Diffuse pulmonary hemosiderosis after exposure to pesticides - A case report

This report describes the clinical, radiological, microscopical and ligandohistochemical findings in a 17-year-old woman who suffered from an acute onset of pulmonary hemosiderosis after inhalation of pesticides used for the cultivation of strawberries. She complained of headache, dyspnea, rhinitis, weakness and recurrent severe hemoptysis. Chest radiographs revealed bilateral patchy infiltrates, predominantly in the lower parts of both lungs. The consecutive severe anemia was treated by multiple blood transfusions which were repeated every 4-5 days. Open lung biopsies displayed signs of diffuse hemorrhage with hemosiderin-loaded macrophages, some hyaline membranes, focal fibroid deposits with intermingled histiocytes, mild interstitial fibrosis and focal intra-alveolar calcified bodies surrounded by foreign body giant cells. Analysis of endogenous lectins failed to demonstrate expression of binding capacities for maltose, fucose, mannose; lactose and sialic acid. Neither binding capacities for the macrophage-migration-inhibitory factor nor its presence, as analyzed by labeled sarcolectin, could be detected histochemically. The light microscopical findings are consistent with a longer-lasting diffuse pulmonary hemosiderosis; the presence of calcified bodies and foreign body giant cells (including the ligandohistochemical data) argues for a causal role of inhaled substances. The patient's clinical course improved after cyclophosphamide treatment, which restored her ability to work and released her from the need for recurrent blood transfusions

Review of imaging solutions for integrated quantitative immunohistochemistry in the Pathology daily practice.

Immunohistochemistry (IHC) plays an essential role in Pathology. In order to improve reproducibility and standardization of the results interpretation, IHC quantification methods have been developed. IHC interpretation based in whole slide imaging or virtual microscopy is of special interest. The objective of this work is to review the different computer-based programs for automatic immunohistochemistry and Fluorescence In Situ Hybridization (FISH) evaluation. Scanning solutions and image analysis software in immunohistochemistry were studied, focusing especially on systems based in virtual slides. Integrated scanning and image analysis systems are available (Bacus TMAScore, Dako ACIS III, Genetix Ariol, Aperio Image Analysis, 3DHistech Mirax HistoQuant, Bioimagene Pathiam). Other image analysis software systems (Definiens TissueMap, SlidePath Tissue Image Analysis) can be applied to several virtual slide formats. Fluorescence is the preferred approach in HistoRx AQUA, since it allows for a better compartmentalization of signals. Multispectral imaging using CRi Nuance allows multiple antibodies immunohistochemistry, and different stain unmixing. Most current popular automated image analysis solutions are aimed to brightfield immunohistochemistry, but fluorescence and FISH solutions may become more important in the near future. Automated quantitative tissue microarrays (TMA) analysis is essential to provide high-throughput analysis. Medical informatics standards in images (DICOM) and workflow (IHE) under development will foster the use of image analysis in Pathology Departments

Automated recognition and counting of the immunoreactive neuroendocrine cells in chronic gastritis (the preliminary study).

The paper presents the designed software CAMI (Computerized Analysis of Microscopic Images) for a digital reconstruction of the diversiform glands seen in chronic inflammatory gastric mucosa, and for automated recognition and quantization of the immunoreactive neuroendocrine (NE) cells appearing within mucosal glands. Digital reconstruction of the individual gastric gland is difficult due to variable shapes of the glandular cross-sections. Fifteen gastric biopsy specimens representing chronic gastritis were stained routinely with H+E and immunohistochemically with 3 NE markers: Chromogranin A, Somatostatin and Serotonin. Two expert pathologists counted manually the NE cells with the light microscope in 4 types of glandular cross-sections: round, short- oblique, long- oblique and longitudinal. The automated counting of the NE cells was performed on the digital images presenting the same microscopic areas which were selected for the manual reading. The first step of image analysis was concerned to the cell extraction and recognition of the cytoplasmic immunoreactivity. The unstained nuclei of the NE cells were spotted by the sequential thresholding algorithm combined with the artificial neural network of Support\Vector Machine (SVM) type. The second step of image analysis comprised reconstruction of the glands. The presumed shape of each gastric gland was defined by the cellular lining of viewed glandular cross-section. The designed algorithm for gland reconstruction was based on the cell masks. The third step of analysis dealt the cell counting. Every recognized gland with the face cells was used for the NE cell evaluation. The results of the automated quantization compared with manual counting results for the number of NE cells showed high concordance in 3 types of glandular cross-sections: round, short- and long- oblique. A difference noticed in the results of the longitudinal glands should be verified in the extended study. The designed software CAMI is more adequate for the gland recognition with an discontinuous gland face seen in the immunohistochemical digital images, which appear to be a difficult problem for the accurate automated analysis of the cellular component of glands

