Immunohistochemical Characterization (immunohistochemical + characterization)

Distribution by Scientific Domains

Selected Abstracts

Immunohistochemical Characterization of Hepatic Malondialdehyde and 4-Hydroxynonenal Modified Proteins During Early Stages of Ethanol-Induced Liver Injury

ALCOHOLISM, Issue 6 2003
Brante P. Sampey
Background: Chronic ethanol consumption is associated with hepatic lipid peroxidation and the deposition or retention of aldehyde-adducted proteins postulated to be involved in alcohol-induced liver injury. The purpose of this study was to characterize hepatocellular formation of aldehyde-protein adducts during early stages of alcohol-induced liver injury. Methods: Female Sprague Dawley® rats were subjected to the intragastric administration of a low-carbohydrate/high-fat total enteral nutrition diet or a total enteral nutrition diet containing ethanol for a period of 36 days. Indexes of hepatic responses to ethanol were evaluated in terms of changes in plasma alanine aminotransferase activity, hepatic histopathologic analysis, and induction of cytochrome P-4502E1 (CYP2E1). Immunohistochemical methods were used to detect hepatic proteins modified with malondialdehyde (MDA) or 4-hydroxynonenal (4-HNE) for subsequent quantitative image analysis. Results: After 36 days of treatment, rats receiving the alcohol-containing diet displayed hepatic histopathologies characterized by marked micro- and macrosteatosis associated with only minor inflammation and necrosis. Alcohol administration resulted in a 3-fold elevation of plasma alanine aminotransferase activity and 3-fold increases (p < 0.01) in hepatic CYP2E1 apoprotein and activity. Quantitative immunohistochemical analysis revealed significant (p < 0.01) 5-fold increases in MDA- and 4-HNE modified proteins in liver sections prepared from rats treated with alcohol. The MDA- or 4-HNE modified proteins were contained in hepatocytes displaying intact morphology and were colocalized primarily with microvesicular deposits of lipid. Aldehyde-modified proteins were not prevalent in parenchymal or nonparenchymal cells associated with foci of necrosis or inflammation. Conclusions: These results suggest that alcohol-induced lipid peroxidation is an early event during alcohol-mediated liver injury and may be a sensitizing event resulting in the production of bioactive aldehydes that have the potential to initiate or propagate ensuing proinflammatory or profibrogenic cellular events. [source]

Immunohistochemical Characterization of Neurones in the Hypoglossal Nucleus of the Pig

W. Sienkiewicz
Summary With 4 figures and 1 table In this study, the presence of several neurotransmitters and transmitter synthesizing enzymes was studied in hypoglossal nucleus (HN) of the juvenile (4 months old) female pigs (n = 3). Double-labeling immunofluorescence revealed neurones expressing cholinacetyltranspherase (ChAT), calcitonin gene-related peptide (CGRP), nitric oxide synthase (NOS), and somatostatin (SOM). Nerve fibers within HN were ChAT, CGRP, NOS, SOM, substance P (SP), Leu-5-enkephalin (Leu-5-Enk), ß-dopamine hydroxylase (DßH), neuropeptide Y (NPY) positive. Virtually all the perikarya contained ChAT, whereas CGRP was present in 47% of the neurones. Nerve cell bodies containing NOS or SOM were only occasionally observed. Immunoreactive nerve fibers were found in a close vicinity of the perikarya, often forming baskets around nerve cell bodies. The results obtained were compared with similar data obtained in other species. The presence of immunoreactive structures, origin of the nerve fibers, and functional significance of the findings are discussed. [source]

Immunohistochemical characterization of glomerular inflammatory cells and expression of adhesion molecules in anti-glomerular basement membrane (anti-GBM) glomerulonephritis induced in WKY rats with monoclonal anti-GBM antibodies of different subclasses

