ABSTRACT
AIMS: Accurate determination of HER-2 status is critical to identify patients for whom trastuzumab treatment will be of benefit. Although the recommended primary method of evaluation is immunohistochemistry, numerous reports of variability in interpretation have raised uncertainty about the reliability of results. Recent guidelines have suggested that image analysis could be an effective tool for achieving consistent interpretation, and this study aimed to assess whether this technology has potential as a diagnostic support tool. METHODS AND RESULTS: Across a cohort of 275 cases, image analysis could accurately classify HER-2 status, with 91% agreement between computer-aided classification and the pathology review. Assessment of the continuity of membranous immunoreactivity in addition to intensity of reactivity was critical to distinguish between negative and equivocal cases and enabled image analysis to report a lower referral rate of cases for confirmatory fluorescence in situ hybridization (FISH) testing. An excellent concordance rate of 95% was observed between FISH and the automated review across 136 informative cases. CONCLUSIONS: This study has validated that image analysis can robustly and accurately evaluate HER-2 status in immunohistochemically stained tissue. Based on these findings, image analysis has great potential as a diagnostic support tool for pathologists and biomedical scientists, and may significantly improve the standardization of HER-2 testing by providing a quantitative reference method for interpretation.
Subject(s)
Breast Neoplasms/chemistry , Breast Neoplasms/diagnosis , Image Processing, Computer-Assisted/methods , Receptor, ErbB-2/analysis , Algorithms , Antibodies, Monoclonal/therapeutic use , Antibodies, Monoclonal, Humanized , Antineoplastic Agents/therapeutic use , Biomarkers, Tumor/analysis , Breast Neoplasms/drug therapy , Breast Neoplasms/genetics , Cohort Studies , Diagnosis, Computer-Assisted , Female , Genes, erbB-2 , Humans , Image Processing, Computer-Assisted/standards , Immunohistochemistry/methods , In Situ Hybridization, Fluorescence , TrastuzumabABSTRACT
Supplementation of the divalent cations calcium and magnesium to submerged cultures of Streptomyces hygroscopicus var. geldanus greatly influenced morphological development and secondary metabolite synthesis. The disparate response could be explained in terms of the differential effects of Ca2+ and Mg2+ ions on cell surface hydrophobicity. Cultures supplemented with calcium ions were found to be hydrophobic, which resulted in cell concentration-dependent aggregation. In contrast, those grown in a magnesium-rich medium were found to be hydrophilic with the organism growing as freely dispersed filaments that synthesised geldanamycin at an optimal rate in comparison to hydrophobic pellets.
Subject(s)
Calcium/metabolism , Magnesium/metabolism , Streptomyces/cytology , Streptomyces/metabolism , Benzoquinones/metabolism , Hydrophobic and Hydrophilic Interactions , Lactams, Macrocyclic/metabolism , Streptomyces/chemistry , Streptomyces/growth & developmentABSTRACT
Tissue Microarrays facilitate high-throughput immuohistochemistry; however, there are key bottlenecks apparent in their analysis, particularly when conducting microscope-based manual reviews. Traditionally Tissue Microarray assessments were performed using a microscope where results were either transcribed or dictated and subsequently entered into flat-file spreadsheets. This process is labour intensive, prone to error and negates the advantages of the high-throughput Tissue Microarray format. In addition, human interpretations of staining intensity parameters are highly subjective and therefore prone to inter- and intra-observer variability. The advent of Virtual Slides has permitted the review of tissue slides across the Internet. In addition, this new technology enables the creation of software solutions to assist in the manual and automated review of Tissue Microarrays, through the use of computer aided image analysis. There are numerous academically developed and commercially available applications which assist in Tissue Microarray reviews; functionality of these systems range in complexity and application domains. The review which follows describes these systems and outlines technical considerations to be assessed when deciding on a Tissue Microarray workflow solution.
Subject(s)
Microscopy/methods , Proteins/analysis , Tissue Array Analysis/methods , Humans , Immunohistochemistry , SoftwareABSTRACT
The diverse morphology of the filamentous organism Streptomyces hygroscopicus var. geldanus was characterised by image analysis under various environmental conditions. In the presence of surfactant compounds, a significant decrease in the mean pellet diameter was observed. Cell aggregation was also influenced by spore inoculum level, with high concentrations reducing pellet size. In addition, the dispersion of pellets was found to increase with the inclusion of glass beads to submerged shake-flask cultures. In all cases, production of the secondary metabolite geldanamycin was determined to be dependent on the morphological profile of the organism, with a concomitant increase of 88% in geldanamycin yield observed as the mean pellet diameter was reduced by 70%. Thus, to maximise the yield of geldanamycin, it is necessary to limit pellet formation in Streptomyces hygroscopicus var. geldanus to an appropriate size.
