Expression of a Disintegrin and Metalloproteinase 33 Protein in Nasal Polyposis: An Immunohistochemical Study
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A disintegrin and metalloproteinase (ADAM)-33 is a member of matrix metalloproteinases. This protein takes a role in angiogenesis and airway remodeling in asthma. Because histopathological findings of airway remodeling in asthma and nasal polyposis (NP) are similar, the aim of this study was to evaluate the ADAM-33 expression in NP.Immunohistochemical staining of specimens of 47 patients with NP and 8 patients with concha bullosa was performed to detect the expression of ADAM-33. Paraffin blocks were used to identify the expression of ADAM-33 polyclonal antibodies. Immunostaining of epithelial cells, stroma, mesenchymal cells of vessels, and inflammatory cells were analyzed by using light microscopy.Immunopositivity scores in epithelial cells in NP (median, 2; range, 1-3) were significantly higher than those of controls (median, 1.5; range, 1-2; p < 0.001). ADAM-33 staining was increased in the mesenchymal cells of vessels of nasal polyps (median, 2; range, 1-3) compared with control tissues (median, 1.5; range, 1-2; p = 0.006). Although the staining scores of fibroblasts in nasal polyp specimens were also high (median, 3; range 1-3), there was no statistical significance when compared with controls (median 2; range, 1-3; p = 0.228). ADAM-33 immunostaining was not related with the presence of allergies, asthma, and aspirin intolerance (p > 0.05). Moreover, no relationship was found between increased expression of ADAM-33 and the stages of polyp or computerized tomography scores (p > 0.05).This study suggests that the increased expression of ADAM-33 protein may have a role in the pathogenesis of NP.Keywords:
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Nasal Polyps
Our previous work found COX4I2 was associated with angiogenesis in pheochromocytoma. The purpose of this study was to explore the role of COX4I2 in regulating angiogenesis in pheochromocytoma.Distribution of COX4I2 was evaluated by scRNA-seq in one case of pheochromocytoma and the findings were verified by immunostaining. COX4I2 was further knocked down in target cells. Changes of angiogenesis-related genes were evaluated by qPCR in target cells.The scRNA-seq revealed high mRNA expression of COX4I2 in fibroblasts rather than tumor cells. Immunostaining of COX4I2 confirmed its distribution in fibroblasts. Knocking down COX4I2 in NIH3T3 cell line led to significant reduction of angiogenesis-related genes, especially ANG1 and HGF.Fibroblasts mediate the angiogenesis of pheochromocytoma by increasing COX4I2 expression, possibly by affecting ANG1 and HGF.
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AbstractImmunohistochemistry comprises methods used to recognize tissue components as antigens in situ by means of directly or indirectly labeled antibodies, usually (but not always) derived from another species. When applied to cell preparations, the same methods are called immunocytochemistry (Fig. 1), although some authors also use this term for immunostaining of cellular components in tissue sections. Note that compared with immunostaining of vital cells in suspension or cultured monolayers, the sensitivity of immunohistochemical cell-surface staining is considerably reduced because of the decreased amounts of marker antigen represented by the cross-section of the plasma membrane. Also, for certain cellular markers examined in a tissue section, truly peripheral staining may be difficult to distinguish from a rim of cytoplasmic antigen expression. Immunohistochemistry (A) is performed on tissue sections whereas immunocytochemistry (B) is performed on cells in suspensions, smears, or monolayers. Antibodies do not penetrate the surface membrane of living cells; therefore, unequivocal peripheral staining is obtained only for cells in suspension or vital monolayers. With tissue sections or dried (and fixed) cells, it is difficult to distinguish peripheral from cytoplasmic immunostaining unless the antigen is a distinct surface membrane marker. KeywordsCeliac DiseaseMarker AntigenMurine mAbsImmunoenzyme MethodUnwanted InteractionThese keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
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Objective To observe and compare the application between Ventana automatic immunohistochemistry stainer and the manual immunostaining.Methods Several kinds of primary antibodies which were applied to the routine work and the same secondary antibodies were stained by the machine and manual immunohistochemistry operation respectively,and were compared.Results The active results stained by automatic immunohistochemistry stainer of which every index was prior to the manual operation were shown to the well-distributed coloration,exact location,non-edge-effect and clear background.Conclusion The automatic immunohistochemistry stainer is not only high-degree standardizaion and good to normative operation,but also reduces the personal error in manual staining.It could provide exact evidence for clinical pathological diagnosis.
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Tissue samples of 40 patients with histologically confirmed endometrial cancer were analyzed immunohistochemically on paraffin-embedded specimens to detect ras p21 protein expression. The relationship between p21 protein expression and clinicopathological findings was also analyzed. The intensity and distribution of specific cytoplasmatic staining were evaluated semiquantitatively by counting the immunohistochemical H-score. ras p21 expression was found in 30 (75%) of 40 human endometrial carcinomas, regardless of the clinical stage of the disease. Positive immunostaining for p21 was noted in 68% of stage I–II and in all 8 of the advanced stages (III–IV according to FIGO) of endometrial carcinomas. Myometrial invasion was related to ras p21 immunostaining (p = 0.009), however, no correlation between histological findings and ras p21 expression was observed.
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The application of mouse monoclonal antibody for immunostaining the mouse tissues results in a high rate of background noise because of the interaction of the secondary antibody with endogenous immunoglobulins and other immune components. The most advised blocking strategy for the mouse-on-mouse immunostaining is the use of anti-mouse Fab fragments. Nevertheless, the commercial kits containing Fab fragment are costly and unavailable in many research laboratories. In this study, we provide evidence showing the potential of the fluorescent-dye conjugated secondary anti-mouse antibody for reducing the background noise in the mouse-on-mouse immunohistochemistry. Furthermore, our findings demonstrate the inadequacy of goat serum/protein-blocking solution alone as an immunohistochemistry blocking system for reducing the background noise.
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Immunohistochemistry is a technique to examine the expression and distribution of biomolecules in situ using antibodies labeled with enzymes, such as horseradish peroxidase. Generally, both immunohistochemistry and immunofluorescence are termed as immunostaining, because localized biomolecules in situ are detected with antigen-antibody immunoreactions producing color signals. Immunostaining techniques are divided into 3 types: direct, indirect, and amplification methods (peroxidase-antiperoxidase, avidin-biotin complex, and polymer). Because each method has advantages and disadvantages, the method should be selected according to the biological purpose. In this technical note, we describe mechanisms and procedures for immunostaining, including immunohistochemistry and immunofluorescence.
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