Tadashi Kado
According to previous report, adhesion of CD8-positive cells and macrophages to glomerular endotherial cells through the lymphocyte function-associated antigen-1 (LFA-1)/intercellular adhesion molecule-1 (ICAM-1) pathway is crucial for the initiation and subsequent progression of anti-glomerular basement membrane (anti-GBM) antibody-induced glomerulonephritis (anti-GBM nephritis) in WKY rats. In the present study glomerular inflammatory cell infiltration and LFA-1/ICAM-1 expression were examined in anti-GBM nephritis induced in WKY rats with monoclonal anti-GBM antibodies of different subclasses: IgG1, IgG2a, and IgG2b. The IgG2a and IgG2b subclasses induced significant proteinuria from day 3 as compared with the IgG1 subclass. Glomerular infiltration of macrophages and CD8-positive cells after administration of IgG2a and IgG2b subclass antibodies was significantly elevated compared to that for the IgG1 subclass. The intensity of glomerular ICAM-1 immunostaining by the IgG2a and IgG2b subclass antibodies tended to be stronger than that by the IgG1 subclass. Glomerular LFA-1-positive cell infiltration by the IgG2a and IgG2b subclasses was significantly higher than that of the IgG1 subclass. These results demonstrate that monoclonal antibodies belonging to the IgG2a and IgG2b subclasses strongly induce glomerular infiltration of inflammatory cells and expression of adhesion molecules in rat anti-GBM nephritis. [source]

Clinical and histologic evidence of salivary gland restoration supports the efficacy of rituximab treatment in Sjögren's syndrome,

J. Pijpe
Objective To assess the effect of rituximab (anti-CD20 antibody) therapy on the (immuno)histopathology of parotid tissue in patients with primary Sjögren's syndrome (SS) and the correlation of histologic findings with the flow rate and composition of parotid saliva. Methods In a phase II study, an incisional parotid biopsy specimen was obtained from 5 patients with primary SS before and 12 weeks after rituximab treatment (4 infusions of 375 mg/m2). The relative amount of parotid parenchyma, lymphocytic infiltrate, and fat, and the presence/quantity of germinal centers and lymphoepithelial duct lesions were evaluated. Immunohistochemical characterization was performed to analyze the B:T cell ratio of the lymphocytic infiltrate (CD20, CD79a, CD3) and cellular proliferation in the acinar parenchyma (by double immunohistologic labeling for cytokeratin 14 and Ki-67). Histologic data were assessed for correlations with the parotid flow rate and saliva composition. Results Four patients showed an increased salivary flow rate and normalization of the initially increased salivary sodium concentration. Following rituximab treatment, the lymphocytic infiltrate was reduced, with a decreased B:T cell ratio and (partial) disappearance of germinal centers. The amount and extent of lymphoepithelial lesions decreased in 3 patients and was completely absent in 2 patients. The initially increased proliferation of acinar parenchyma in response to inflammation was reduced in all patients. Conclusion Sequential parotid biopsy specimens obtained from patients with primary SS before and after rituximab treatment demonstrated histopathologic evidence of reduced glandular inflammation and redifferentiation of lymphoepithelial duct lesions to regular striated ducts as a putative morphologic correlate of increased parotid flow and normalization of the salivary sodium content. These histopathologic findings in a few patients underline the efficacy of B cell depletion and indicate the potential for glandular restoration in SS. [source]

Oncocytic lesions of the ophthalmic region

Purpose To make a nationwide clinicopathologic study of oncocytic lesions in the ophthalmic region and to characterize their cytokeratin(CK) expression. Methods All histologically diagnosed oncocytic lesions in the ophthalmic region registered in Denmark over a 25-year period were collected and re-evaluated using a monoclonal antimitochondrial antibody (MU213-UC). Clinical data were registered. Immunohistochemical characterization was performed with a panel of anticytokeratin antibodies. Results A total of 34 oncocytic lesions were identified. The incidence that required surgical intervention in the Danish population could be approximated to 0.3 lesions per million capita per year. The age of the patients ranged from 45 to 89 years with a peak incidence in the 8th decade. Female gender was twice as common as male. Lesions were typically described as red-brown, cystic and slow growing. The antimitochondrial antibody MU213-UC produced a distinct and intense immunostaining. 26 of the lesions originated from the caruncle, three in the conjunctiva, two in the lacrimal sac, one at the semilunar plica, one on the eyelid margin and one peripunctal. Lesions were histologically classified as adenoma(oncocytoma) (26), hyperplasia (4) and metaplasia (4). Basal-type oncocytic cells reacted with antibodies against CK5/6, CK7, CK8, CK13, CK14, CK 17, CK18, CK19 and suprabasal cells with CK4, CK7, CK8, CK18 and CK19. Antibodies against CK1+10 and CK20 showed no reaction. Conclusion Oncocytic lesions of the ophthalmic region most frequently present as a caruncular oncocytoma. The cytokeratin profile is similar to the lacrimal - and accessory lacrimal gland duct elements and supports the theory that these lesions originate from lacrimal , and accessory lacrimal glands. [source]

Immunohistochemical characterization of guided bone regeneration at a dehiscence-type defect using different barrier membranes: an experimental study in dogs

