Multivariate analysis of prognostic features in malignant pleural effusions from breast cancer patients.

To identify prognostic indicators in breast cancer patients with malignant pleural effusions, we analyzed the cytopathologic features of 57 fluids representing the first pathologic diagnosis of a distant metastasis in these women. The specimens were analyzed prior to reviewing the clinical records. The median survival of 55 patients who died of the disease was five months following the effusion diagnosis (range, 1-114). Univariate analysis identified three cytopathologic features that correlated with relatively prolonged survival: arrangement of tumor cells in spheroids, slight nuclear atypia and low mitotic rate. Women whose tumor cells formed spheroids survived a median of 24.5 months as compared to 4 months for women with all other architectural patterns (P = .004). Multivariate analysis revealed that slight nuclear atypia and low mitotic rate strongly correlated with spheroid formation. Since breast cancers that form spheroids in effusions portend a relatively favorable prognosis, we recommend that cytopathologists comment on this pattern when reporting on metastatic breast carcinoma in pleural fluids.

Immunostaining of small cytologic specimens. Facilitation with cell transfer.

Immunocytochemical study of cytologic specimens is often limited by the number of slides containing diagnostic cells. This study examined the effectiveness of transferring cells from a single smear to multiple slides in order to perform a battery of immunocytochemical stains on limited material. Immunostaining performed on four effusions and five fine needle aspirates yielded the expected results for most of the antibodies commonly employed in diagnostic pathology. Background staining was generally low following cell transfer, and the morphology of the cells was preserved. These results suggest that cell transfer in combination with immunocytochemistry may prove useful in the cytologic diagnosis of malignant lymphoma, neuroendocrine neoplasms, prostatic and mammary adenocarcinoma, and other malignant tumors.

p53 expression in precancerous gastric lesions: an immunohistochemical study of PAb 1801 monoclonal antibody on adenomatous and hyperplastic gastric polyps.

To determine the chronology of p53 mutation in the gastric carcinogenic sequence, we studied p53 overexpression in premalignant lesions, including 17 adenomatous polyps (10/17 surrounded by intestinal metaplasia and 11/17 harboring foci of adenocarcinoma or severe dysplasia), and 18 hyperplastic polyps (4/18 with focal adenomatous changes). Immunohistochemistry with PAb 1801 monoclonal antibody was performed on archival material; p53 nuclear staining was seen in 10/17 adenomas, but was limited to the foci of adenocarcinoma in three cases. Five adenomas with foci of severe dysplasia or carcinoma were nonreactive. Intestinal metaplasia, normal gastric mucosa, and 14/18 hyperplastic polyps were nonreactive. p53 Reactivity observed in four hyperplastic polyps was limited to adenomatous foci. These results suggest that p53 overexpression occurs in dysplastic epithelium of precancerous gastric lesions. Its absence in chronic atrophic gastritis with intestinal metaplasia suggests it is a relatively late event in the gastric carcinogenic sequence.

Blood group antigens in Barrett's esophagus and associated adenocarcinomas.

A panel of monoclonal antibodies (MoAbs) against BGAg A, B, and H and Lewis a, b, X, and Y (Le a, Le b, X, Y) was used to characterize and evaluate the blood group antigens (BGAg) immunohistochemistry in Barrett's esophagus (BE) and associated adenocarcinomas (ACA). Thirteen cases of ACA arising in metaplastic columnar epithelium of the distal esophagus were reviewed (12 men; 1 woman; mean age: 59). Twelve cases had juxtaposed BE of different types with low-grade dysplasia in five and high-grade dysplasia in three. ACAs were classified as well differentiated (one case), moderately differentiated (three cases), and poorly differentiated (four cases). Five cases had mixed features. Loss of BGAg H was recorded in four ACA and in one adjacent BE. Loss of BGAg B was noted in three ACAs, but in only one case in the adjacent BE. Incompatible expression of BGAg A and B was present in one case each. Anomalous expression of Le a was seen in seven ACAs and adjacent BE. Loss of Le b was noted in six ACAs. Deletion of X was seen in four ACAs and in BE. Loss of Y was present in the only nonsecretor patient. We conclude that significant alterations of BGAg occur in BE and in associated ACAs. Recognition of these antigenic alterations warrants further investigation to define a role for BGAg immunohistochemistry in the surveillance of BE patients at risk of developing ACA.

