Biomedical subjects

J W Said

Publications and source records attributed to J W Said.

At least 109 records · Page 6Linked to original sources

PubMed1987

Detection of estrogen receptors with monoclonal antibodies in routinely processed formalin-fixed paraffin sections of breast carcinoma. Use of DNase pretreatment to enhance sensitivity of the reaction.

This study describes an improved immunohistochemical method for the sensitive and specific identification of estrogen receptors (ERs) in paraffin sections from formalin-fixed and routinely processed breast carcinoma tissues, using DNase pretreatment to expose nuclear antigenic sites and commercially available immunoreagents (including monoclonal antibody) in kit form. Results were compared with dextran-coated charcoal cytosolic assay (DCC) and with conventional immunohistochemistry on frozen sections. Sensitivity and specificity for determinations on paraffin sections were 88% and 86%, respectively, and statistical analysis showed very good agreement between DCC and paraffin sections (kappa = 0.805). The DNase technic on paraffin sections allows excellent correlation between histologic characteristics and ER status and reduces DCC sampling error resulting from stromal dilution and tumor variability. This method offers a reliable and reproducible alternative when tissue is not suitable or unavailable for DCC or frozen tissue analysis and can be used for retrospective studies on stored tissue blocks.

Antibodies, Monoclonal↗

PubMed1986

Endothelial cell markers in vascular neoplasms: an immunohistochemical study comparing factor VIII-related antigen, blood group specific antigens, 6-keto-PGF1 alpha, and Ulex europaeus 1 lectin.

Markers for endothelial cells including Ulex europaeus 1 lectin, blood group A, B, and H, and the prostaglandin metabolite 6-keto-PGF1 alpha were evaluated in paraffin secretions from formalin-fixed benign and malignant vascular neoplasms using a variety of immunohistochemical techniques, and results compared with staining for factor VIII-related antigen. Staining for Ulex appeared more sensitive than factor VIII-related antigen in identifying poorly differentiated neoplasms including haemangiosarcomas and spindle cell proliferations in Kaposi's sarcoma. Staining for blood group related antigens correlated with blood group in all cases. Ulex europaeus 1 lectin was the only marker for endothelial cells in lymphangiomas.

6-Ketoprostaglandin F1 alpha↗

PubMed1986

Non-Hodgkin's lymphoma, multilobated B-cell type: report of nine cases with immunohistochemical and immunoultrastructural evidence for a follicular center cell derivation.

The clinical, pathologic, and immunologic features in nine cases of non-Hodgkin's lymphoma of the multilobated B-cell type are described. Clinical and phenotypic heterogeneity was observed in these B-cell neoplasms. A probable follicular center cell derivation for these cytologically unusual B-cell lymphomas is supported by antecedent histories of follicular center cell neoplasms in three cases; a focal nodular pattern in one case; the demonstration of peanut lectin (PNA) receptors, a marker for follicular center cells, on neoplastic multilobated B cells; and immunoultrastructural studies of nonneoplastic tonsillar cells that identified and characterized rare multilobated cells, immunoreactive for B1, B2, and Ia membrane antigens, a phenotype consistent with follicular center-type cells. Comparison of B- and T-cell multilobated lymphomas revealed that only immunologic studies accurately discriminated between these neoplasms.

Adolescent↗

PubMed1986

Keratoacanthoma and squamous cell carcinoma of the skin: immunohistochemical localization of involucrin and keratin proteins.

Fifteen keratoacanthomas and fifteen squamous cell carcinomas of the skin were examined by immunoperoxidase methods for involucrin and both 45- and 63-kilodalton keratins. Keratoacanthomas showed a relatively homogeneous staining pattern for involucrin; all cells except basal cells stained with mild to moderate intensity. Squamous cell carcinomas disclosed a highly irregular involucrin staining pattern with marked variation in staining intensity from cell to cell. Staining patterns for keratin proteins did not appear to distinguish between keratoacanthomas and squamous cell carcinomas. The 45-kilodalton keratin pattern showed diffuse staining within both keratoacanthomas and squamous cell carcinomas, and the 63-kilodalton keratin pattern consisted of focal staining, mostly of dyskeratotic cells. These results suggest that involucrin may serve as a diagnostic aid in differentiating between squamous cell carcinomas and keratoacanthomas. In addition, other lesions in the differential diagnosis of keratoacanthoma and squamous cell carcinoma were also examined for involucrin.

Adult↗

PubMed1986

Leu-M1 immunoreactivity in nonhematopoietic neoplasms and myeloproliferative disorders. An immunoperoxidase study of paraffin sections.

