Expression of CEA was associated with positive estrogen receptor (ER) status (P = 0.003). Univariate Cox regression analysis showed that, whereas disease free survival (DFS) and overall survival (OS) were not associated significantly with CEA expression, tumor size, nuclear grade, ER status, lymph node metastases, and stage were. When ER status was stratified to CEA expression, patients who were ER negative and had CEA-negative tumors had a 3.9 times higher risk (P = 0.032) of death than did the patients with CEA-positive tumors. Cox regression analysis revealed that ER was the only parameter with significant interacting effect with CEA. Multivariate, stepwise Cox regression analysis showed that CEA expression, tumor size, and nuclear grade were the only significant independent predictors of DFS, and nuclear grade and lymph node metastasis the only significant predictors of OS in the ER-positive group. The only significant independent predictor of DFS and OS in the ER-negative group was CEA. When CEA expression was stratified to ER status, patients whose tumors lacked CEA and ER had threefold higher risk of disease relapse (P = 0.002) and a 5.3-fold higher risk of death (P = 0.0001) than those with ER-positive and CEA-negative tumors. Multivariate analysis showed that the association between CEA and ER was enhanced further after compensating for other parameters with independent predictive value., Conclusions: The association between CEA and ER was the most important independent predictor of a subgroup of patients (CEA-negative, ER-positive) with the most favorable prognosis. The results imply that the association of several tumor markers may provide tumor profiles with superior predictive value than a single parameter.

Background: Cathepsin D is an estrogen-regulated lysosomal protease that may be overexpressed in breast cancer. Several studies based on biochemical analysis of tumor cytosol have shown that high levels of cathepsin D are associated with poor outcome in patients with breast cancer. In contrast, a few immunohistochemical studies have shown that cathepsin D positivity in breast cancer cells indicates a favorable prognostic outcome or is of no prognostic significance., Methods: Because of the current controversy, the authors studied, using immunohistochemistry, the distribution (0 to 3) and intensity (0 to 3) of cathepsin D in Stage I and II infiltrating ductal carcinoma of the breast from 245 patients treated at the City of Hope National Medical Center, Duarte, California, from 1970 to 1986. A commercially available rabbit antiserum to purified human cathepsin D and the standard avidin-biotin complex method were used. Statistical analysis was based on a combined low (0 or 2) or high (3 to 6) score., Results: A high cathepsin D score was associated with Stage II disease (P = 0.007), positive lymph nodes (P = 0.019), high nuclear grade (P = 0.005), and c-erbB-2 oncogene amplification (P = 0.013). Cathepsin D positivity was not an independent prognostic indicator of disease-free survival (DFS) or overall survival (OS). In a subgroup analysis, the presence of cathepsin D had only a modest association with predicting a shorter DFS in patients with negative lymph nodes (P = 0.072) or positive progesterone receptors (PR) (P = 0.086)., Conclusions: Immunohistochemical analysis of cathepsin D, with the antiserum used in this study, is not an independent predictor of outcome in patients with breast cancer because of its strong associations with several well-established prognostic indicators.

Background: Squamous cell lung carcinoma (SLC), the most frequent type of lung cancer, generally is treated surgically and its prognosis is poor. The only current clinically useful prognostic criterion is lymph node staging (TNM classification). Expression of a novel tumor-associated carbohydrate epitope Gal beta 1-3[Fuc alpha 1-4]GlcNAc beta 1-4[Fuc alpha 1-3]GlcNAc beta 1-3 Gal beta 1-4Glc identified by the 43-9F monoclonal antibody (MoAb) is associated with the growth pattern of SLC cell lines in athymic mice and in vitro. This implies that the 43-9F epitope may be related to tumor progression in patients with SLC and that, as such, it could be of prognostic value., Methods: Primary tumor specimens from 231 patients with lung carcinoma (130 with SLC, 64 with adenocarcinoma, 10 with small cell carcinoma, 16 with large cell carcinoma, and 11 with adenosquamous carcinoma) were examined by immunohistochemical studies on formalin-fixed, paraffin-embedded tissue samples for immunoreactivity with an MoAb to the 43-9F antigen. Univariate and step-wise Cox regression analyses were used to compare survival time by histopathologic diagnosis, smoker status, TNM classification, and type of surgical treatment., Results and Conclusions: Patients with 43-9F epitope-positive SLC tumors had a significantly (P less than 0.01) better prognosis than patients with epitope-negative tumors. In contrast, no association was seen between 43-9F epitope expression and survival time for patients with lung adenocarcinomas. Further, the prognostic value of 43-9F expression in SLC was found to be superior to the N-classification with the added advantage that it requires access only to primary tumor tissue and thus is available before therapy.

