New automated image analysis method for the assessment of Ki-67 labeling index in meningiomas.

Many studies have emphasised the importance of Ki-67 labeling index (LI) as the proliferation marker in meningiomas. Several authors confirmed, that Ki-67 LI has prognostic significance and correlates with likelihood of tumour recurrences. These observations were widely accepted by pathologists, but up till now no standard method for Ki-67 LI assessment was developed and introduced for the diagnostic pathology. In this paper we present a new computerised system for automated Ki-67 LI estimation in meningiomas as an aid for histological grading of meningiomas and potential standard method of Ki-67 LI assessment. We also discuss the concordance of Ki-67 LI results obtained by presented computerized system and expert pathologist, as well as possible pitfalls and mistakes in automated counting of immunopositive or negative cells. For the quantitative evaluation of digital images of meningiomas the designed software uses an algorithm based on mathematical description of cell morphology. This solution acts together with the Support Vector Machine (SVM) used in the classification mode for the recognition of immunoreactivity of cells. The applied sequential thresholding simulated well the human process of cell recognition. The same digital images of randomly selected tumour areas were parallelly analysed by computer and blindly by two expert pathologists. Ki-67 labeling indices were estimated and the results compared. The mean relative discrepancy between the levels of Ki-67 LI by our system and by the human expert did not exceed 14% in all investigated cases. These preliminary results suggest that the designed software could be an useful tool supporting the diagnostic digital pathology. However, more extended studies are needed for approval of this suggestion

Study on breast carcinoma Her2/neu and hormonal receptors status assessed by automated images analysis systems: ACIS III (Dako) and ScanScope (Aperio).

Her-2/neu is overexpressed in 20-30% of breast cancer patients and is associated with a more aggressive disease. Identification of Her-2/c-erbB-2-neu overexpression is based on immunohistochemical [ihc] detection of protein and/or gene amplification in fluorescence in situ hybridization test (FISH). Also Estrogen receptors [ER] and Progesterone receptors [PR] are the prognostic and predictive biomarkers, recently analysed by ihc methods. Subjective, manual scoring of the ihc Her-2/neu expression and expression of the ER/PR reported as the percentage of immunopositive cells are the most common mode of interpretation among pathologists. Automated microscopy and computerised processing have provided increased accuracy in quantification and standardisation. The aims of our study were: to evaluate the scoring reproducibility of Her-2 /neu ihc expression tested by two automated systems: ACIS (Dako) and ScanScope (Aperio); to estimate the ER/PR expression in ihc staining methods with different anti-ER/anti-PR antibodies (the monoclonal and the ER/PR pharmDx TM Kit) by the ACIS system. Her-2/neu ihc expression was measured in 114 primary invasive breast carcinomas by the manual and the automated scoring (ACIS and Aperio system). 106 slides stained ihc with two types of anti-ER/anti-PR antibodies entered the quantisation. The results of our investigations showed very high reproducibility of Her-2/neu scores in intra- and interobserver analysis by ACIS evaluation. The major concordance was present in strong 3+ ihc cases; very small discordance was shown by cases with low expression of Her-2/neu. The accuracy of scoring by the Aperio was little lower in comparison to ACIS but it might result from the smaller and variable series of samples analysed by Aperio. The concordance in scoring of two automated systems was 86.5% (

Morphometric and functional abnormalities of kidneys in the progeny of mice fed chocolate during pregnancy and lactation.

Even most commonly consumed beverages like tea, coffee, chocolate and cocoa contain methylxanthines, biogenic amines and polyphenols, among them catechins, that exhibit significant biological activity and might profoundly affect the organism homeostasis. We have previously shown that 400 mg of bitter chocolate or 6 mg of theobromine added to the daily diet of pregnant and afterwards lactating mice affected embryonic angiogenesis and caused bone mineralization disturbances as well as limb shortening in 4-weeks old offspring. The aim of the present study was the morphometric and functional evaluation of kidneys in the 4-weeks old progeny mice fed according to the protocol mentioned above. Progeny from the mice fed chocolate presented considerable morphometric abnormalities in the kidney structure, with the lower number of glomeruli per mm2 and their increased diameter. Moreover, higher serum creatinine concentration was observed in that group of offspring. No morphometric or functional irregularities were found in the progeny of mice fed theobromine. Abnormalities demonstrated in the offspring of mice fed chocolate are not related to its theobromine content. Consequently, identification of active compound(s) responsible for the observed effects is of vital importance

Accuracy of a remote quantitative image analysis in the whole slide images

The rationale for choosing a remote quantitative method supporting a diagnostic decision requires some empirical studies and knowledge on scenarios including valid telepathology standards. The tumours of the central nervous system [CNS] are graded on the base of the morphological features and the Ki-67 labelling Index [Ki-67 LI]. Various methods have been applied for Ki-67 LI estimation. Recently we have introduced the Computerized Analysis of Medical Images [CAMI] software for an automated Ki-67 LI counting in the digital images

Digital pathology in personalized cancer therapy

The development of small molecule inhibitors of growth factor receptors, and the discovery of somatic mutations of the thyrosine kinase domain, have resulted in new paradigms for cancer therapy. Digital microscopy is an important tool for surgical pathologists. The achievements in the digital pathology field have modified the workflow of pathomorphology labs, enhanced the pathologist’s role in diagnostics, and increased their contribution to personalized targeted medicine. Digital image analysis is now available in a variety of platforms to improve quantification performance of diagnostic pathology. We here describe the state of digital microscopy as it applies to the field of quantitative immunohistochemistry of biomarkers related to the clinical personalized targeted therapy of breast cancer, non-small lung cancer and colorectal cancer: HER-2, EGFR, KRAS and BRAF genes. The information is derived from the experience of the authors and a review of the literature. (<i>Folia Histochemica et Cytobiologica 2011; Vol. 49, No. 4, pp. 570–578</i>

Use of the virtual slide and the dynamic real-time telepathology systems for a consultation and the frozen section intra-operative diagnosis in thoracic/pulmonary pathology.

We report the results of a study designed for assessment of the diagnostic accuracy and usability of internet-based digital microscopy: the dynamic real-time telepathology system (Coolscope) and the Virtual microscopy (Aperio Scan Scope) system, in the context of pulmonary pathology. The systems were implemented to the routine pulmonary pathology workflows and used for the intra-operative frozen-section primary diagnosis as well as for the secondary (consultative) diagnosis. The histological material presented for the teleconsultations included the samples of lung parenchyma, bronchial biopsy and resected lung/bronchi tumours. For the primary diagnosis 4 categories of material can be distinguished (304 samples): 1) the frozen sections of lung tumours, resected bronchial margins and lymph nodes; 2) fine needle aspiration [FNA] biopsies (TBNA; EBUS-TBNA, EUS-FNA; 3) oligobiopsies of bronchus, oesophagus, skin; and 4) exfoliative cytology. The telepathology diagnoses compared with conventional light microscopy diagnoses showed very high concordance for the Coolscope and Aperio Virtual Slide modality: 87.5% and 100%, respectively - within the group of teleconsultations. For the frozen sections, the primary telediagnoses were concordant with the light microscopy paraffin sections diagnoses in 100% for Aperio; and in 97.5% for Coolscope. An excellent agreement (100%) was seen in the telediagnoses and conventional slides diagnoses for FNA, oligobiopsies and cytology - for both telepathology systems. These results provide some encouragement for the implementation of Coolscope and virtual slide-based telepathology (Aperio) system to the routine histopathological diagnostics