Frank Schwarz
Abstract Objectives: The aim of the present study was to evaluate immunohistochemically the pattern of guided bone regeneration (GBR) using different types of barrier membranes. Material and methods: Standardized buccal dehiscence defects were surgically created following implant bed preparation in 12 beagle dogs. Defects were randomly assigned to six different GBR procedures: a collagen-coated bone grafting material (BOC) in combination with either a native, three cross-linked, a titanium-reinforced collagen membrane, or expanded polytetrafluorethylene (ePTFE), or BOC alone. After 1, 2, 4, 6, 9, and 12 weeks of submerged healing, dissected blocks were processed for immunohistochemical (osteocalcin , OC, transglutaminase II , angiogenesis) and histomorphometrical analysis [e.g., bone-to-implant contact (BIC), area of new bone fill (BF)]. Results: In general, angiogenesis, OC antigen reactivity, and new bone formation mainly arose from open bone marrow spaces at the bottom of the defect and invaded the dehiscence areas along the implant surface and BOC. At 4 weeks, membranes supporting an early transmembraneous angiogenesis also exhibited some localized peripheral areas of new bone formation. However, significantly increasing BIC and BF values over time were observed in all groups. Membrane exposure after 10,12 weeks was associated with a loss of the supporting alveolar bone in the ePTFE group. Conclusion: Within the limits of the present study, it was concluded that (i) angiogenesis plays a crucial role in GBR and (ii) all membranes investigated supported bone regeneration on an equivalent level. [source]

Alternatively activated macrophages (M2 macrophages) in the skin of patient with localized scleroderma

Nobuyo Higashi-Kuwata
Abstract:, Localized scleroderma is a connective tissue disorder that is limited to the skin and subcutaneous tissue. Macrophages have been reported to be particularly activated in patients with skin disease including systemic sclerosis and are potentially important sources for fibrosis-inducing cytokines, such as transforming growth factor ,. To clarify the features of immunohistochemical characterization of the immune cell infiltrates in localized scleroderma focusing on macrophages, skin biopsy specimens were analysed by immunohistochemistry. The number of cells stained with monoclonal antibodies, CD68, CD163 and CD204, was calculated. An evident macrophage infiltrate and increased number of alternatively activated macrophages (M2 macrophages) in their fibrotic areas were observed along with their severity of inflammation. This study revealed that alternatively activated macrophages (M2 macrophages) may be a potential source of fibrosis-inducing cytokines in localized scleroderma, and may play a crucial role in the pathogenesis of localized scleroderma. [source]

Characterization of Zebrafish Cx43.4 Connexin and its Channels

T. Desplantez
Connexins (Cx) form intercellular junctional channels which are responsible for metabolic and electrical coupling. We report here on the biochemical and immunohistochemical characterization of zebrafish connexin zfCx43.4, an orthologue of mammalian and avian Cx45, and the electrophysiological properties of junctional channels formed by this protein. The investigations were performed on transfected COS-7 cells or HeLa cells. Using site-directed antibodies, zfCx43.4 cDNA (GenBank accession no. X96712) was demonstrated to code for a protein with a Mr of 45 000. In transfected cells, zfCx43.4 was localized in cell-cell contact areas as expected for a gap junction protein. zfCx43.4 channels were shown to transfer Lucifer Yellow. The multichannel currents were sensitive to the transjunctional voltage (Vj). Their properties were consistent with a two-state model and yielded the following Boltzmann parameters for negative/positive Vj: Vj,0= -38.4/41.9 mV; gj,min= 0.19/0.18; z = 2.6/2.3. These parameters deviate somewhat from those of zfCx43.4 channels expressed in Xenopus oocytes and from those of Cx45, an orthologue of zfCx43.4, expressed in mammalian cells or Xenopus oocytes. Conceivably, the subtle differences may reflect differences in experimental methods and/or in the expression system. The single channel currents yielded two prominent levels attributable to a main conductance state (,j,main= 33.2 ± 1.5 pS) and a residual conductance state (,j,residual= 11.9 ± 0.6 pS). [source]

Lobular capillary hemangioma of the oral mucosa: Clinicopathological study of 43 cases with a special reference to immunohistochemical characterization of the vascular elements