The authors recently reported the antigenic phenotypes of three cases of so-called "malignant angioendotheliomatosis" and suggested that angiotropic large cell lymphoma (ALCL) is a more appropriate designation for this disease. The authors now report an additional seven cases of ALCL with unique clinical presentations. One patient presented with prostate enlargement, the second with lytic bone lesions and thickened nasal sinus mucosa, the third had diffuse myalgia, the fourth had dyspnea and pulmonary infiltrates, the fifth had gangrene of the lower extremities, total-body skin involvement, and pancytopenia, the sixth had a lesion of the foreskin mimicking squamous cell carcinoma, and the seventh had a mediastinal mass. In all cases histologic features were characteristic of ALCL with, in two cases, extravascular spread into soft tissue. Immunohistologic studies showed a B-cell phenotype in five cases and a T-cell phenotype in one case. Two patients received combination chemotherapy using established treatment protocol for large cell lymphoma, and remain in complete clinical remission and two patients are responding clinically to combination chemotherapy. Two patients died shortly after receiving combination chemotherapy. One patient has only recently been diagnosed as having ALCL and no long-term follow-up is available. These data indicate that, although ALCL affects predominantly the central nervous system and skin, unusual clinical presentations may occur, and patients with ALCL may respond to combination chemotherapy for large cell lymphoma.

Paraffin sections of formalin-fixed tumor samples from 26 patients with neuroendocrine (Merkel cell) carcinoma of the skin (NECS) were studied immunohistochemically with three monoclonal antibodies to low molecular weight keratin (MAB-K) and with antibodies to leukocyte common antigen (LCA), neurofilament (NF), neuron-specific enolase (NSE), S100 protein (S100), and chromogranin (CGN), to investigate the relative diagnostic value of these antibodies. Samples from 20 lymphomas, 10 non-oat cell undifferentiated carcinomas, 10 oat cell carcinomas, and 10 melanomas served as controls. Keratin was found in 25 of the 26 NECS and in all undifferentiated and oat cell carcinomas. A ball-like immunostaining for keratins, resembling an inclusion body was seen only in cases of NECS and some carcinoids. Neurofilament, NSE, and CGN were expressed by fewer NECS than was keratin and all NECS were negative for LCA and S100. None of the lymphomas and melanomas contained detectable keratin, NF, NSE, or CGN. Only the lymphomas stained with LCA. Only the melanomas were S100-positive. It is concluded that keratin is the most useful single discriminating marker in the separation of neuroendocrine (Merkel cell) carcinoma of the skin from lymphoma, melanoma and, when the characteristic inclusion-like pattern is seen, from metastatic oat cell carcinoma.

Alpha-lactalbumin, the B protein of lactose synthetase secreted by the mammary epithelial cells, was isolated and purified from fresh human milk and injected into rabbits for antibody production. Indirect immunofluorescence (IIF) was done on various types of breast lesions to assess the relationship, if any, between histologic type and production of alpha-lactalbumin. Fifty percent of fibroadenomas and fibrocystic disease and 63% of infiltrating ductal carcinoma showed positive reaction. No false positives were found on IIF of alpha-lactalbumin of several tumors of nonmammary tissue. These findings suggest that IIF for alpha-lactalbumin may be helpful in ascertaining the site of origin of metastatic breast tumors. However, serum alpha-lactalbumin was detected by radioimmunoassay in only 10% of women with infiltrating ductal carcinoma, indicating that this may not be a useful marker for the presence of breast cancer at least by the method employed. This discrepancy between serum and tissue alpha-lactalbumin may be due to the short half-life of serum alpha-lactalbumin or to the lack of secretion of the protein into the blood. There was no apparent relationship between the presence of estrogen receptors or serum prolactin and alpha-lactalbumin in the 27 mammary carcinomas investigated.

