Electronic Journal of Pathology and Histology - Volume 8, issue 1-4

Authors: Collan, Yrjo | Kronqvist, P. | Kuopio, T.

Article Type: Research Article

Abstract: Background Quantitative pathology deals with graded or continuous features. Often these features are prognosticators, and able to tell something about the outcome of disease. Many features are also expected to be predictors of treatment response, especially in association with drug therapy. The basicly intuitive knowledge that the expression of estrogen and progesterone receptors on the surface of cancer cells could reflect good response during tamoxifen or toremifen treatment is much valued in clinical practice, as is the overexpression of erbB2 receptor antigen on the cell surface in potentially predicting a good response during trastuzumab treatment. Material and methods Prognostication, …and prediction of therapy outcome is not always perfect, even with multivariate methodology. Because of this, we have looked for a method which could extract most of the prognostic value from basicly quantitative histological or immunohistochemical features. For this purpose we have applied various morphometric tests, and studied various antibodies, including antibodies against cystatin A, bcl-2, erb-B2, and E-cadherin in immunohistochemistry. In immunohistochemistry the staining can be subjectively graded, but usually the immunohistochemical staining index turns out to be the best prognosticator. In our studies we tried to define the prognostically most valuable cutpoints, both in subjective grading and in calculating immunohistochemical staining indices. Cutpoint optimization was done with the help of the khi-square test. After optimization, the prognostic evaluation, in the form of a survival study was carried out. Corresponding studies were carried out on various morphometric features, associated with the development of a morphometric grading system for breast cancer. Results Many quantitative features had a cutpoint or cutpoints which showed highly significant survival difference between patients on the two sides of the cutpoint Some cutpoints were especially significant and associated with prominent peaks in the khi-square curve or ditches in the p-value curve. These cutpoints were chosen as optimal cutpoints for further evaluation. In the following survival analysis the cutpoint presented the best alternative for the survival analysis. Prognostic comparison of the features also appeared more reliable when the optimal cutpoint was known. However, also after these studies the final prognostic conclusions should be based on an independent material, suitable for corresponding analysis. Conclusions A reliable analysis of potential prognosticators or predictors can be based on optimization of the decision cutpoint. After the procedure different prognosticators can be compared reliably. In clinical implementation separate the thinking of group statistics should be changed to thinking of diagnostic statistics. Show more

Keywords: breast cancer, prognosis, diagnosis, grading, khi square, univariate methods, multivariate methods, prognosticators, predictors, survival analysis, histopathology, clinical chemistry

Citation: Electronic Journal of Pathology and Histology, vol. 8, no. 1-4, pp. 01-01, 2003

Authors: Ioachim, Elli | Charchanti, Antonia | Charalabopoulos, Kostas | Tsanou, Helena | Briasoulis, Evangelos | Karavasilis, Vassilis | Pavlidis, Nikolaos | Agnantis, Niki J.

Article Type: Research Article

Abstract: Microvessel density (MVD) was analyzed for associations with clinical and pathological factors as well as with other potential known prognostic factors such as: steroid receptor content (ER, PgR), p53 and proliferation associated indices (Ki-67, PCNA). In the present study microvascular quantification was undertaken on 145 cases of breast carcinoma after immunohistochemical staining of tumor vessel, using polyclonal antibody to factor VIII related antigen. Microvessel quantification was performed at x400 magnification in the three most vascular areas of the tumors (hot spots) usually located at the periphery or growing front of the tumor. In addition, a semi-quantitative evaluation of the number …of vessels per mm2 of highest intratumoral microvessel density (MVD) was performed by use of an image analysis system. Significant correlation was observed in MVD by computerized analysis and by light microscopy counting (p=0.02). Survival analysis showed increased mortality risk associated with low MVD estimated by both methods (p-0.043 and p=0.041 respectively) including node-negative and node positive subsets. ). In addition low MVD was correlated with recurrence (p=0.02) and metastatic disease (p=0.0016) in the cases estimated by light microscopy. However in multivariant analysis, MVD was independently correlated with relapse-free and over patients survival. MVD was higher in lobular than in ductal cell carcinoma (p=0.015). MVD was positive correlated with estrogen receptor status (p=0.04) and inversely with tumor size (p=0.004). No association was found with tumor grade and lymph node status The results of the present study show that the MVD (determined at a microscopic level or by image analysis) correlated with better prognostic parameter maybe due to better responded to treatment. Furthermore, MVD in addition to breast cancer heterogeneity, could be reflects different phases of tumor growth. Show more

Keywords: Angiogenesis, ER, PgR, p53, Ki-67, PCNA, breast cancer

Citation: Electronic Journal of Pathology and Histology, vol. 8, no. 1-4, pp. 02-02, 2003

Authors: Malamou-Mitsi, Vassilis | Zachariou, Christianna | Tzallas, Sotiris | Tavoulari, Sotiria | Paraskevaidis, Evangelos | Agnantis, Niki J.

