摘要:

Immunohistochemical evaluation of hormone receptors for breast cancer has been performed parallel to biochemical assays. Recently, immunohistochemistry has tended to substitute the biochemical method in Japan. To clarify the factors concerned and problems to be resolved, we reviewed our evaluation system for hormone receptors by immunohistochemistry from 1990. A total of 861 breast cancer samples were examined by immunohistochemistry and biochemistry. In 3 main periods, phase 1 (1990–1993), phase 2 (1995–1998), and phase 3 (1999–2001), increasing sensitivity of the immunohistochemical method was provided by commercially available staining systems and shown to range from 83.6% (phase 1) to 92.0% (phase 3). The highly sensitive procedures of the antigen retrieval and peroxidase-conjugated polymer method are main contributing factors. The authors examined how these procedures influenced the distribution of positive cell population; concordance rate, including sensitivity and specificity; cutoff points; and evaluation categories. The correlation between biochemistry and immunohistochemistry was extensively studied in the 1980s and 1990s. In reference to the progress achieved in the United States and United Kingdom to control the current situation in Japan, it should be recognized that recently developed, highly sensitive procedures boost the immunoreactivity, which will affect the basic factors for technical validation.

ISSN:1541-2016    

出版商:OVID    

年代:2003

数据来源: OVID

摘要:

The immunohistochemical detection of the mismatch repair (MMR) proteins is used as a screening test with microsatellite instability for the detection of hereditary nonpolyposis colon cancer (HNPCC). The authors describe a simple and cost-effective method using a pressure cooker and microwave oven for antigen retrieval and a modified method for applying a commercial biotin blocking kit. Colorectal tumors of 20 patients of the HNPCC spectrum were included in this study. Eighty paraffin sections were cut and submitted for immunohistochemical analysis using a routine protocol and a pressure cooker protocol. Parallel sections for biotin blocking were also run, including the modified biotin block for each protocol. The sections were incubated with the following antibodies: MLH1, MSH2, MSH6, and PMS2. All cases examined exhibited a normal expression of the MMR proteins in the nucleus and adjacent nonneoplastic tissue elements and consequently defined as having a normal expression of these proteins. Cases with tumor that exhibited a loss of the nuclear staining with the MMR proteins with a concurrent staining of the adjacent nonneoplastic cells were classified as abnormal MMR expression. The series of 20 cases using pressure cooker antigen retrieval produced superior results to the routine immunohistochemical protocol used previously in our laboratory. The modified biotin block also gave consistent results. The reproducibility and consistency of this procedure has resulted it in being used routinely for suspected HNPCC cases, both current and archival.

ISSN:1541-2016    

出版商:OVID    

年代:2003

数据来源: OVID

摘要:

Although matrix metalloproteinases (MMPs) are known to be involved in the development of atherosclerosis and the instability of atheromatous plaques, much remains to be learned about their roles at the tissue level. To help clarify this area, we established a new double staining method using film in situ zymography and immunohistochemistry. Using this technique, a comprehensive analysis of the gelatinolytic activity in human vessel tissue demonstrated that gelatinolytic activity is enhanced in the shoulder region and fibrous cap at superficial areas of the atheromatous plaque in the presence of thrombolysis. Enzyme assay clarified high activity in the superficial area (7.50 ± 5.04 U/mg weight;P< 0.001). Gelatin zymography also indicated that addition of the antiplatelet agent, trapidil, alters the amount of secretion of MMP-2 and MMP-9 and their activation ratio. This novel approach to detect the activity of gelatinases in resected tissues may aid in the selection of optimal treatment of individual patients.

ISSN:1541-2016    

出版商:OVID    

年代:2003

数据来源: OVID

摘要:

We tested a recently developed flexible method of separation and quantification of immunohistochemical staining by means of color image analysis. An algorithm was recently developed to deconvolve the color information acquired with RGB cameras, to calculate the contribution of each of the applied stains, based on the stain-specific RGB absorption. The algorithm was tested using a set of lung-tumor samples labeled for the detection of Ki-67, an antigen expressed in proliferating cells, covering a wide range of staining levels. Quantification of the labeling was compared with HSI-based segmentation and manual analysis of the same samples. The recently developed deconvolution method performed significantly better than the HSI based system when compared to manual counting as gold standard. The deconvolution system showed significantly reduced variability in the LI determination, especially of highly labeled control samples. This resulted in significant increase in sensitivity of classification of samples with increased KI-67 labeling without changing the specificity, when compared to the HSI based method.

ISSN:1541-2016    

出版商:OVID    

年代:2003

数据来源: OVID

摘要:

B5 clearing is a step used before immunoperoxidase staining to remove the precipitated mercuric chloride deposits caused by B5 fixation of tissue. In the B5 clearing procedure, the slides are treated with Lugol's iodine and 5% sodium thiosulfate before antigen retrieval and the application of the primary antibody. The goal of this project was to study the effect of the B5 clearing protocol on immunoperoxidase staining on paraffin-embedded tissue, which has not been previously reported in a series of antibodies. We evaluated 75 antibodies using the 2-step clearing protocol and performed paired immunoperoxidase staining on the Ventana ES instrument, with and without the clearing protocol. We found that among 75 antibodies studied, 3 (CD5, CD30, and synaptophysin) showed total obliteration of reactivity, and 3 (ALK, Ulex, and GFAP) showed partial reduction of the staining compared with the controls. Pathologists must be aware of the possible false-negative staining effect caused by the routinely used B5 clearing protocol. Control tissues must receive the same clearing protocol (i.e., placed on case slides) to ensure detection of this effect.

ISSN:1541-2016    

出版商:OVID    

年代:2003

数据来源: OVID

摘要:

A 63-year-old man was admitted to our hospital for complaints of a painless knot in his right testicle. The patient underwent orchiectomy for suspicion of malignancy. Pathologic examination detected a firm, circumscribed mass that consisted histologically of noncohesive, large, undifferentiated tumor cells diffusely infiltrating the testicular interstice. Because of the patient's advanced age and the structure of the neoplastic cells, the differential diagnosis favored a lymphoma over a malignant germ cell tumor but also included a dedifferentiated metastatic neoplasm. Immunohistochemistry was tailored accordingly and was completed using three successive panels of antibodies. Immunostaining with the first panel of antibodies directed against leukocyte epitopes (CD45, CD20, and CD3) remained negative and made a lymphoma unlikely. The subsequent panel (cytokeratin AE1/3, cytokeratin 18, and HMB-45) helped to rule out a malignant melanoma and aided to settle the diagnosis of a metastatic carcinoma. The reaction pattern of the last panel of antibodies pointed to a pulmonary origin of the putative primary tumor (PSA-negative, TTF-1-positive, and CK20-negative). The diagnosis of a metastasized poorly differentiated adenocarcinoma of the lung was confirmed by autopsy 5 months later. This case represents the extremely rare occurrence of a testicular metastasis as a primary manifestation of an occult neoplasm and shows the usefulness of an integrated site-specific clinicomorphologic approach that should precede and guide the choice of diagnostic immunoreagents.

ISSN:1541-2016    

出版商:OVID    

年代:2003

数据来源: OVID

ISSN:1541-2016    

出版商:OVID    

年代:2003

数据来源: OVID