To investigate the cellular differentiation of adenoid cystic carcinomas (ACC), a comparative immunohistochemical study of 12 normal salivary glands and eight specimens of ACC was performed. Antibodies were used against S100 protein (S), keratins (K) of various molecular weights, vimentin (V), muscle-specific actin (A), epithelial-membrane antigen, human milk fat globules, and collagen type IV. A panel of four of these antibodies (SKVA) was identified as the most helpful in characterizing cells in normal salivary glands and ACC. The immunophenotypes depended on the histologic patterns of ACC. Cells in morphologically recognizable duct structures in the cribriform and trabecular areas expressed a phenotype similar to that of the intercalated duct. Cell layers around pseudocysts and occasional cellular islands had an immunophenotype suggesting myoepithelial-cell differentiation. The most clear cut epithelial/myoepithelial bilaminar differentiation was present in areas with a trabecular pattern, in which the layers facing the stroma and the central ductal elements had SKVA phenotypes of myoepithelial and ductal differentiation, respectively. In areas with a reticular pattern, most of the cells showed ductal differentiation. Many of the cells in the cribriform and basaloid regions were immunophenotypically undifferentiated. These results indicate that ACC consists of undifferentiated cells and of cells that are differentiating toward ducts, predominantly intercalated ducts, and toward myoepithelium. These findings support previous observations by electron microscope.

Six patients with malignant head and neck tumors are shown to have required electron microscopy for accurate diagnosis. In all of these tumors, there were ultrastructural features of cytodifferentiation that were not discernible by light microscopy, such as neurosecretory granules, desmosomes, cytoplasmic processes, tonofibrils, and myofilaments. Electron microscopy is helpful in the differential diagnosis of tumors in general, but its effectiveness is particularly apparent in small-cell "undifferentiated" tumors such as neuroblastoma, rhabdomyosarcoma, Ewing's sarcoma, undifferentiated squamous-cell carcinoma of the lymphoepithelioma type, and malignant lymphoma. It has also been helpful in the identification of amelanotic melanoma and spindle-cell carcinoma.

A 57-year-old man underwent a right pneumonectomy for a bronchogenic carcinoma following bronchoscopy and bronchial biopsy. The tumor was a polypoid mass arising from the lower lobe bronchus. Microscopically it was characterized by mononuclear cells mixed with randomly distributed multinucleated giant cells similar to those seen in giant-cell tumor of bone. Also found were portions showing typical squamous cell and spindle cell carcinoma. Based on the light and electron microscopic findings, we suggest that the current case represents a metaplastic squamous carcinoma showing mesenchymal cell differentiation. A hypothesis on the histogenesis of pleomorphic carcinomas was presented. The problems of histological diagnosis generated by such a tumor should be emphasized.

Fifteen malignant schwannomas were examined by light and electron microscopy. Five tumors arose in patients with neurofibromatosis and five were contiguous with a peripheral nerve (Group I). Five tumors met neither of these generally accepted diagnostic criteria but were light microscopically seen as compatible with malignant schwannoma when examined under light microscope (Group II). In the better differentiated areas of Group I lesions, long, overlapping, tightly packed cytoplasmic processes were parallel to homogeneous flocculent material, occasionally assuming a linear appearance suggesting basal lamina. In Group II, similar cytoplasmic processes were present but the extracellular material was less extensive and had a less obvious relationship to the plasma membrane. In neither group were fine intracytoplasmic filaments prominent. Malignant schwann cells are seldom as ultrastructurally differentiated as their benign counterparts. Nevertheless, within the context of well-studied light microscopy and the sampling error inherent in ultrastructural examination, electron microscopy can support the diagnosis of malignant schwannoma.

A broad ligament tumor with the histologic features of a female adnexal tumor of probable Wolffian origin, as described by Kariminejad and Scully, is reported. Although previous cases were thought to be benign, in the present case, hepatic metastasis occurred 6 years after resection of the primary. Cytologic features by light microscopy were more anaplastic than in the original cases. Ultrastructural findings were not specific for any normal adult or fetal mesonephric tissue previously studied by electron microscopy.

The histologic distinction between mesotheliomas and adenocarcinomas on mesothelial surfaces is sometimes difficult. High concentrations of hyaluronic acid in pleural and peritoneal fluids of mesothelioma patients have been reported. This paper describes results of glycosaminoglycan electrophoresis of papain digests of 7 mesotheliomas and 27 other tumors. Either all or almost all demonstrable glycosaminoglycans in mesotherliomas was hyaluronic acid. Control tissues had variable electrophoretic patterns showing other glycosaminoglycans in addition to hyaluronic acid. In the clinical context of differentiating mesotheliomas and adenocarcinomas in tissue samples, this technique seems simple, economical, and relatively specific.

A monoclonal antibody designated as T29/33, which recognizes a major cell surface glycoprotein of hematopoietic cells, was tested as a possible tool to distinguish lymphoma from nonhematopoietic tumors having a similar morphologic appearance by light microscopy. Each of the 40 lymphomas studied by immunofluorescence and/or immunoperoxidase on frozen sections revealed the presence of the corresponding antigen, whereas 110 control tumors of various nonhematopoietic origins were negative. It was concluded that the T29/33 antibody may be of value in resolving the diagnostic problems presented by tumors in which the differential diagnosis is between non-Hodgkin's lymphoma and nonlymphoid tumors.