Leu-M1 is a differentiation antigen present on human myelomonocytic cells, which also has been identified in Reed-Sternberg cells and variants of Hodgkin's disease. This study further defines the tissue distribution of Leu-M1, with immunoreactivity observed for neoplastic cells in 14 of 28 formalin-fixed, paraffin-embedded nonhematopoietic neoplasms. With the use of monoclonal antibodies and an indirect immunoperoxidase technic, Leu-M1 was detected in adenocarcinomas of various sites (breast, lung, colon, thyroid, pancreas, and stomach), in squamous cell and transitional cell carcinomas, and in a small-cell anaplastic carcinoma. Evaluation of a wide variety of myeloproliferative disorders indicated that Leu-M1 effectively characterized mature and immature monocytic cells and myeloid cells at late stages of granulopoiesis, but it was not a reliable marker for early myeloid cells including blasts. Leu-M1 monoclonal antibodies are a useful diagnostic reagent, particularly in the assessment of lymphoproliferative disorders, but must be used with extreme caution and full awareness of its staining profile.

Antibodies, Monoclonal↗

PubMed1986

Gastroenteropancreatic neuroendocrine tumors. A histochemical and immunohistochemical study of epithelial (keratin proteins, carcinoembryonic antigen) and neuroendocrine (neuron-specific enolase, bombesin and chromogranin) markers in foregut, midgut, and hindgut tumors.

Thirty-four gastroenteropancreatic (GEP) neuroendocrine tumors were evaluated for expression of epithelial (keratin, carcinoembryonic antigen [CEA] and neuroendocrine (neuron-specific enolase, chromogranin, bombesin) markers, and results were correlated with histologic patterns and histochemical staining. Tumors of mixed pattern (insular or trabecular with glandular areas) predominated. CEA localization corresponded to staining for mucin, with polarized apical or lumenal staining in glandular areas. Four trabecular midgut carcinoids, however, revealed diffuse cytoplasmic staining for CEA. Staining for keratin proteins was present in 68% of tumors. Bombesin immunoreactivity was demonstrated in 60% of GEP neuroendocrine tumors, indicating that bombesin positive metastatic tumors may not be predominantly of pulmonary origin, as previously suggested. Chromogranin was a sensitive marker for identifying normal gastrointestinal neuroendocrine cells that were not demonstrated by staining for neuron-specific enolase. Chromogranin was present in most neuroendocrine tumors, but was absent from three of five rectal carcinoids in keeping with the distinctive profile of hormonal and silver staining in these tumors. All GEP neuroendocrine neoplasms expressed both neuroendocrine and epithelial markers, supporting their derivation from endodermal epithelium.

Bombesin↗

PubMed1986

Immuno-ultrastructural localization of B-cell-specific monoclonal antibodies B1 and B2.

Monoclonal antibodies B1 and B2 are thought to recognize B-lineage restricted antigens, and have been used to define stages of B-cell maturation and characterize B-cell lymphomas. Immunostaining on cryostat sections has revealed a puzzling dendritic or extracellular pattern of staining for B2 within germinal centers and neoplastic follicles. In this study B1 and B2 are localized precisely on hyperplastic and neoplastic lymphoid tissues using immuno-ultrastructural techniques on cryostat sections, cell suspensions, and cell monolayers. B1 and B2 were localized to cell surfaces, including microvillous surface projections, on small and large transformed normal and neoplastic B lymphocytes. B2, in addition to staining in lymphoid cells, was localized to anastomosing cytoplasmic processes of dendritic histiocytes. These findings explain the apparently extracellular localization of B2 in cryostat sections and indicate that patterns of staining for B2 may represent a combination of staining on lymphoid cells and dendritic histiocytes.

Antibodies, Monoclonal↗

PubMed1986

Immunoultrastructural and morphometric analysis of B lymphocytes in human germinal centers. Evidence for alternate pathways of follicular transformation.

Monoclonal antibodies to B-cell differentiation antigens B1, B2, C3b, and Ia were used for ultrastructural characterization of B lymphocytes undergoing follicular transformation in human germinal centers. Morphologic alterations and morphometric parameters including form factor (FF) and nuclear contour index (NCI) were evaluated. Antibodies to B1, Ia, and C3b revealed uninterrupted linear surface membrane staining in B cells at various stages of transformation, while staining for B2 appeared as aggregates of gold particles localized to sites of antigen expression along the cell membrane. B cells with highly irregular or convoluted nuclei (NCI greater than 6.5) formed 3% of follicular lymphocytes and may explain the derivation of rare follicular center cell lymphomas with marked nuclear irregularity which mimic T-cell lymphomas histologically. Cleaved cells (NCI greater than or equal to 4.5) comprised 48% of the cellular population and were present at all stages of transformation. Results of morphometric studies suggest that small cleaved cells (centrocytes) and noncleaved cells transform to large lymphoid cells (centroblasts) along parallel lines, and without following the sequential differentiation pathway suggested by Lukes and Collins.

