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      Evaluation of Mitotic Activity Index in Breast Cancer Using Whole Slide Digital Images

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          Abstract

          Introduction

          Mitotic Activity Index (MAI) is an important independent prognostic factor and an integral part of the breast cancer grading system. Thus, correct estimation of this prognostically relevant feature is essential for guiding treatment decision and assessing patient prognosis.

          The aim of this study was to validate the use of high resolution Whole Slide Images (WSI) in estimating MAI in breast cancer specimens.

          Methods

          MAI was evaluated in 100 consecutive breast cancer specimens by three observers on two occasions, microscopically and on WSI with a wash out period of 4 months. MAI was also translated to mitotic scores as in grading. Inter- and intra-observer agreement between microscopic and digital MAI counts and scores was measured.

          Results

          Almost perfect inter-observer agreements were obtained from counting MAI using a conventional microscope (intra-class correlation coefficient (ICCC) 0.879) as well as on WSI (ICCC 0.924). K coefficients reflected good inter-observer agreements among observers' microscopic mitotic scores (average kappa 0.642). Comparable results were also observed among digital mitotic scores (average kappa 0.635). There was strong to perfect intra-observer agreements between MAI counts and mitotic scores for the two diagnostic modalities (ICCC 0.716–0.863, kappa 0.506–0.617). There were no significant differences in mitotic scores using both diagnostic modalities.

          Conclusion

          Scoring mitoses using WSI in breast cancer seems to be just as reliable and reproducible as when using a microscope. Further development of software and image quality will definitely encourage the use of WSI in routine pathology practice.

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          Most cited references27

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          Prognostic factors in breast cancer. College of American Pathologists Consensus Statement 1999.

          Under the auspices of the College of American Pathologists, a multidisciplinary group of clinicians, pathologists, and statisticians considered prognostic and predictive factors in breast cancer and stratified them into categories reflecting the strength of published evidence. Factors were ranked according to previously established College of American Pathologists categorical rankings: category I, factors proven to be of prognostic import and useful in clinical patient management; category II, factors that had been extensively studied biologically and clinically, but whose import remains to be validated in statistically robust studies; and category III, all other factors not sufficiently studied to demonstrate their prognostic value. Factors in categories I and II were considered with respect to variations in methods of analysis, interpretation of findings, reporting of data, and statistical evaluation. For each factor, detailed recommendations for improvement were made. Recommendations were based on the following aims: (1) increasing uniformity and completeness of pathologic evaluation of tumor specimens, (2) enhancing the quality of data collected about existing prognostic factors, and (3) improving patient care. Factors ranked in category I included TNM staging information, histologic grade, histologic type, mitotic figure counts, and hormone receptor status. Category II factors included c-erbB-2 (Her2-neu), proliferation markers, lymphatic and vascular channel invasion, and p53. Factors in category III included DNA ploidy analysis, microvessel density, epidermal growth factor receptor, transforming growth factor-alpha, bcl-2, pS2, and cathepsin D. This report constitutes a detailed outline of the findings and recommendations of the consensus conference group, organized according to structural guidelines as defined.
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            Breast carcinoma malignancy grading by Bloom-Richardson system vs proliferation index: reproducibility of grade and advantages of proliferation index.