Predicting Breast Cancer Response to Neoadjuvant Chemotherapy Using Pretreatment Diffuse Optical Spectroscopic-Texture Analysis

Purpose: Diffuse optical spectroscopy (DOS) has been demonstrated capable of monitoring response to neoadjuvant chemotherapy (NAC) in locally advanced breast cancer (LABC) patients. In this study, we evaluate texture features of pre-treatment DOS functional maps for predicting LABC response to NAC. Methods: LABC patients (n = 37) underwent DOS-breast imaging before starting neoadjuvant chemotherapy. Breast-tissue parametric maps were constructed and texture analyses were performed based on grey level co-occurrence matrices (GLCM) for feature extraction. Ground-truth labels as responders (R) or non-responders (NR) were assigned to patients based on Miller-Payne pathological response criteria. The capability of DOS-textural features computed on volumetric tumour data before the start of treatment (i.e. “pre-treatment”) to predict patient responses to NAC was evaluated using a leave-one-out validation scheme at subject level. Data were analysed using a logistic regression, naïve Bayes, and k-nearest neighbour (k-NN) classifiers. Results: Data indicated that textural characteristics of pre-treatment DOS parametric maps can differentiate between treatment response outcomes. The HbO2-homogeneity resulted in the highest accuracy amongst univariate parameters in predicting response to chemotherapy: sensitivity (%Sn) and specificity (%Sp) were 86.5 and 89.0%, respectively and accuracy was 87.8%. The highest predictors using multivariate (binary) combination features were the Hb-Contrast + HbO2-Homogeneity which resulted in a %Sn/%Sp = 78.0/81.0% and an accuracy of 79.5%. Conclusions: This study demonstrated that pre-treatment tumour DOS-texture features can predict breast cancer response to NAC and potentially guide treatments

Comparison of the Manual, Semiautomatic, and Automatic Selection and Leveling of Hot Spots in Whole Slide Images for Ki-67 Quantification in Meningiomas

Background. This paper presents the study concerning hot-spot selection in the assessment of whole slide images of tissue sections collected from meningioma patients. The samples were immunohistochemically stained to determine the Ki-67/MIB-1 proliferation index used for prognosis and treatment planning. Objective. The observer performance was examined by comparing results of the proposed method of automatic hot-spot selection in whole slide images, results of traditional scoring under a microscope, and results of a pathologist’s manual hot-spot selection. Methods. The results of scoring the Ki-67 index using optical scoring under a microscope, software for Ki-67 index quantification based on hot spots selected by two pathologists (resp., once and three times), and the same software but on hot spots selected by proposed automatic methods were compared using Kendall’s tau-b statistics. Results. Results show intra- and interobserver agreement. The agreement between Ki-67 scoring with manual and automatic hot-spot selection is high, while agreement between Ki-67 index scoring results in whole slide images and traditional microscopic examination is lower. Conclusions. The agreement observed for the three scoring methods shows that automation of area selection is an effective tool in supporting physicians and in increasing the reliability of Ki-67 scoring in meningioma

Computer-assisted quantification of caix membrane immunoreaction destined for the clear cells in renal carcinoma. A pilot study.

