Immunohistologic demonstration of abnormal colonic crypt cell kinetics in ulcerative colitis

A monoclonal antibody, Ki-67, that reacts with cells in the proliferative phases (G1, G2, S, and M) of the cell cycle was used in an immunohistochemical labeling reaction to examine the colonic crypt epithelium in active ulcerative colitis, inactive ulcerative colitis, and normal mucosa. The proportions of labeled cells in the lower two thirds (proliferative zone) and in the upper quarter of the crypt were determined. The proportions of Ki-67-positive crypt epithelial cells in both the proliferative zone and the upper crypt were higher in biopsy specimens from patients with active ulcerative colitis than from patients with normal mucosa or with inactive ulcerative colitis. In inactive ulcerative colitis the proportion of Ki-67-positive epithelial cells in the proliferative zone of the crypt was higher than in normal mucosa. These results are similar to those obtained in studies using tritiated thymidine to determine the proportion of cells in the DNA-synthesizing thymidine to determine the proportion of cells in the DNA-synthesizing phase of the cell cycle and suggest that immuno-histochemical staining with Ki-67 may be a practical method for measuring the proliferative activity of epithelial cells in patients with ulcerative colitis and other disorders of the gastrointestinal tract.     

Angioimmunoblastic lymphadenopathy with dysproteinemia. Immunohistologic and ultrastructural studies.

Immunoblasts of the B-cell line and immunoblasts of the T-cell line have very similar light-microscopic appearances but different immunologic compositions. The immunoblasts of a patient with angioimmunoblastic lymphadenopathy with dysproteinemia were found to bear surface immunoglobulin and complement receptor. They lacked receptor for cytophilic antibody. Surface villous projections were demonstrated by scanning electron microscopy. These features strongly indicate their B-cell origin.     

Immunohistologic labeling of murine endothelium.

BACKGROUND: Reliable identification of endothelial cells is a prerequisite for understanding vascularity changes in many cardiovascular diseases and therapeutic interventions. With the rising use of mouse models of disease and genetic manipulation, a consistent system to label murine endothelial cells in normal and diseased tissues would be an invaluable tool. METHODS: Immunohistologic and histochemical methods were used to label endothelial cells in normal and infarcted heart as well as small intestine. Isolectin B(4) or antibodies to thrombomodulin, vWF, Tie-2, VE-cadherin, CD34, and CD31 were used to immunostain tissues fixed in either 4% formaldehyde (prepared fresh from powdered paraformaldehyde, PF), methyl Carnoy's (MC), zinc (Zn) (all paraffin-embedded), or frozen sections. In addition, we used beta-galactosidase histochemistry in frozen sections from the Tie-2/beta-galactosidase transgenic mouse, in which the lacZ reporter gene is driven by the endothelial-specific Tie-2 promoter. RESULTS AND CONCLUSIONS: Immunohistologic localization of endothelial cells was best accomplished using anti-CD31 in Zn-fixed, paraffin-embedded tissues. Antithrombomodulin staining also worked in Zn-fixed tissues as well as frozen tissues, but additional steps were required to reduce background. The beta-galactosidase reporter system also marked endothelial cells in frozen Tie-2 transgenic mouse tissues; however, this system required careful control of fixation and optimization of histochemistry and was not amenable to double labeling. Lectin staining gave complete labeling of endothelial cells but cross-reacted intensely with macrophages in the infarcted heart. We conclude that anti-CD31 immunostaining in Zn-fixed, paraffin-embedded murine tissue offered superior morphology and permitted optimal identification of proliferating endothelial cells during infarct repair.     

Immunohistologic features of minimal-change nephrotic syndrome.