Epithelial cysts in the central nervous system, characteristic expression of cytokeratins in an immunohistochemical study.

Nineteen epithelial cysts in the central nervous system including six colloid cysts of the third ventricle, seven Rathke's cleft cysts in the sella, two enterogenous cysts in the posterior fossa, two epithelial cysts in the spinal canal and two neuroectodermal cysts in the cerebrum were examined immunohistochemically for expression of intermediate filament proteins-simple type, stratified type and skin type cytokeratins and GFAP. Colloid cysts of the third ventricle, Rathke's cleft cysts in the sella and epithelial cysts in the spinal canal expressed complex type cytokeratins while enterogenous cysts and neuro-ectodermal cysts showed only simple type cytokeratins. In addition, Rathke's cleft cysts expressed GFAP in occasional lining cells. The characteristic composition and distribution of cytokeratins in various kinds of epithelial cysts in the central nervous system are demonstrated and discussed with regard to their origins.

Morphologic diversity of long bone adamantinoma. The concept of differentiated (regressing) adamantinoma and its relationship to osteofibrous dysplasia.

A review of the clinical, radiologic, and histologic features of 25 cases of long bone adamantinoma is presented. To answer some questions concerning the nature of these tumors, relevant tissue markers were analyzed in seven cases using immunohistochemical assays. This study confirmed the epithelial nature of long bone adamantinomas irrespective of their wide-ranging morphologic patterns that can mimic tumors of various origin. On the basis of distinct radiologic, histologic, and immunohistochemical patterns, a new type of adamantinoma termed "differentiated adamantinoma" could be distinguished from the classic long bone adamantinomas. The diagnostic features characteristic of the differentiated adamantinoma include: patient age (first two decades), intracortical location of the entire lesion, uniform predominance of an osteofibrous dysplasia-like pattern, and scattered positivity of epithelial elements for cytokeratin. We postulate that the predominance of an osteofibrous dysplasia-like pattern in differentiated adamantinoma is the result of a secondary reparative process overgrowing matured and regressing tumor tissue. It is possible that this process may lead to the total elimination of recognizable tumor cells from the lesion. Therefore, osteofibrous dysplasia (ossifying fibroma) of long bones, which has a similar anatomic location, age distribution, and radiologic appearance as differentiated adamantinoma, may, in some cases, represent the evolution of an underlying adamantinoma. Our analysis suggests that long bone adamantinoma could be another member of the unique family of tumors that may regress spontaneously.

A comparative immunocytochemical study of human cerebellar cortex in X-chromosome-linked copper malabsorption (Menkes' kinky hair disease) and granule cell type cerebellar degeneration.

A comparative immunocytochemical study on the cerebellar cortex with X-chromosome-linked copper malabsorption (X-cLCM) and granule cell type cerebellar degeneration (gc-CD) was carried out by using specific monoclonal antibodies to synaptophysin (SY) and glial fibrillary acidic protein (GFAP). In X-cLCM cases, marked depletion of SY-immunoreactivity (IR) and reduction in number of SY-positive glomeruli were seen in the molecular and granular layers, respectively. Abnormal Purkinje cells occasionally showed moderately strong SY-IR having a fine granular pattern. Proliferation of GFAP-positive cells was observed in the granular and Purkinje cell layers. In the gc-CD case, SY-positive materials were coarsely distributed in a less dense fashion in the molecular layer as compared to a normal control. Purkinje cell perikarya did not show SY-IR. In the gc-CD granular layer, SY-IR appeared to have a coarsely punctate pattern, whereas immunoreactive glomeruli were almost completely absent. A number of GFAP-positive Bergmann cells was observed in the Purkinje cell layer and their fibres were densely and irregularly distributed in the molecular layer, whereas the granular layer was devoid of GFAP-positive cells. We present an immunocytochemical study of the X-cLCM and gc-CD cerebellar cortices, discuss the possible pathogenic mechanisms occurring in these diseases and discuss the usefulness of the SY-immunostaining technique for visualization of axon terminal involvement in these pathological conditions.

Testicular atrophy in homosexual AIDS patients: an immune-mediated phenomenon?