Article Type: Research Article

Abstract: The identification and typing of HPVs (Human Papilloma Viruses) has become increasingly challenging, due to the numerous viral types that must be detected and their role in the process of cervical neoplasia. In the present study we used a combination of PCR amplification method and RFLP analysis in order to identify established or presumed novel HPV-types in various cervical lesions and consequently to evaluate its effectiveness in archival histopathological tissue samples. Forty two cases with histologic diagnosis of "pure" HPV-lesions [n=3], low grade intraepithelial lesions-LGSIL [n=16] and high grade intraepithelial lesions-HGSIL [n=23] were studied. Overall HPV-DNA was detected in 30 …out of 42 cases analyzed [Detection rate: 71.4%]. High- risk HPV-types were detected in 15 of 23 HGSILs [65.1%], in 11 of 16 LGSILs [68.7%] and in one of 3 cases of "pure" HPV-lesions [33.3%]. Low risk HPV-types were detected in 1 of 23 HGSILs [4,3%] and in 1 of 16 LGSILs [6.2%]. One possibly novel HPV-type was detected in one case of LGSIL. HPV-typing by RLFP analysis was successful in 13 out of 30 positive cases [43.3%]. HPV-16 was identified in 6 cases [5 of HGSIL and 1 of LGSIL], HPV-31 in 3 cases [1 of HGSIL with microinvasion and 2 of LGSIL], HPV-33 in 1 case of HGSIL, HPV-56 in 1 case of LGSIL, HPV-61 in 1 case of HPVlesion and one possibly novel HPV-type in a case of LGSIL. This method could facilitate the sensitive identification of a broad spectrum of known and novel genital HPVs and may serve as an adjunct to existing traditional hybridization-based methods. Show more

Keywords: HPV typing, PCR, RFLP analysis, cervical intraepithelial lesions

Citation: Electronic Journal of Pathology and Histology, vol. 8, no. 1-4, pp. 03-03, 2003

Authors: Charhanti, Antonia | Ioachim, Elli | Eyden, Brian | Agnantis, Niki J.

Article Type: Research Article

Abstract: Amorphous matrices related with basement membrane have been described before, in peri-vascular locations in malignancies, reactive lesions and normal cerebral cortex and on the surfaces of benign tumoral fibroblasts. They appear to be compartments comprising the proteins normally found in an organised (laminate) lamina (lamina densa/lamina lucida), but not in fact organised in that manner, and have a disorganized a amorphous appearance. By light microscopy immunohistochemistry, BM of vessels and epithelial cells stained positively for laminin and collagen IV, two of the main proteins characterizing a conventional basal lamina. This paper discusses the possibility that ultrastructural findings in …correlation with those of immunohistochemistry supply more information for the biological behavior of breast cancer that seems to be influenced by the pattern of organization of BM proteins. These models of organization may be involved in the metastatic process. Show more

Keywords: amorphous matrix in breast cancer

Citation: Electronic Journal of Pathology and Histology, vol. 8, no. 1-4, pp. 04-04, 2003