To assess the role of immune mechanisms in the pathogenesis of minimal-change nephrotic syndrome (MCNS), immunohistologic studies were performed on renal biopsy specimens from 31 patients. Glomerular immunoglobulin and/or complement (C3) deposition was present in 20 specimens. Deposits were usually minimal to moderate, granular, and focal in nature. IgM was present in 17 specimens, C3 in 10, IgG in 8 and IgA in only 3. With a mean follow-up of 5 1/2 years, there is no difference in the response to therapy of patients with no glomerular immunoglobulin or C3, those with glomerular immunoglobulin without C3, and those with glomerular C3. These findings indicate that glomerular immune deposits can be seen in a substantial percentage of patients with MCNS, but the minimal and focal nature of the deposits and lack of correlation with response to therapy suggest that immunoglobulin and C3 deposits are nonspecific and have no pathogenetic role.     

Gastrointestinal lymphomas. Immunohistologic study of 23 cases

Twenty-three primary gastrointestinal lymphomas were studied morphologically and immunologically on fresh frozen tissue, and on cell suspension for 16 of them. Polyclonal antibodies reactive with immunoglobulin chains and a panel of 16 monoclonal antibodies reactive with B- and T-cells, histiocytes, and epithelial cells were used. According to the Working Formulation, 5 cases were low grade, 12 intermediate grade, and 5 high grade; 1 case was an extramedullary plasmocytoma. Forty-seven percent were large cell lymphomas and 13% follicular lymphomas. There were 20 (86%) B-cell lymphomas and 2 T-cell lymphomas; one case lacked detectable markers for B-, T-, or histiocytic cells. Monoclonality was demonstrated in 13 out of the 20 B-cell lymphomas, whereas the other 7 expressed pan-B antigens. It is concluded that immunologic studies on frozen surgical material are of precise diagnostic value in gastrointestinal lymphomas, whereas fixed endoscopic biopsies only permit the distinction between lymphomas and undifferentiated carcinomas.     

Immunohistologic heterogeneity of malignant tumors

Intratumor heterogeneity exists in most, if not all, human solid tumors and is reflected by differences in morphology, antigenicity, chromosomal and biochemical markers, invasion, metastatic spread, as well as sensitivity to therapy. Heterogeneity may be the result of genetic and/or environmental mechanisms. To be assessed by immunohistochemistry, heterogeneity of tumor cells has to be expressed by antigenic changes, i.e. quantitative or qualitative differences in antigen expression, secretion or uptake. Consequently, presence and distribution of hormones, steroid hormone receptors (e.g., in mammary carcinomas), blood group antigens (e.g., in diverse carcinomas), and cytoskeletal components, particularly intermediate filament-associated proteins (e.g., in neoplasms of diverse origin), can indicate heterogeneity within a tumor.     

An anatomical heart model with applications to myocardial activation and ventricular mechanics.

A three-dimensional finite element model of the mechanical and electrical behavior of the heart is being developed in a collaboration among Auckland University, New Zealand; the University of California at San Diego, U.S.; and McGill University, Canada. The equations of continuum mechanics from the theory of finite deformation elasticity are formulated in a prolate spheroidal coordinate system and solved using a combination of Galerkin and collocation techniques. The finite element basis functions used for the dependent and independent variables range from linear Lagrange to cubic Hermite, depending on the degree of spatial variation and continuity required for each variable. Orthotropic constitutive equations derived from biaxial testing of myocardial sheets are defined with respect to the microstructural axes of the tissue at the Gaussian quadrature points of the model. In particular, we define the muscle fiber orientation and the newly identified myocardial sheet axis orientation throughout the myocardium using finite element fields with nodal parameters fitted by least-squares to comprehensive measurements of these variables. Electrical activation of the model is achieved by solving the FitzHugh-Nagumo equations with collocation at fixed material points of the anatomical finite element model. Electrical propagation relies on an orthotropic conductivity tensor defined with respect to the local material axes. The mechanical constitutive laws for the Galerkin continuum mechanics model are (1) an orthotropic "pole-zero" law for the passive mechanical properties of myocardium and (2) a Wiener cascade model of the active mechanical properties of the muscle fibers. This chapter concentrates on two aspects of the model: first, grid generation, including both the generation of nodal coordinates for the finite element mesh and the generation of orthotropic material axes at each computational point, and, second, the formulation of constitutive laws suitable for numerically intensive finite element computations. Extensions to this model and applications to the mechanical and electrical function of the heart are described in Chapter 16 by McCulloch and co-workers.     