Makoto Toida
Clinical and histopathological features were investigated in 43 cases of oral lobular capillary hemangiomas (LCH) with a special reference to characteristics of the vascular elements. The lesions affected females more than males by a ratio of 1:1.5. Average age of the patients was 52.7 years. The lesions involved the gingiva (n = 15), the tongue (n = 13), the labial mucosa (n = 10) and other sites. The lesions appeared usually as a pedunculated mass with ulceration; size of the lesions was up to 15 mm. Histologically, a lobular area and an ulcerative area were distinguished. The density of vessels was about 1045/mm2 and 160/mm2 in the lobular and ulcerative areas, respectively. The average diameter of the vascular lumen was 9.1 5.6 mm (range: 2.8,42.0 mm) and 18.8 20.9 mm (range: 5.6,139.7 mm) in the lobular and ulcerative areas, respectively. In the lobular area, most of the vessels had an inner layer of endothelial cells showing positive reaction for von Willebrand factor (vWF) and CD34, as well as an outer layer of mesenchymal cells showing positive reaction for alpha-smooth muscle actin (ASMA). However, in the ulcerative area, there was a variety of types of vessels consisting of various proportions of both endothelial and ASMA-positive perivascular mesenchymal cells. These results indicate that most of the vascular elements in the lobular area resemble more pericapillary microvascular segments than they do capillaries. Thus, the authors propose the term ,lobular pericapillary hemangioma' to represent this type of lesion. [source]

Correlated morphological and chemical phenotyping in myenteric type V neurons of porcine ileum

Axel Brehmer
Abstract The study was aimed at the immunohistochemical characterization of myenteric Stach type V neurons of the pig ileum that were not included in the widely used Dogiel classification. So far, this conspicuous population has been defined morphologically on the basis of silver-impregnated specimens only. By using neurofilament immunohistochemistry, type V neurons that occur singly or in aggregates could be identified unequivocally and could be distinguished from other smoothly contoured myenteric neurons, i.e., type II and type IV. Double-labeling immunohistochemistry revealed a number of potentially neuroactive substances or their synthesizing enzymes to be present in type V neurons. Choline acetyltransferase immunoreactivity (-ir) was found in all type V neurons, whereas neuronal nitric oxide synthase was detected in none. Leu-enkephalin-ir was found within 92.3%, somatostatin (SOM)-ir within 91.1%, calcitonin gene-related peptide (CGRP)-ir within 80.6% and met-enkephalin-ir within 74.7% of type V neurons. Triple-labeling immunohistochemistry was applied to address the question of a specific chemical coding for myenteric type V neurons. In contrast to other combinations of neuroactive substances/enzymes that were found in both type V and other, nontype V neurons, SOM/CGRP-ir was the only combination observed exclusively within type V neurons. Both substances were colocalized in 79.3% of type V neurons. This colocalization discriminates four-fifths of the type V neurons chemically from both type II neurons (CGRP positive, SOM negative) and type IV neurons (CGRP negative, SOM positive), which both share, at first glance, a similar morphology with type V neurons. These results further support the concept of a close correlation between morphologically defined neuronal type and chemical coding and, it is likely, also function in the enteric nervous system of larger mammals. J. Comp. Neurol. 453:1,9, 2002. © 2002 Wiley-Liss, Inc. [source]

Histopathological and immunohistochemical characterization of canine prostate cancer

THE PROSTATE, Issue 5 2008
Chen-Li Lai
Abstract Background In this study we try to identify the origin of canine prostate cancer (cPC) by classifying the tumors histological subtypes and relate these subtypes to their combined expressional characteristics of several tissue specific and differentiation markers. Methods cPCs were examined histomorphologically and by immunohistochemical detection of the cytokeratin markers CK14, HMWCK, CK5, CK18, and CK7, and of the markers UPIII, PSA and PSMA. Results Histopathologically, six growth patterns could be differentiated. The most frequent patterns were solid, cribriform and micropapillary growth patterns, while sarcomatoid, small acinar/ductal, and tubulo-papillary growth patterns were less frequent present. Solid growth patterns were significantly (P,=,0.027) more often seen in castrated dogs. Immunohistochemically, about half of the cPC cases showed expression of PSA (8/20) and PSMA (10/20); 85% and 60% of the cPC expressed UPIII (17/20) and CK7 (12/20), while 13 and 12 cPC expressed CK5 and CK14, respectively; all cPC expressed CK18. CK14 was significantly more often and UPIII less frequent expressed in the solid growth patterns than in the micropapillary and cribriform patterns, respectively. Conclusions Canine prostate cancer appear to be more aggressive and of a less differentiated type than most common human prostate cancers. Comparing the expression patterns of the markers in cPC to those in normal canine prostate tissue, cPC most likely originates from the collecting ducts rather than from the peripheral acini. Given also the fact that canine prostate cancer is unresponsive to androgen withdrawal therapy, canine prostate cancer mostly resembles human, androgen refractory, poorly differentiated prostate cancer. Prostate 68: 477,488, 2008. © 2008 Wiley-Liss, Inc. [source]

Interstitial cells in the human prostate: A new therapeutic target?