The authors investigated the expression of keratin, carcinoembryonic antigen (CEA), and an epithelial marker derived from milk fat globule membranes in 12 mesotheliomas and 100 diverse adenocarcinomas with immunohistochemical methods. The authors employed a monoclonal antibody to keratin designated as AE1, as well as the following commercially available antisera: rabbit anti-whole human keratin, rabbit anti-CEA, and a monoclonal antibody to an epithelial factor designated as MFG-2. Expression of keratin was found in all the mesotheliomas and adenocarcinomas with antibody AE1 as well as with the rabbit antiserum; CEA was detectable in 65% of the adenocarcinomas but two mesotheliomas also reacted weakly. With antibody MFG-2, positive results were obtained in 85% of the adenocarcinomas and in none of the mesotheliomas. All of 64 (100%) breast-, lung- and ovary-derived adenocarcinomas immunostained positively with antibody MFG-2. This is of particular significance because pulmonary and ovarian adenocarcinoma frequently may be indistinguishable clinically and histologically from epithelial mesothelioma. The authors conclude that antikeratin antibodies are not useful in the distinction of adenocarcinoma from mesothelioma. Because of its greater sensitivity and specificity, MFG-2 is superior to CEA in this differential diagnosis.

Three cases of angiosarcoma in the stomach, small bowel, and colon, respectively, are reported. The gastric lesion occurred many years after historic evidence of cutaneous Kaposi's sarcoma. Two of the three lesions exhibited multifocality, and all demonstrated aggressive biologic courses. The varied histologic features of angiosarcoma make this diagnosis difficult to establish in an unusual location such as the gastrointestinal tract. The use of immunohistochemistry for a panel of antibodies, particularly the successful reaction with Factor VIII-related antigen, is important in evaluating light microscopically unusual tumors. A delicate framework of collagen type IV was elicited in each case, but ultrastructurally correlated with short segments of basal lamina in one case. This suggests that the absence of the traditional morphologic expression of basal lamina does not preclude its presence and emphasizes the sensitivity of immunohistochemistry. The lack of previously reported cases of gastrointestinal angiosarcoma may be because the tumor is underrecognized or because it is truly a rare occurrence.

Cholestasis is a common complication of total parenteral nutrition (TPN) in infants. A contributing factor to the hepatic dysfunction may be a contaminant of the TPN solution, such as aluminum, that accumulates in liver and may act as a hepatotoxin. To study the hepatic effects of aluminum, growing piglets were given daily intravenous injections of aluminum, 1.5 mg/kg, for 50 days; pair-fed controls were given heparinized saline. At sacrifice, liver and serum were obtained. Liver was analyzed for histopathology and for aluminum content and localization. The hepatocyte lysosomes of the experimental group showed aluminum peaks by x-ray microanalysis, whereas the control group did not. No differences in ultrastructure were noted between the two groups when examined by electron microscopy. Mean serum total bile acid levels (27.8 +/- 15.9 SD vs 6.3 +/- 1.5 mumol/liter, p less than 0.05), mean alkaline phosphatase (309 +/- 108 vs 180 +/- 27 IU/liter, p = NS), and mean hepatic copper content (24.8 +/- 4.5 vs 14.4 +/- micrograms/g dry weight, p less than 0.01), were elevated in the aluminum-loaded piglets, indicating that cholestasis may have been produced. Also, a small but significant reduction in serum levels of 25 hydroxy-vitamin D was found in the aluminum-loaded piglets, suggesting that vitamin D hydroxylation may be impaired. Inasmuch as lysosomal contents are excreted into the bile, aluminum accumulation in lysosomes may alter lysosomal function and possibly affect bile flow or content.

The authors investigated the presence and distribution of keratin in germ cell tumors using a rabbit-anti-keratin antiserum and a monoclonal antikeratin antibody--which is specific for keratin classes of 40, 50, and 56.5 kdaltons--by various immunohistochemical methods on frozen sections, alcohol-fixed, and formalin-fixed paraffin-embedded tissues. Thirty-four germ cell tumors were studied. These were the following: 18 seminomas, 10 embryonal carcinomas, 2 teratocarcinomas, 3 yolk sac tumors and 1 choriocarcinoma. All seminomas, including four poorly differentiated (so-called anaplastic seminomas), gave negative results, regardless of the method employed. Embryonal carcinoma, the epithelial component of the teratocarcinoma, the yolk sac tumors, and choriocarcinoma were at least focally positive for keratin. The monoclonal antibody provided a cleaner background and stronger staining than the rabbit-anti-total-human-epidermal-keratin antibody. Best results were obtained from fresh-frozen sections or alcohol-fixed, paraffin-embedded materials. Formalin-fixed, nonseminomatous tumors, when predigested with trypsin and incubated overnight with primary antibody, gave no false-negative results but staining was often focal. The authors' results agree with the reported absence of detectable keratin in primordial germ cells of the normal testis, and with prevailing concepts of the histogenesis of germ cell tumors. These results indicate that the presence or absence of keratin by immunocytochemical methods can be helpful in distinguishing seminoma from embryonal carcinoma.