Antibodies, Monoclonal↗

PubMed1986

Diagnostic immunopathology.

The application of immunologic techniques to tissue sections has added a new dimension to the investigation and classification of various processes. Virtually every section of diagnostic pathology has been enhanced by using specific monoclonal antibodies or polyclonal antiserum. Neoplasms formerly diagnosed as poorly differentiated or anaplastic may be precisely identified as to their origin through the use of specific membrane or cytoplasmic markers. Other cellular products, including viruses, hormones, enzymes or highly specific proteins, are also available to study neoplastic and nonneoplastic processes. New and more specific reagents are regularly becoming available for the diagnostic repertoire of pathologists. We present some of the principles of diagnostic immunopathology to show the scope and importance of the techniques.

Adult↗

PubMed1985

Involucrin, a marker of squamous and urothelial differentiation. An immunohistochemical study on its distribution in normal and neoplastic tissues.

Involucrin is a soluble protein precursor of the cross-linked envelope present in the submembranous zone of human stratum corneum, and subsequently demonstrated in stratified squamous epithelia. The immunoperoxidase technique was used to assess the distribution of involucrin in 107 normal and 318 abnormal tissues. With few exceptions, involucrin was restricted to squamous epithelia, urothelium, some skin appendages and thymic Hassall's corpuscles. In normal squamous epithelium and normal urothelium, staining was most intense in the superficial layers where it was concentrated at the cell periphery and gradually decreased toward the basal layer. This orderly staining pattern was maintained in benign squamous and urothelial lesions and in grade I papillary urothelial carcinomas. Higher grade papillary urothelial carcinomas, infiltrating urothelial and squamous carcinomas, and in situ urothelial and squamous carcinomas demonstrated abnormal staining patterns for involucrin that are described. Foci of squamous differentiation in adenocarcinomas and other epithelial malignancies stained intensely for involucrin. Brenner tumours of the ovary and Walthard rests of the fallopian tube, lesions of uncertain histogenesis but possibly urothelial-related, also stained for involucrin. Results of this study suggest that involucrin is a sensitive and specific marker for squamous and urothelial differentiation, staining patterns for involucrin may be helpful in distinguishing benign from malignant urothelial and squamous lesions, and presence of involucrin may be helpful in determining the histogenesis of selected lesions.

Carcinoma in Situ↗

PubMed1985

Immunoreactive neuron-specific enolase, bombesin, and chromogranin as markers for neuroendocrine lung tumors.

Sixty-four lung tumors were evaluated for the presence of immunoreactive neuron-specific enolase (NSE), bombesin (Bn), and chromogranin (Cg) to assess their value as markers for neuroendocrine cells in the histologic diagnosis of pulmonary neoplasms. Staining was correlated with the presence and density of neurosecretory granules (number of neurosecretory granules per unit cytoplasmic cross-sectional area) as determined by planimetry on electron micrographs. The cytoplasmic density of neurosecretory granules was significantly greater in the carcinoid tumors than in the small cell carcinomas (P less than 0.001). Neuron-specific enolase was localized in all of the neuroendocrine granule-bearing tumors but was also present in 57 per cent of the nonneuroendocrine carcinomas. Bombesin was present in 68 per cent of the neuroendocrine tumors and in less than 1 per cent of the nonneuroendocrine tumors. Staining for Cg appeared to correlate with the density of neuroendocrine granules, with staining in carcinoid tumors but no staining in small cell anaplastic carcinomas. A panel of antibodies may be required for the reliable identification of neuroendocrine lung tumors by immunohistochemical techniques.

Bombesin↗

PubMed1985

Involucrin in intraepithelial and invasive squamous cell carcinomas of the cervix: an immunohistochemical study.