Authors: Mazzucchelli, Roberta | Santinelli, Alfredo

Article Type: Research Article

Abstract: A Bayesian belief network (BBN), a diagnostic decision support system, enables the processing of our knowledge of histopathology expressed in descriptive linguistic terms, words and concepts. The aim of this study was to evaluate the contribution of a BBN to the improvement of observer agreement and certainty level in grading urothelial papillary neoplasms. The study was based on 40 cases of non-invasive urothelial papillary tumours subdivided according to the WHO 1973 classification. There were ten urothelial papillomas (UP), ten grade 1 papillary carcinomas (G1), ten grade 2 papillary carcinomas (G2), and ten grade 3 papillary carcinomas (G3). Five consecutive …sessions were held with three observers at a time interval of at least two weeks. Three of these sessions (A, B and D) were based on the morphological evaluation of the specimens with a conventional light microscope only and in remaining twosessions (C and E) a BBN was used in addition to the microscope. Observer agreement was seen in 60% (RMa), 55% (PC) and 65% (MSt) of cases, respectively, in session A where a synthetic approach to decisionmaking was adopted. The level of subjective certainty was “certain” in 20% of cases (a two tier system was adopted: certain vs less certain). A better observer agreement - 70% (RMa), 68% (PC) and 72% (MSt) of cases - was present in session B where an analytical approach based on the evaluation of a series of morphological features was used. The level of certainty was “certain” in 50% of cases. A further increase in the observer agreement - 85% (RMa), 82% (PC) and 86% (MSt) of cases – with high levels of certainty or belief was seen in session C where a BBN was utilised. A drop in the level of agreement - 60% (RMa), 62% (PC) and 65% (MSt) of cases - and a decrease in certainty were seen in session D where the observers were left free to evaluate morphologically the cases without the constrain of either a synthetic or analytical approach. In session E, agreement increased to 83% (RMa), 81% (PC) and 84% (MSt), respectively, when the BBN was used again. Conventional morphological evaluation of urothelial papillary neoplasms is affected by observer variability and, in many instances, by diagnostic uncertainty. Improvement in observer agreement and certainty level can be achieved with a BBN used in conjunction with a light microscope. Show more

Keywords: Bayesian network, urothel papilloma, morphology, analytical approach

Citation: Electronic Journal of Pathology and Histology, vol. 8, no. 1-4, pp. 05-05, 2003

Authors: Montironi, Rodolfo | Mazzucchelli, Roberta

Article Type: Research Article

Abstract: The aim of the study was to investigate immunohistochemically the expression of vascular endothelial growth factor (VEGF) and its correlation with the pattern of capillary architecture in prostate cancer and high-grade prostatic intraepithelial neoplasia (PIN), in untreated and androgen-ablated patients. Forty-five patients who underwent radical prostatectomy (RP) for localized prostate carcinoma were recruited for this study. The study population included 2 groups: 35 patients who did not receive chemo-, hormone or radiation therapy before surgery, and 10 patients who were under complete androgen blockade (CAB) for three months at the time of operation. VEGF was examined by immunohistochemistry and its …tissue expression was compared with the pattern of capillary architecture evaluated by immunostaining the endothelial antigen CD34. The relationship of VEGF expression to chromogranin A positive (e.g., neuroendocrine) cells was investigated. Significant levels of VEGF are present in prostate cancer and in a population of PIN lesions, the expression being highest in association with NE cells and correlated with an altered pattern of vascularization. The VEGF expression is downregulated by hormonal manipulation, except in the population of NE cells. All this indicates that VEGF may contribute to the establishment, progression and regression of prostate neoplasia. Show more

Keywords: Prostate neoplasia, vascular endothelial growth factor, prostate intraepithelial neoplasia, complete androgen blockade

Citation: Electronic Journal of Pathology and Histology, vol. 8, no. 1-4, pp. 06-06, 2003

Authors: Szymas, Janusz

Article Type: Research Article

Abstract: Telepathology is the diagnostic work of a pathologist at a distance. It often requires specific technical solutions. These include telemicroscopes, telepathological systems, proper telecommunicaton links and computer graphic terminals. Development of computer science, digitalization of macro- and microscopic pictures, dissemination of microcomputers permit the use of electronic data interchange for diagnosis or education in pathology. Standard technologies like HTML, JavaScript and digitalized static pictures may be used for the information service. The information service on Web server can be used for the course of pathology during the semester curriculum. The two main models of telepathology (static versus dynamic), their applications, …the use of telepathology at the Department of Pathology of the University of Medical Sciences in Poznan for didactic purposes and some future aspects will be discussed. TELEMIC telepathology system is used in a local area network, which connects classrooms with the laboratory. It is therefore possible to include this technique into education and diagnostic training. A virtual slide is a fully digital representation of the histological specimen including complete series of static images taken at all magnifications. Implementation of this in an education system requires two modules. The first implementation allows the acquisition of images of the entire specimen. The second module permits the examination of virtual slide by intuitive and user-friendly interface. In combination with clinical and radiological data the construction of the virtual case is possible. By viewing a virtual slide it is possible to search for areas of interest and to examine the slide at various magnification. Such activity needs international cooperation, dedicated international servers, reference databases and telepathology networks, which allow continuous education in pathology. It allows the creation of multimedial database, archiving of data and exchange of information. Show more

Keywords: Telepathology, teleeducation, quality assurance, static telepathology, dynamic telepathology

Citation: Electronic Journal of Pathology and Histology, vol. 8, no. 1-4, pp. 07-07, 2003