Two cases of primary non-Hodgkin's lymphomas of the breast were studied with light and electron microscopy. By light microscopy, according to Rappaport's classification, one was a poorly differentiated lymphocytic lymphoma, the other a diffuse histiocytic lymphoma. However, the ultrastructural features of the latter were more consistent with transformed lymphocytes. The differential diagnosis with medullary and poorly differentiated carcinomas of the breast is discussed. The first case had rapid dissemination as did most of the cases reported in the literature.

Two cases of plexiform tumor of the uterus are presented. Both were incidental microscopic findings in leiomyomatous uteri and had the typical branching cords of small, polygonal cells with scanty cytoplasm and vesicular nuclei. Ultrastructurally the cells had features of smooth muscle differentiation such as actin-like filaments, dense bodies, peripheral dense plaques, pinocytotic vesicles, and incomplete basal lamina. Their close relationship to capillaries resembled the structure of glomus tumor and vascular leiomyoma and suggested histogenetic kinship among these neoplasms.

Two cases of spindle cell carcinoma (SCC) of the esophagus and skin, respectively, were studied by transmission electron microscopy. The tumor cells were closely associated with collagen fibrils and had abundant rough endoplasmic reticulum with dilated cisternae. Except for their irregular shape and atypical nuclei they resembled actively synthesizing fibroblasts. In addition to these features, some cells contained numerous tonofibrils and occasional well-developed desmosomes. A gradual transition to typical squamous cells was noted in the skin tumor. These findings suggest that the pseudosarcomatous component of SCC originates from mesenchymal metaplasia of squamous cells and that collagen is produced by these metaplastic cells.

A case of small cell carcinoma of salivary gland was studied by light and electron microscopy. Light microscopy showed sheets of small uniform cells with scanty cytoplasm and pyknotic nuclei. Electron microscopy showed two distinct cell types; an electron-lucent epithelial-appearing cell, and a denser cell with processes containing bundles of filaments and other features suggesting myoepithelial differentiation. Neurosecretory granules were absent. These findings support a salivary duct origin, and are evidence against a neurendocrine derivation for this tumor.

Sixteen cases of malignant fibrous histiocytoma are presented. Electron microscopy of 15 cases demonstrated fibroblast-like and mononuclear and multinucleated histiocyte-like cells. A small capillary was at the center of all storiform areas examined. Ultrastructural examination can be diagnostically useful within the context of a narrow differential diagnosis by conventional microscopy and the ability, by electron microscopy, to eliminate other mesenchymal cell types. In 13 cases, follow-up information was available from 18 months to 9 years following histological diagnosis. Five patients are alive and 8 patients have died, including two non-tumor related deaths. In 3 cases follow-up was less than 4 months. The biologic behavior of the tumor in this series was generally not related to histopathological parameters. The issue of histogenesis is largely unresolvable. Ultrastructural studies of various types of fibrous histiocytomas, suggesting cells of origin other than histiocytes, give credence to the concept that the histiocyte may represent a morphologic state of a given mesenchymal cell rather than a particular cell type.

Twenty-seven primary non-small cell (NSC) lung cancers were analyzed for alterations of protooncogenes by DNA hybridization techniques. Abnormalities were found in 56% of tumors including ten of 16 adenocarcinomas, three of nine squamous cell cancers and two of two larger cell cancers. Five protooncogenes were found to be commonly altered in tumors at frequencies between 12% and 60%. These were c-myc, c-myb, c-ras-Ha, c-erbB-1 and c-erb-B-2. Alterations in c-erbB-1 and c-erbB-2 correlated with histologic type of tumor and were more common in advanced cancers. Allelic deletions of c-ras-Ha or c-myb were frequently observed in primary tumors which recurred or progressed after surgery (five of six). Analysis of protooncogenes may provide insights into the pathogenesis of lung cancer and may aid in predicting clinical behavior.