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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
Three-dimensional reconstructions producing computer-generated false colour images have played an important role in furthering understanding in many areas in microscopical morphology, both in anatomy and pathology. This review article considers several recent advances made in the understanding of tumour pathology by the use of these computer-generated three-dimensional models. However, three-dimensional reconstructions can be made at a number of levels of resolution - at the level of a whole tissue or organ, at the level of cells or part of a tissue, and at the subcellular level. Examples of three-dimensional reconstructions made at each of these levels of resolution are discussed, with reference to human fetal notochords, bone marrow trephine specimens, and nucleoli from human fibroblasts, Spirogyra grevilleana, mouse Sertoli cells, and pea meristematic cells.
We analyzed the diagnosis, the potentially associated external and clinical features, and the surgical procedures of small pulmonary lesions, especially hamartomas (in relation to peripheral T1 lung carcinomas and lymphoid hyperplasia) in 103 patients who experienced enucleation or resection of pulmonary hamartomas between March 1, 1995 and December 31, 2000. The causes of surgical intervention, presurgical diagnoses, surgical procedures, location, size, and histological compartments were analyzed, as well as clinical features potentially associated with the tumors (alcohol, asbestos, smoking, and chronic lung diseases). Follow up of patients lasted for 5.5 years at maximum. For comparison, 36 patients with peripheral T1 lung carcinomas are included as well as 50 patients with lymphoid hyperplasia. The sex and age distribution of the patients with hamartomas was comparable to that of patients with lymphoid hyperplasia. About 75% of men and 55% of women were heavy smokers, with an average history of 30 and 17 pack years, respectively. In 84% of patients, the lesions were incidentally detected in chest radiographs, whereas 12% of patients underwent thoracic surgery suspicious for intrapulmonary metastases of known extrapulmonary malignancies. Enucleation was performed in 21%, and wedge resection in 77% of patients. At average, hamartomas were smaller than T1 lung carcinomas, but considerably larger in comparison to lymphoid hyperplasia. No recurrent tumors or additionally detected hamartomas were noted during the follow up, and both surgical procedures (enucleation or wedge resection) were identical in curative treatment. All patients with peripherally localized T1 tumors underwent lobectomy. The 3/5 year survival rate was calculated to 69/52%. Lymphoid hyperplasia is of clinical importance for the estimation of prognosis in patients with metastatic disease, as the number of radiologically suggestive metastatic nodules can often be significantly changed due to this entity. Pulmonary hamartomas are benign lesions that display certain clinical associations with malignant lung carcinomas in respect to external risk factors, and to lymphoid hyperplasia. Both surgical procedures (enucleation or wedge resection) can be performed, giving identical results in respect to treatment.
Objective: We report the spectrum of congenital cardiac malformations inautopsied abortions, stillborns and/or neonatal deaths. Prenatal and/or postnatal cytogenetic investigations were carried out in all of the cases. Three fetuses in whom a prenatal diagnosis of 22q11.2 deletion was made by fluorescence in situ hybridization (FISH) are described in detail. Design: During a five year period from 1996 to July, 2000, 132 fetuses with prenatally diagnosed cardiac disease were tested for 22q11.2 microdeletions. Autopsies were performed in 42 cases of abortions, stillborns and/ or neonatal deaths with congenital cardiac defects. Pathological findings, prenatal diagnosis, dysmorphic features and family history are described in detail in three fetuses with CATCH 22. Results: In all cases diagnosed prenatally, cardiac findings were confirmed by autopsy. We found 22q11.2 microdeletion in 5 out of 132 (3.8%) cases with prenatally diagnosed cardiac defects. Because of a severe cardiac defect, and aplasia of the thymus, the pregnancies were terminated in 3 out of these 5 cases diagnosed prenatally with CATCH 22. The cardiac defects in these three fetuses were: common arterial trunk (case 1); tetralogy of Fallot with “absent pulmonary valve syndrome” (case 2), and interruption of the aortic arch between the left common carotid and left subclavian arteries (“Type B”) with ventricular septal defect (case 3). Clinical and cytogenetic examinations of the families showed that, in two cases, the parents were not affected. In the other case, the mother had phenotypic features of the velo-cardio-facial syndrome (VCFS) and a microdeletion 22q11.2. Conclusions: Investigations for microdeletion 22q11.2 by FISH is indicated in addition to conventional karyotyping in fetuses with malformations of the outflow tracts and abnormalities of the arterial trunks. Because of marked phenotypic variability, a test for 22q11.2 microdeletion should be considered in both parents of any child with CATCH 22.
Elucidation of the intricacies of carbohydrate recognition by proteins undoubtedly paves the way for a rational design of carbohydrate-based drugs. Progress towards this goal demands a detailed structural characterisation of the protein target, identification of the ligand determinants essential for recognition and a thorough thermodynamic characterisation of the binding process. To this aim, X-ray crystallography, chemical mapping and calorimetric studies have proved to be highly valuable tools.
Cellular orders in normal tissue and in tumors can be described on the basis of physical terms. Assuming that the peptidase cathepsin B is involved as an essential factor in tumor progression we analyzed the corresponding distribution pattern and determined the entropy of this cell fraction in tumors by syntactic structure analysis using nearest neighbor relationships. Of the 120 surveyed sections from primary lung tumors 62 (51.7 %) were identified with cathepsin B expressing cells. Cathepsin B-positive tumor cells have significantly shorter mean cell-cell distances (p<0.01) and form significantly higher number of tumor cell clusters (p<0.01) when compared with cathepsin B-negative tumor cells. The diameters of the cathepsin B positive tumor cell clusters are increased in moderately and intensively stained tumor cell areas compared to the cathepsin B negative ones (p<0.1 and p<0.05, respectively). The mean cell-cell distance between positive tumor cells was significantly shorter in squamous cell carcinomas (SCC) compared to adenocarcinomas (AC) (p<0.01). We found an increased number of tumor cell clusters in intensively stained areas of SCC compared with AC (p<0.05). Interestingly, in dedifferentiated tumors cells which strongly expressed cathepsin B, have significantly (p<0.01) shorter mean cell-cell distances compared with those well differentiated tumors. In conclusion, cathepsin B positive tumor cells have shorter mean cell-cell distances and form higher number of tumor cell clusters. Our findings indicate that cathepsin B positive tumor cells of primary lung tumors seem to detach as tumor cell clusters instead of single tumor cells. This process appears to be more pronounced in squamous cell carcinomas than in adenocarcinomas.
A fatal case with a primary choriocarcinoma of the lung is presented. The 43 year old women developed pain of the left chest wall for two months. Chest radiographs displayed a small peripherally localized tumor in the left upper lobe measuring 1 cm * 2 cm involving the adjacent pleura. She has been a smoker with about 20 pack years. The histology revealed an undifferentiated carcinoma; the immunohistology tumor cells with strong staining against ß-HCG and (weakly) against alpha-fetoprotein. The diagnosis of a primary choriocarcinoma of the lung was stated. The patient died after twenty-one months due to generalized tumor spread including brain metastases.
Gallbladder carcinoma with osteoblast formation SUMMARY We describe an adenosquamous carcinoma of the gallbladder in a 40 year old male presenting with obstructive jaundice secondary to a bile duct stricture. The bile duct adjacent to the gallbladder was involved solely with adenocarcinoma and this was confirmed with immunohistochemistry using antibodies to ca 19.9. ca 125 and CEA. We postulate that only the adenocarcinoma component of the primary tumour invaded the epithelium of the adjacent bile duct. This led to the progression and involvement of the adjacent bile duct epithelium by the adenocarcinoma component only.