Immunohistochemical staining for involucrin, a cytoplasmic protein synthesized during squamous maturation, was assessed in histologic sections from hysterectomy and cone biopsy specimens from patients with cervical neoplasia. In normal and condylomatous squamous epithelium, diffuse cytoplasmic staining was seen in the suprabasal layers, with no staining of the basal cells. Staining was absent in two cases of cervical intraepithelial neoplasia (CIN), grade III, in which the lesions were composed entirely of undifferentiated cells and markedly decreased in cases involving large numbers of basal cells. In 19 of 23 cases (83 per cent) of CIN, however, focal staining for involucrin was seen in large differentiated cells in the more superficial layers, and in two cases of keratinized CIN diffuse suprabasal staining was observed. Similarly, strong staining for involucrin was present in differentiated areas in one case of microinvasive squamous cell carcinoma and in 93 per cent of cases of infiltrating squamous cell carcinoma. These findings suggest that involucrin is a marker for maturation in cervical squamous epithelial neoplasms. Patterns of immunohistochemical staining for involucrin in keratinized dysplasia and differentiated squamous carcinomas should be taken into consideration if loss of involucrin staining is used as a criterion for neoplastic transformation of cervical epithelium, as has been proposed.

Carcinoma, Squamous Cell↗

PubMed1985

Chromogranin as a marker of neuroendocrine cells in cytologic material--an immunocytochemical study.

Cytologic material, including fine-needle aspirations, bronchial brushings, body cavity fluids, and tissue imprints from 39 neuroendocrine and 26 nonneuroendocrine tumors, was stained for chromogranin by the immunoperoxidase technic. Our results suggest that chromogranin is useful in identifying primary as well as metastatic neuroendocrine lesions from a variety of body sites. All lymphoid proliferations and carcinomas, including small-cell anaplastic carcinomas of the lung (oat-cell carcinomas), were negative. Chromogranin appears to be a useful marker in diagnostic cytology. This technic can be applied to routinely prepared cytologic material.

Bronchial Neoplasms↗

PubMed1985

Demonstration of myoglobin and CK-M in myocardium. Comparison of five fixation methods and three immunohistochemical techniques.

The results of immunohistochemical staining vary depending on the tissue, fixative, antigen-antibody system, and immunohistochemical staining methods used. The purpose of this study was to evaluate the effect of different methods of fixation, different antigen-antibody systems, and different immunohistochemical methods on immunohistochemical staining of myocardium. Samples of normal fresh canine myocardium from six dogs were fresh frozen and fixed in 10% neutral buffered formalin, Bouin's, Bayley's and Carnoy's fixatives. Immunohistochemical staining for myoglobin and creatine kinase M was performed using the ABC (avidin-biotin complex) and indirect peroxidase-antiperoxidase (PAP) techniques. Tissues fixed in formalin showed the most intense specific staining for both antigens with the least background and nonspecific staining. All other fixation methods and frozen section techniques gave a more variable degree of specific positive staining and substantial background staining and/or nonspecific staining. ABC and PAP techniques gave similar results with both antigen-antibody systems and with each fixation method. Thus, no differences in specificity or sensitivity were observed between ABC and PAP techniques. Differences in staining intensity and pattern were related primarily to differences in fixation methods.

Animals↗

PubMed1985

Hodgkin's disease, lymphocyte predominance type, nodular--a distinct entity? Unique staining profile for L&H variants of Reed-Sternberg cells defined by monoclonal antibodies to leukocyte common antigen, granulocyte-specific antigen, and B-cell-specific antigen.

Using monoclonal antibodies to leukocyte common antigen, granulocyte-related antigen, and B-cell specific antigens, L&H variants of Reed-Sternberg (R-S) cells in Hodgkin's disease, lymphocyte predominance type (nodular), exhibited a unique staining profile as compared with R-S cells of other histologic types. L&H variants were strongly immunoreactive for leukocyte common antigen, as defined by monoclonal antibodies PD6/27 and 2B11; whereas other types of R-S cells were negative or rarely positive. R-S cells and variants in 69 cases of Hodgkin's disease of nodular sclerosis (41), mixed cellularity (25) or lymphocyte depletion (3) types, were consistently strongly immunoreactive for Leu-M1, a granulocyte-related antigen, while L&H variants were uniformly nonreactive (4 cases). B-cell specific antigens, detected by three pan-B-cell monoclonal antibodies, were observed only for L&H variants. These observations suggest that L&H variants of R-S cells represent a distinct type of transformed cell, possibly of B-cell origin, and do not share a common lineage with other types of R-S cells. These studies provide further evidence that Hodgkin's disease, lymphocyte predominance type, nodular, may represent a distinct entity.