THE PROSTATE, Issue 4 2003
Frank Van der Aa
Abstract BACKGROUND Interstitial cells have been described in different human organs, including gut and bladder. In the gut they function as pacemaker cells, generating slow wave potentials. Absence or defects in these cells result in motility disorders. In the bladder these cells express the vanilloid receptor and may contribute to the working mechanism of vanilloid therapy. Recently, slow wave potentials and interstitial cells were described in the guinea-pig prostate. In this study we describe the presence of interstitial cells in the human prostate gland. METHODS We performed immunohistochemical staining for c-kit, vanilloid receptor (VR1), cannabinoid receptor (CB1) connexin43, and neurofilament on fresh frozen tissue from 14 prostatectomy specimens. RESULTS A large number of cells with a stellate aspect were noticed under the basal layer of the prostatic duct system and in between the smooth muscle cells. They were immunoreactive for c-kit, VR1, and connexin43 but not to CB1 or neurofilament. CONCLUSIONS There is evidence for interstitial cells in the human prostate. Taken together their topography and immunohistochemical characterization, the discovery of slow wave potentials in guinea pig prostate and the knowledge of interstitial cells in other organs, interstitial cells are likely to be involved in normal prostate physiology. Prostate 56: 250,255, 2003. © 2003 Wiley-Liss, Inc. [source]

Morphological and immunohistochemical characterization of paraneoplastic cerebellar degeneration associated with Yo antibodies

A. Storstein
Introduction,,, Immunohistochemical studies of paraneoplastic cerebellar degeneration (PCD) are rare, and the findings vary. Materials and methods,,, We performed morphological and immunohistochemical characterization of the brain, medulla and tumour of two patients with PCD, Yo antibodies and ovarian adenocarcinoma. Results,,, The cerebellum of both patients had extensive loss of Purkinje cells. Microglia activation and T cells were found in the cerebellum, but B cells or deposits of IgG or complement were not detected. Microglia activation was also present in the brain stem and medulla. T cells were found in the ovarian adenocarcinoma. Conclusion,,, PCD is characterized by loss of Purkinje cells and microglia activation, and the presence of T cells indicates cellular immune reactions in PCD and in ovarian cancer. [source]

Identification of Muir,Torre syndrome among patients with sebaceous tumors and keratoacanthomas

CANCER, Issue 5 2005
Role of clinical features, immunohistochemistry, microsatellite instability
Abstract BACKGROUND The Muir,Torre syndrome (MTS) is an autosomal-dominant genodermatosis characterized by the presence of sebaceous gland tumors, with or without keratoacanthomas, associated with visceral malignancies. A subset of patients with MTS is considered a variant of the hereditary nonpolyposis colorectal carcinoma, which is caused by mutations in mismatch-repair genes. The objective of the current study was to evaluate whether a combined clinical, immunohistochemical, and biomolecular approach could be useful for the identification of Muir,Torre syndrome among patients with a diagnosis of sebaceous tumors and keratoacanthomas. METHODS The authors collected sebaceous skin lesions and keratoacanthomas recorded in the files of the Pathology Department of the University of Modena during the period 1986,2000. Through interviews and examination of clinical charts, family trees were drawn for 120 patients who were affected by these skin lesions. RESULTS Seven patients also were affected by gastrointestinal tumors, thus meeting the clinical criteria for the diagnosis of MTS. In the MTS families, a wide phenotypic variability was evident, both in the spectrum of visceral tumors and in the type of skin lesions. Microsatellite instability was found in five MTS patients: These patients showed concordance with immunohistochemical analysis; moreover, a constitutional mutation in the MSH2 gene was found in 1 patient. Lack of expression of MSH2/MSH6 or MLH1 proteins was evident in the skin lesions and in the associated internal malignancies of 3 patients and 2 patients with MTS, respectively. CONCLUSIONS The clinical, biomolecular, and immunohistochemical characterization of sebaceous skin lesions and keratoacanthomas may be used as screening for the identification of families at risk of MTS, a disease that is difficult to recognize and diagnose. Cancer 2005. © 2005 American Cancer Society. [source]