Background Nasal polyps in patients with cystic fibrosis (CF) are believed to be phenotypically different than polyps affecting non‐CF patients. The objective of this study was to characterize the expression of inflammatory cytokines within nasal polyps from CF and non‐CF, aspirin‐tolerant patients. “Regulated on activation, normal T‐cell expressed and secreted” (RANTES) is a chemotactic cytokine involved in the recruitment and activation of eosinophils. Multiple molecular studies of non‐CF polyps have established that RANTES may play an important role in nasal polyposis. Our study suggests RANTES may be upregulated in CF polyps relative to non‐CF polyps. Methods Nasal polyps were prospectively obtained from CF and non‐CF, aspirin‐tolerant patients. The Quantibody™ Human Cytokine Array I from RayBiotech, Inc., was used to identify differences in cytokine expression between protein extracts of two polyp groups. Four CF polyp extracts and 4 non‐CF, aspirin‐tolerant polyp extracts were each incubated on identical antibody subarrays, each containing 20 human cytokines in quadruplicate. Western blot analysis confirmed expression of RANTES. Results The protein microarray suggests a greater than 4‐fold upregulation of RANTES in CF polyps relative to non‐CF polyps. Western blot analysis confirmed expression of RANTES in CF polyps. Conclusion Chemokines such as RANTES are responsible for the activation of inflammatory cells within the lamina propria of nasal polyps. We have demonstrated that RANTES may be an important cytokine associated with CF polyps. © 2011 ARS‐AAOA, LLC.
Background: Flow cytometry (FCM) and laser scanning cytometry (LSC) are the routine techniques for fluorescent cell analysis. Recently, we developed a scanning fluorescent microscopy (SFM) technique. This study compares SFM to LSC (two slide-based cytometry, SBC, techniques) and FCM, in experimental and clinical settings. Methods: For the relative cell-frequency determinations, HT29 colorectal cancer cells and Ficoll separated blood mononuclear cells (FSBMCs) were serially diluted (from 1:1 to 1:1,000) and measured by each of the three techniques. For the absolute cell number determinations (only for SBC) FSBMCs were smeared on slides, then HT29 cells were placed on the slide with a micromanipulator (5-50 cells). Tumor cells circulating in the peripheral blood were isolated by magnetic separation from clinical blood samples of colorectal cancer patients. All samples were double-stained by CD45 ECD and CAM5.2 FITC antibodies. For slides,
Automated and quantitative histological analysis can improve diagnostic efficacy in colon sections. Our objective was to develop a parameter set for automated classification of aspecific colitis, ulcerative colitis, and Crohn's disease using digital slides, tissue cytometric parameters, and virtual microscopy. Routinely processed hematoxylin-andeosin-stained histological sections from specimens that showed normal mucosa (24 cases), aspecific colitis (11 cases), ulcerative colitis (25 cases), and Crohn's disease (9 cases) diagnosed by conventional optical microscopy were scanned and digitized in high resolution (0.24 mm/pixel). Thirty-eight cytometric parameters based on morphometry were determined on cells, glands, and superficial epithelium. Fourteen tissue cytometric parameters based on ratios of tissue compartments were counted as well. Leave-one-out discriminant analysis was used for classification of the samples groups. Cellular morphometric features showed no significant differences in these benign colon alterations. However, gland related morphological differences (Gland Shape) for normal mucosa, ulcerative colitis, and aspecific colitis were found (P < 0.01). Eight of the 14 tissue cytometric related parameters showed significant differences (P < 0.01). The most discriminatory parameters were the ratio of cell number in glands and in the whole slide, biopsy/gland surface ratio. These differences resulted in 88% overall accuracy in the classification. Crohn's disease could be discriminated only in 56%. Automated virtual microscopy can be used to classify colon mucosa as normal, ulcerative colitis, and aspecific colitis with reasonable accuracy. Further developments of dedicated parameters are necessary to identify Crohn's disease on digital slides. ' 2008 International Society for Analytical Cytology
The rapidly evolving field of digital microscopy supports the efficient exploitation of inherent information from stained glass slides to offer widespread utilization in breast histopathology. Digital image signals can be accurately measured, integrated into databases and shared through computer networks. Therefore, digital microscopy can boost telepathology-consultation, gradual- and postgradual teaching, proficiency testing, intra- and interlaboratory validation of biomarker screening interpretation, and automated image analysis of biomarker expression for both diagnostics and research applications. This is a brief highlight of the potential of digital microscopy in breast pathology applications.
Automated SFM is an applicable tool for CD4/CD8 ratio determination in peripheral blood samples with QDs.
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