Immunohistologic diagnosis of avian listeriosis

Listeriosis in a chicken from a small farm was diagnosed by the per-oxidase-antiperoxidase (PAP) method. The animal had diffuse myocarditis and necrotic foci in the liver and in the spleen. PAP technique performed on formalin-fixed tissues confirmed the presumptive diagnosis. Serological and microbiological studies were also done. It is concluded that PAP method is useful for the diagnosis of avian listeriosis allowing retrospective studies with formalin-fixed, paraffin-embedded tissues, avoiding cumbersome microbiological culture.     

An experimental assessment of macroscopic enzyme techniques for the autopsy demonstration of myocardial infarction.

Experimental cardiac infarction produced in dogs by coronary artery ligation has been used to investigate the value of macroscopic histochemical enzyme loss in the diagnosis of myocardial infarction at post-mortem. Creative phosphokinase and non-specific dehydrogenase methods gave the best results but became positive only 5-6 hr after infarction. It is concluded that the method is of limited value in autopsy practice.     

Immunohistologic diagnosis of sarcoidosis. Detection of angiotensin-converting enzyme in sarcoid granulomas

Angiotensin-converting enzyme (ACE) was detected by indirect immunofluorescence examination. ACE was present in the cytoplasm of epithelioid cells in granulomas from 38 of 39 patients who had clinical sarcoidosis but not in any of 37 nonsarcoid granulomas, normal lymph nodes, or splenic controls. Intensity of staining with ACE antiserum was inversely proportional to the degree of fibrosis and was significantly less intense in fibrotic granulomas. The detection of ACE appears to be a distinct aid in establishing a definitive tissue diagnosis of sarcoidosis.     

差异表达蛋白质的分离鉴定及其应用

蛋白质组（proteome）的概念由Wilkins和Williams于1994年首先提出。它源于蛋白质（protein）与基因组（genome）两个词的组合,指一个基因组（genOME）或一个细胞、组织表达的所有蛋白质（PROTein）。蛋白质组学（proteomics）是以蛋白质组为研究对象,阐释所有蛋白质在不同时空的表达谱和功能谱,全景式地揭示生命活动的本质,特别是人体健康与疾病的机制。     

Focal and segmental glomerulosclerosis. Immunohistologic study of 20 renal biopsy specimens

To evaluate the deposition of immunoglobulins and complement and their relationship to sclerotic and nonsclerotic glomerular segments, immunoperoxidase without periodic acid-Schiff counterstain (IMP) and with periodic acid-Schiff counterstain (IMPAS) for IgG, IgA, IgM, and C3 was performed on cryostat-frozen sections using the direct method, along with routine light microscopy and electron microscopy, in a series of 20 renal biopsy specimens from 20 patients with the final diagnosis of focal and segmental glomerulosclerosis. Neither diffuse mesangial nor diffuse glomerular basement membrane deposits were detected by IMP, IMPAS, or electron microscopy. In 18 biopsy specimens, IMPAS demonstrated focal and segmental granular to globular deposits of IgM and/or C3 in sclerotic glomerular segments. In eight biopsy specimens, small granular deposits of IgM and/or C3 deposits were identified in optically normal glomeruli, suggesting that these deposits may precede segmental sclerosis.     

Solid cystic acinar cell tumor of the pancreas. Immunohistologic studies of histogenesis

A case of a very rare solid and cystic tumor of the pancreas in a 25-year-old woman was examined by immunohistochemical methods, e.g. antiglucagon, antisomatostatin, antivip etc. The stainings by the PAP- or biotin-avidin-method were negative and also those with S-100 protein and especially with chromogranin. A tumor of the endocrine system was therefore excluded. Some areas of the tumor showed positive staining with alpha-1-antitrypsin as is known in acinar cell carcinoma. The classification of the neoplasm as an acinar adenoma of the pancreas seems to be well established, mainly because several authors electronmicroscopically demonstrated structures resembling acini and zymogen granules.