            Questions of reproducibility and efficacy of histologic malignancy grading relative to alternative proliferation index measurements for outcome prediction remain unanswered. Microsections of specimens from the Cooperative Breast Cancer Tissue Resource (CBCTR) were evaluated by seven pathologists for reproducibility of grade and classification. Nuclear figure classification was assessed using photographs. Grade was assigned by the Bloom-Richardson method, Nottingham modification. Proliferation index was evaluated prospectively by deoxyribose nucleic acid precursor uptake with thymidine (autoradiographic) or bromodeoxyuridine (immunohistochemical) labeling index using fresh tissue from 631 node-negative breast cancer patients accessioned at St Luke's Hospital. A modified Nottingham-Bloom-Richardson grade was derived from histopathologic data. Median post-treatment observation was 6.4 years. Agreement on classification of nuclear figures (N=43) was less than good by kappa statistic (kappa=0.38). Grade was moderately reproducible in four trials (N=10,10,19, 10) with CBCTR specimens (kappa=0.50-0.59). Of components of Bloom-Richardson grade, agreement was least for nuclear pleomorphism (kappa=0.37-0.50), best for tubularity (kappa=0.57-0.83), and intermediate for mitotic count (kappa=0.45-0.64). Bloom-Richardson grade was a univariate predictor of prognosis in node-negative St Luke's patients, and was improved when mitotic count was replaced by labeling index (low, mid, or high). When labeling index was added to a multivariate model containing tumor size and vessel invasion, grade was no longer a significant predictor of tumor-specific relapse-free or overall survival. Mitotic index predicted best when intervals were lowered to 0-2, 3-10, and >10 mitotic figures per ten 0.18 mm(2) high-power fields. We conclude that Nottingham-Bloom-Richardson grades remain only modestly reproducible. Prognostically useful components of grade are mitotic index and tubularity. The Nottingham-Bloom-Richardson system can be improved by lowering cutoffs for mitotic index and by counting 20-30 rather than 10 high-power fields. Measurement of proliferation index by immunohistochemically detectable markers will probably give superior prognostic results in comparison to grade.
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              Virtual microscopy in pathology education.

              Technology for acquisition of virtual slides was developed in 1985; however, it was not until the late 1990s that desktop computers had enough processing speed to commercialize virtual microscopy and apply the technology to education. By 2000, the progressive decrease in use of traditional microscopy in medical student education had set the stage for the entry of virtual microscopy into medical schools. Since that time, it has been successfully implemented into many pathology courses in the United States and around the world, with surveys indicating that about 50% of pathology courses already have or expect to implement virtual microscopy. Over the last decade, in addition to an increasing ability to emulate traditional microscopy, virtual microscopy has allowed educators to take advantage of the accessibility, efficiency, and pedagogic versatility of the computer and the Internet. The cost of virtual microscopy in education is now quite reasonable after taking into account replacement cost for microscopes, maintenance of glass slides, and the fact that 1-dimensional microscope space can be converted to multiuse computer laboratories or research. Although the current technology for implementation of virtual microscopy in histopathology education is very good, it could be further improved upon by better low-power screen resolution and depth of field. Nevertheless, virtual microscopy is beginning to play an increasing role in continuing education, house staff education, and evaluation of competency in histopathology. As Z-axis viewing (focusing) becomes more efficient, virtual microscopy will also become integrated into education in cytology, hematology, microbiology, and urinalysis.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, USA )
                1932-6203
                2013
                30 December 2013
                : 8
                : 12
                : e82576
                Affiliations
                [1 ]Symbiant Pathology Expert Centre, Alkmaar, The Netherlands
                [2 ]Symbiant Pathology Expert Centre, Zaandam, The Netherlands
                [3 ]Medical Center Alkmaar, Alkmaar, The Netherlands
                [4 ]Department of pathology, University Medical Centre Utrecht, Utrecht, The Netherlands
                Innsbruck Medical University, Austria
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Conceived and designed the experiments: SAJ MJ. Performed the experiments: SAJ HJS MV. Analyzed the data: SAJ TVP . Contributed reagents/materials/analysis tools: SAJ HJS MV TVP PVD MJ. Wrote the paper: SAJ HSN MJ PVD. Reviewed and edited the final manuscript: HJS MV TVP PVD MJ.

                Article
                PONE-D-13-26290
                10.1371/journal.pone.0082576
                3875418
                24386102
                b429bd1a-0f2c-4b06-b8a2-9deed49cb6bf
                Copyright @ 2013

                This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 20 June 2013
                : 25 October 2013
                Page count
                Pages: 9
                Funding
                These authors have no support or funding to report.
                Categories
                Research Article
                Biology
                Histology
                Molecular Cell Biology
                Chromosome Biology
                Mitosis
                Medicine
                Diagnostic Medicine
                Pathology
                Anatomical Pathology
                Histopathology
                Surgical Pathology
                Clinical Pathology
                General Pathology
                Test Evaluation
                Obstetrics and Gynecology
                Breast Cancer
                Oncology
                Cancers and Neoplasms
                Breast Tumors

                Uncategorized
                Uncategorized

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