Introduction/ Background Carbonic Anhydrase IX [CAIX] has been considered as a candidate prognostic factor in clear-cell renal carcinoma [CCRC], however the supporting evidence is conflicting. CAIX is strongly induced by hypoxia via HIV-1α, and in CCRC via mutations to the VHL gene. CAIX expression could be identify as an immunohistochemical predictor of CCRC patients outcome but the published studies related to the patients prognosis have based on the diverse quantification protocols of CAIX expression (TMAs vs. whole tissue section; semiquantitative vs. computerised image analysis; with/without intensity scoring; with various software). The available commercial image analysis tools are mainly for general purpose e.g. software for breast carcinoma HER2 membrane immunoreaction has been used in various tumour tissue studies. However the cytological images of CCRC and breast carcinoma show essential differences related to the nuclei (size, outlines, intracellular location) and nuclear/cytoplasmic proportion which could influence the measurement credibility in maladjusted algorithm. Aims The aim of our study was to evaluate an algorithm for quantification of the membranous CAIX expression specifically dedicated to CCRC (“snake variant”) in comparative analysis to applied HER2 breast cancer algorithm for CCRC. Methods In the quantitative analysis of the specimen, the image processing follows: recognition of the cell nuclei; segmentation of the immunoreactive cell membranes; the assignment of the membrane segments to an individual cell. The last step is challenging for analysis due to frequent discontinuities in membranous immunoreaction, great variability of cellular counters and intracellular nuclei location. Because the classical watershed method for the individual cell separation is insufficient, the snake active contour method was applied, starting from each nucleus outline. The built gradient image allowed to select the most adequate parameters in the snake adaptation process. The recognized snake represents the membrane associated with the particular cell. The material includes records of 39 patients with the histopathologically verified diagnosis of CCRC who had nephrectomy (between 2009-2011) and were treated with tyrosine kinases agents (the Clinic of Oncology registry). 74% (29 out 39) of patients presented stage I - T1 N0; 20,5% - stage III and 5,4% stage TII. The formalin-fixed tissue sections of the resected CCRCs (the Pathology Department registry) were immunostained for CAIX protein using CAIX antibody (clone NB100-417) (Antibodies-online GmbH) with EnVisionTM (DAKO) according to the manufacture recommendations. The representative digital images were selected from each Whole Slide Image (scanned with Aperio, under 20x) and were assessed automatically by 3 independent observers using two algorithms: “snake variant” and “breast HER2”. The extend of staining (percentage) was scored in the 10% intervals of CAIX positive carcinomatous cells and the intensity of immunoreaction was evaluated in 3 grade scale (1-3). ResultsThe obtained results have been under investigation for the intra- and inter-observer accuracy as well as for the comparative data analysis of both types of algorithm. The statistical analysis has been incorporated. This approach explores a new possibility of the computerised quantitative estimation of the membrane CAIX immunoreaction destine

Evaluation of HER2 Gene Amplification in Breast Cancer Using Nuclei Microarray in Situ Hybridization

Fluorescence in situ hybridization (FISH) assay is considered the “gold standard” in evaluating HER2/neu (HER2) gene status. However, FISH detection is costly and time consuming. Thus, we established nuclei microarray with extracted intact nuclei from paraffin embedded breast cancer tissues for FISH detection. The nuclei microarray FISH (NMFISH) technology serves as a useful platform for analyzing HER2 gene/chromosome 17 centromere ratio. We examined HER2 gene status in 152 cases of invasive ductal carcinomas of the breast that were resected surgically with FISH and NMFISH. HER2 gene amplification status was classified according to the guidelines of the American Society of Clinical Oncology and College of American Pathologists (ASCO/CAP). Comparison of the cut-off values for HER2/chromosome 17 centromere copy number ratio obtained by NMFISH and FISH showed that there was almost perfect agreement between the two methods (κ coefficient 0.920). The results of the two methods were almost consistent for the evaluation of HER2 gene counts. The present study proved that NMFISH is comparable with FISH for evaluating HER2 gene status. The use of nuclei microarray technology is highly efficient, time and reagent conserving and inexpensive

Identifying determinants of adherence to adjuvant endocrine therapy following breast cancer: A systematic review of reviews

BACKGROUND: In oestrogen-receptor positive breast cancer, daily oral adjuvant endocrine therapy (ET) for at least 5 years significantly reduces risks of recurrence and breast cancer-specific mortality. However, many women are poorly adherent to ET. Development of effective adherence support requires comprehensive understanding of influences on adherence. We undertook an umbrella review to identify determinants of ET adherence. METHODS: We searched PubMed, Embase, CINAHL, PsycINFO, Cochrane and PROSPERO (inception to 08/2022) to identify systematic reviews on factors influencing ET adherence. Abstracted determinants were mapped to the World Health Organization's dimensions of adherence. Reviews were quality appraised and overlap assessed. RESULTS: Of 5732 citations screened, 17 reviews were eligible (9 quantitative primary studies; 4 qualitative primary studies; 4 qualitative or quantitative studies) including 215 primary papers. All five WHO dimensions influenced ET non-adherence: The most consistently identified non-adherence determinants were patient-related factors (e.g. lower perceived ET necessity, more treatment concerns, perceptions of ET 'cons' vs. 'pros'). Healthcare system/healthcare professional-related factors (e.g. perceived lower quality health professional interaction/relationship) were also important and, to a somewhat lesser extent, socio-economic factors (e.g. lower levels of social/economic/material support). Evidence was more mixed for medication-related and condition-related factors, but several may be relevant (e.g. experiencing side-effects, cost). Potentially modifiable factors are more influential than non-modifiable/fixed factors (e.g. patient characteristics). CONCLUSIONS: The evidence-base on ET adherence determinants is extensive. Future empirical studies should focus on less well-researched areas and settings. The determinants themselves are numerous and complex in indicating that adherence support should be multifaceted, addressing multiple determinants