Authors: Kayser, Klaus

Article Type: Research Article

Abstract: Static DNA cytometry is one of the few quantitative techniques, which have been successfully applied for diagnostic pathology. Having introduced and defined minimum requirements for this technique, it plays now-a-days a significant role in confirming or ruling out malignant tumours of various origins. In this article, some basic aspects of this technique are described. The following prerequisites are used: a) all nuclei of “normal” human cells which display the same position within the cell cycle contain the same amount of DNA; b) all human cells repeat the cell cycle with the same velocity; c) the cell cycle can roughly be …divided into threeseparated stages, namely resting cells (G0), multiplication stage (S-phase), and reorganization stage (mitosis, doubled DNA content); d) cells undergoing degradation or apoptosis are negligible for reference cells. By use of these prerequisites, the following parameters can be derived: a) the number of reference cells at a cell cycle stage I (1,2,3) is proportional to the duration of the cell cycle stage I; b) the cell cycle stage I of the reference cells can be calculated from the absolute DNA content and the frequency distribution of the cell cycle stages (I-III); the absolute DNA content of reference cells can be computed from the frequency distribution of reference cells within the different cell cycles. For non-reference cells, similar derivatives hold true: a) the amount of additive/missing DNA in non-reference cells is proportional to the amount of DNA of reference cells measured at the same cell cycle stage; b) the additive/missing amount of DNA of non-reference cells can be derived from the DNA distribution of reference cells and that of non-reference cells. The significance of these derivatives for application of DNA cytometry is discussed. Show more

Keywords: Static DNA cytometry, cell cycle, standardization, DNA content

Citation: Electronic Journal of Pathology and Histology, vol. 8, no. 1-4, pp. 08-08, 2003

Authors: Slodkowska, Janina | Radomski, Piotr | Remiszewski, Pawel | Zych, Jacek | Androsiuk, Wojciech | Rowinska-Zakrzewska, Ewa

Article Type: Research Article

Abstract: BACKGROUND: The studies on the therapeutic effect on the disease pattern and morphologic heterogeneity of small cell lung carcinoma (SCLC) provide some explanation for unresponsive behaviour of SCLC to chemotherapy. The aim of the study was to assess the distribution of SCLC disease and variation of the histological SCLC subtypes in relation to the therapy, in post-mortem specimens. METHODS: The autopsies of 85 SCLC patients (pts) treated with various therapy regimes, 15 untreated SCLC pts and 85 diagnostic biopsy samples of treated SCLC pts were reviewed to assess the distribution of the disease and the histological subtypes of SCLC …(IASLC classification). RESULTS: The modalities of therapy were related to status of the primary tumour. The chemo-radiotherapy significantly decreased the presence of macroscopic primary focus in comparison with chemotherapy alone (p=0.03); and significantly increased the frequency of the residual tumour (p=0.026). Six of 45 (13%) pretreated pure SCLC changed their morphology into mixed SCLC or combined SCLC in autopsy postreated tumours. The heterogeneity in distribution of pure SCLC, mixed SCLC and combined SCLC subtypes was present in comparative analysis of primary tumour and its metastases. CONCLUSION: The modalities of therapy influence status of the locoregional disease but do not influence the metastatic pattern of SCLC. The findings indicate that pretreated pure SCLC can progress to mixed SCLC or combined SCLC in postreated cancer, exhibiting a mixed differentiation in the metastases. Show more

Keywords: small cell lung cancer, therapy, histological subtypes, autopsy

Citation: Electronic Journal of Pathology and Histology, vol. 8, no. 1-4, pp. 09-09, 2003

Authors: Gollowitsch, Franz | Berger, Anton | Popper, Helmut H.

Article Type: Research Article

Abstract: A basaloid cell carcinoma of the stomach was detected in an 80-years old male patient. The carcinoma was found to form amyloid P and induced an amyloid P deposition in the inguinal area. Within the carcinoma cytokeratin filaments were gradually replaced by amyloid P, which subsequently resulted in ghost cell formation, simulating pilomatricoma-like appearance. Extracellular amyloid deposition elicited a desmoplastic stroma reaction and metaplastic bone formation. To the best of our knowledge this is the first case of a basaloid cell carcinoma of the stomach with ghost cells, and amyloid deposition.

Keywords: Amyloidosis, Gastric basaloid cell carcinoma

Citation: Electronic Journal of Pathology and Histology, vol. 8, no. 1-4, pp. 10-10, 2003

Authors: Kayser, Klaus

Article Type: Book Review

Citation: Electronic Journal of Pathology and Histology, vol. 8, no. 1-4, pp. 11-11, 2003