An immunopathologic analysis of the testes of 20 homosexual acquired immune deficiency syndrome (AIDS) patients was performed to investigate the hypothesis that the testicular atrophy of these patients represents an immune-mediated process. The findings were compared to those in a control group of heterosexual men without AIDS. The testes of the homosexual AIDS patients showed an overall lower degree of spermatogenesis with more prominent interstitial inflammation and thickening of the tubular basement membrane. However, direct immunofluorescence and immunoperoxidase studies failed to demonstrate significant differences in the prevalence of immune complex deposits along the tubular basement membrane in the two study groups. No electron-dense deposits were demonstrated in cases examined ultrastructurally. Therefore, this study does not support the hypothesis that the testicular atrophy of homosexual AIDS patients is a manifestation of autoimmune orchitis. Further studies are needed to elucidate the pathogenesis of the testicular atrophy in AIDS, as this appears to be a specific manifestation of the disease.

An immunohistochemical investigation of the human neostriatum in Huntington's disease.

The neostriatum of 7 autopsied patients with Huntington's disease (HD) was examined immunohistochemically using purified antibody against calcineurin, which may be present only in the medium-size spinous neurons of the mammalian striatum. This study revealed a marked loss of calcineurin-positive cells in the caudate nucleus and the putamen in all HD patients, compared with control subjects, and there was some variation among the HD patients. Four HD patients showed significantly lower density of calcineurin-positive cells in the caudate nucleus than in the putamen. The remaining calcineurin-positive cells in the caudate nucleus and the putamen had a mosaic-like pattern, demonstrating a subregional difference in distribution. This finding suggests that there are subregional as well as compartmental differences in the vulnerability of the calcineurin-positive cells in the striatum of patients with HD.

Focal accumulation of phosphorylated neurofilaments within anterior horn cell in familial amyotrophic lateral sclerosis.

Distribution of phosphorylated neurofilament proteins within anterior horn cells in three cases of familial and six cases of sporadic amyotrophic lateral sclerosis (ALS) and ten control cases were investigated by using a monoclonal antibody. Two distinct staining patterns of perikarya were observed; (1) homogeneous pattern; either the entire or a part of the perikaryon was immunostained homogeneously (homogeneously diffuse or partial pattern); (2) focal pattern: perikarya contained very distinct, inclusion-like focal accumulation of immunoreactive products of various morphologies such as round, ring-shaped, cord-like, tube-like and more irregular shapes. The homogeneous pattern was found in all three groups but was most common in sporadic ALS. On the other hand, the focal pattern was seen almost exclusively in familial ALS. The focal accumulation of neurofilaments appears at least in part to be related to the Lewy body-like hyaline inclusion which is known to contain neurofilaments. In addition, cord-like swellings of neurites in familial ALS also showed focal neurofilament accumulation. These observations suggest that the focal accumulation of phosphorylated neurofilaments is characteristic of familial ALS, although it may not be specific to the entity. The pathological process(es) producing the neurofilamentous abnormality may play an important role in anterior horn cell degeneration in familial ALS.

Calcineurin immunoreactivity in striatonigral degeneration.

The basal ganglia (including substantia nigra) of two patients with striatonigral degeneration, who had clinical histories of Parkinson's disease, were studied immunohistochemically using a purified antibody to calcineurin (CaN). Marked loss of CaN-immunoreactive neurons in the putamen and neuromelanin-pigmented neurons in the zona compacta of the substantia nigra was seen in both cases. A small number of CaN-immunoreactive neurons remained dispersed in "clusters" or "islands" in the medial portion of the putamen. In one case there was loss of CaN-immunoreactive neurons in the caudate nucleus to a lesser degree than that in the putamen. In addition, both cases showed marked depletion of CaN-immunoreactive afferent nerve fibers in the external and internal segments of the globus pallidus and the zona reticulata of the substantia nigra. This report emphasizes the usefulness of the CaN-immunostaining technique for assessing the striatal efferents in human basal ganglia, and shows the neuropathological changes in the basal ganglia with striatonigral degeneration which were not possible to ascertain with previous techniques.

Immunohistochemical visualization of afferent nerve terminals in human globus pallidus and its alteration in neostriatal neurodegenerative disorders.