Antibodies, Monoclonal↗

PubMed1985

Localization of alpha-1 acid glycoproteins in human myocardium.

alpha-1 acid glycoproteins become elevated in the patient's serum within a few hours after an acute myocardial infarction. Previous reports have suggested a correlation between the magnitude of this elevation and infarct size as estimated by enzyme markers. Correlation has also been observed between the mortality following infarction and appearance of elevated alpha-1 acid glycoproteins. We now report that these glycoproteins are detectable in normal myocardium using immunohistochemical techniques. Diminished amounts are observed in necrotic tissue in acute myocardial infarcts. Ultrastructural localization by immunoelectron microscopy using sections of normal myocardium established the presence of alpha-1 acid glycoproteins primarily at the cell surface in the region of the sarcolemma. The observations suggest that myocardium may directly contribute to the elevation of serum alpha-1 acid glycoproteins after an infarct, and the assessment of these components may serve as an additional serum marker.

Animals↗

PubMed1985

Leu-M1--a marker for Reed-Sternberg cells in Hodgkin's disease. An immunoperoxidase study of paraffin-embedded tissues.

Monoclonal antibody to Leu-M1, a granulocyte-related differentiation antigen, represents a highly effective reagent for detection of diagnostic Reed-Sternberg (R-S) cells and variants in paraffin-embedded tissues of Hodgkin's disease. In 69 of 73 cases of Hodgkin's disease (41 nodular sclerosis, 25 mixed cellularity, 4 lymphocyte predominance, and 3 lymphocyte depletion types), R-S cells were strongly immunoreactive for Leu-M1. Four cases of lymphocyte predominance Hodgkin's disease (nodular) were uniformly nonreactive for Leu-M1. In most of the positive cases (57/69, 83%), the majority (60-90%) of R-S cells and variants exhibited immunoreactivity for Leu-M1. A characteristic staining pattern included granular and/or vesicular cytoplasmic immunoreactivity, often with a prominent globular paranuclear reaction product, and membrane staining with highly irregular cytoplasmic borders. Evaluation of B-cell (37 specimens), T-cell (20 specimens), and true histiocytic (3 specimens) neoplasms and a case of mastocytosis revealed immunoreactivity for Leu-M1 only in 1 B-cell and 4 T-cell malignancies. The staining patterns in these cases, however, clearly differed from that observed for R-S cells. Studies of nonneoplastic lymphoid tissues (38 total) demonstrated that lymphoid cells were typically nonreactive; histiocytes revealed variable reactivity for Leu-M1. Occasional histiocytes of the sinusoidal network of lymph nodes, particularly in toxoplasmic lymphadenitis, exhibited a staining pattern (membranous/cytoplasmic/paranuclear) similar to that observed for R-S cells. Leu-M1 represents a potentially helpful diagnostic discriminant in the assessment of Hodgkin's disease and its distinction from non-Hodgkin's lymphomas and other lymphoid proliferations.

Antibodies, Monoclonal↗

PubMed1984

Identification and localization of creatine kinase B and M in normal, ischemic and necrotic myocardium. An immunohistochemical study.

We utilized immunoperoxidase methods to study the distribution of CK-B and CK-M in normal, ischemic and necrotic myocardium. Human myocardium was obtained from autopsy (n = 10) and surgery (n = 16). Cardiac tissue from 22 dogs with experimental myocardial infarction induced by closed-chest coronary balloon occlusion and four dogs with myocardial ischemia without necrosis induced by a 50% reduction in left main coronary artery blood flow for 3 h were studied. Duration of occlusion was 45 min (n = 2), 3 h (n = 8), 5 to 6 h (n = 7), 15 to 24 h (n = 5). Highly purified anti-CK-B and M were prepared in our laboratory and obtained commercially. In all cases, control experiments were performed. Microscopically normal human and dog myocardium uniformly stained for CK-B and CK-M. Necrotic myocardium from patients with acute infarcts (10 to 24 h old) showed markedly reduced immunostaining. In dogs with 3 to 24 h occlusion immunostaining was significantly reduced for both CK-B and CK-M in regions confirmed to be necrotic by triphenyl tetrazolium chloride (TTC) and H & E staining. Myocardial necrosis was confirmed in the 3-h infarcts by electron microscopy (EM). In the four dogs with a 50% reduction in left main flow for 3 h, ischemia was demonstrated by glycogen loss in periodic acid-Schiff stained-sections; but there was no evidence of necrosis by EM or TTC, and there was no loss of immunostaining evident for CK-B and CK-M. Thus, using immunoperoxidase techniques, CK-B and CK-M were visualized in normal and ischemic myocardium, with decreased staining in necrotic tissue. These findings indicate that cell death is necessary for the demonstration of CK-M and CK-B loss from the myocardium by this technique.

Animals↗