The afferent nerve terminal in the human globus pallidus, which receives the projection nerve fibers from both the striatum and the subthalamic nucleus, were clearly visualized immunohistochemically using antibodies to calcineurin, synaptophysin, Met-enkephalin (MEnk) and substance P (SP). In normal control case, MEnk and SP-like immunoreactivities were densely localized in the external and internal pallidal segments, respectively, whereas calcineurin and synaptophysin were distributed throughout the globus pallidus. Calcineurin, synaptophysin, MEnk and SP-like immunoreactive peroxidase products decorated most of the long radiating dendrites and the cell bodies of the pallidal neurons. In the cases with Huntington's disease (HD) and striatonigral degeneration (SND), marked loss of calcineurin, MEnk and SP-like immunoreactivities was seen in the globus pallidus corresponding to areas of striatal neurodegeneration, whereas synaptophysin immunoreactivity remained in areas which revealed almost complete loss of calcineurin, MEnk and SP-like immunoreactivities. Calcineurin, MEnk and SP, which show difference in their localization patterns, may provide reliable markers for the striatal efferent nerve terminals, and synaptophysin for the entire pallidal afferent nerve terminals. This report demonstrates the distribution patterns of these neurochemical molecules in the globus pallidus with HD and SND.

Distribution of epithelial membrane antigen in normal and neoplastic human ependyma.

The ependyma and choroid plexus of 23 normal brains and 20 ependymal tumors were examined immunohistochemically for expression of epithelial membrane antigen (EMA) using a specific monoclonal antibody. The ependyma of normal brains showed three patterns of immunoreactivity: membrane immunoreactivity confined to the luminal surface; irregular punctate intracytoplasmic immunoreactivity in the subependymal layer; and spherical and ring-like intracytoplasmic immunoreactivity in the subependymal layer. Of 13 differentiated ependymomas 11 reflected the immunoreactive patterns of normal ependyma. The anaplastic ependymomas and ependymoblastomas had no immunoreactivity. Our results indicate that EMA has a highly selective distribution in the ependyma, and is a marker for differentiated ependymoma.

Thrombospondin mediates the cytoadherence of Plasmodium falciparum-infected red cells to vascular endothelium in shear flow conditions.

Cerebral malaria is thought to involve specific attachment of Plasmodium falciparum-infected knobby red cells to venular endothelium. The nature of surface ligands on host endothelial cells that may mediate cytoadherence is poorly understood. We have investigated the effects of soluble thrombospondin, rabbit antiserum raised against thrombospondin, and human immune serum on cytoadherence of parasitized erythrocytes in ex vivo mesocecum vasculature. Preincubation of infected red cells with soluble thrombospondin or human immune serum inhibits binding of infected red cells to rat venular endothelium. Infusion of the microcirculatory preparation with rabbit antithrombospondin antibodies before perfusion of parasitized erythrocytes also resulted in decreased cytoadherence. In addition, incubation of infected cells with human immune sera obtained from malaria patients significantly inhibited the observed cytoadherence. Our results indicate that thrombospondin mediates binding of infected red cells to venular endothelium and may thus be involved in the pathogenesis of cerebral malaria.

Prostatic carcinoma with endocrine features. A report of a neoplasm containing multiple immunoreactive hormonal substances.

A case of prostatic carcinoma with the cellular patterns of an adenocarcinoma and carcinoid tumor is reported. The tumor contained ultrastructural dense core neuroendocrine granules, and immunoperoxidase staining revealed prostatic acid phosphatase, prostatic-specific antigen, chromogranin, neuron-specific enolase, serotonin, adrenocorticotrophic hormone (ACTH), somatostatin, parathormone, calcitonin, bombesin, and glucagon but no insulin. The patient had exhibited hypercalcemia that may have been related to hormone production by the tumor. The literature on the endocrine aspect of the prostate and its tumor is reviewed.

Disseminated aspergillosis and pacemaker endocarditis.

Clinical and pathologic findings in a 65-year old woman with fever of unknown origin are described in this report. Generalized aspergillosis with endocarditis was demonstrated at autopsy. The patient had no recognized risk factors for the development of fungal infection. A functional transvenous pacemaker lead, inserted 2 years previously, was completely encased in a large infected thrombus and may have been the initial site of infection. Septicemia and endocarditis are rare but well-described complications of cardiac pacing, and should be considered in the differential diagnosis of fever of unknown origin in patients with pacemakers.

Keratosis punctata of the palmar creases.

We report on a 31-year-old man with keratosis punctata (punctate hyperkeratosis) of the palmar creases. We examined a lesion ultrastructurally and were unable with routine electron microscopic techniques to note any significant abnormalities.