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Biomedical subjects

Panagiota Ravazoula

Publications and source records attributed to Panagiota Ravazoula.

16 recordsLinked to original sources

Cell-nuclear data reduction and prognostic model selection in bladder tumor recurrence.

OBJECTIVE: The paper aims at improving the prediction of superficial bladder recurrence. To this end, feedforward neural networks (FNNs) and a feature selection method based on unsupervised clustering, were employed. MATERIAL AND METHODS: A retrospective prognostic study of 127 patients diagnosed with superficial urinary bladder cancer was performed. Images from biopsies were digitized and cell nuclei features were extracted. To design FNN classifiers, different training methods and architectures were investigated. The unsupervised k-windows (UKW) and the fuzzy c-means clustering algorithms were applied on the feature set to identify the most informative feature subsets. RESULTS: UKW managed to reduce the dimensionality of the feature space significantly, and yielded prediction rates 87.95% and 91.41%, for non-recurrent and recurrent cases, respectively. The prediction rates achieved with the reduced feature set were marginally lower compared to the ones attained with the complete feature set. The training algorithm that exhibited the best performance in all cases was the adaptive on-line backpropagation algorithm. CONCLUSIONS: FNNs can contribute to the accurate prognosis of bladder cancer recurrence. The proposed feature selection method can remove redundant information without a significant loss in predictive accuracy, and thereby render the prognostic model less complex, more robust, and hence suitable for clinical use.

Algorithms↗

Non-palpable breast carcinomas: correlation of mammographically detected malignant-appearing microcalcifications and epidermal growth factor receptor (EGFR) family expression.

Malignant-appearing microcalcifications (MAMCs) represent one of the earliest mammographic findings of non-palpable breast carcinomas (NPBCs). In the present study, we have evaluated the expression of all EGFR family members in NPBCs and its possible association with MAMCs. Three hundred and fifty patients with non-palpable suspicious breast lesions detected during screening mammography were studied. EGFR family proteins' expression was found to be present since the preclinical phase of breast carcinomas and was strongly correlated (except HER-3) with MAMCs. The co-expression pattern of EGFR family members combined with other molecular prognostic factors and the mammographic appearance might predict the natural history of NPBCs.

Biomarkers, Tumor↗

Distal embolism during percutaneous revascularization of infra-aortic arterial occlusive disease: an underestimated phenomenon.

PURPOSE: To investigate distal embolism during endovascular procedures of the infra-aortic arteries by utilizing a commercial filter basket and unveil any correlation between the baseline clinical and procedural variables and the histopathological findings of the collected particles. METHODS: In a prospective study, 48 patients (37 men; mean age 70.8+/-7.8 years, range 50- 83) underwent endoluminal therapy of infra-aortic lesions (stenosis >75% or occlusion; mean lesion length 52.2+/-38.0 mm) with standard endovascular procedures. A nitinol filter basket (n=50) was employed for distal protection. The collected particles were histopathologically analyzed. The harvested specimens were quantified after digital image post processing. RESULTS: Procedural success of filter-protected revascularization was 93.8%. Three failures included 1 vasospasm, 1 distal embolus, and 1 side-branch occlusion. The total area of retrieved particles per basket was 2.76+/-6.49 mm(2) (range 0.0-40.3). Particles with a major axis >1 and >3 mm were detected in 29 (58.0%) and 6 (12.0%), respectively, of the examined filters. Collected particles consisted primarily of platelets and fibrin conglomerates, trapped erythrocytes, inflammatory cells, and extracellular matrix. Increased lesion length, increased reference vessel diameter, acute thromboses, and total occlusions were positively correlated with higher amounts of captured particles (p<0.05). Multivariate analysis incriminated declotting procedures as the only independent predictor of increased embolic burden (p<0.05). CONCLUSION: The embolism phenomenon during infra-aortic interventions is frequent and underestimated. The liberated particles consisted primarily of atheromatous plaque elements and thrombus. The reported data might support the application of a protective filter basket in selected subsets of lesions with a riskier embolic profile and whenever declotting procedures are performed.

Aged↗

Outflow protection filters during percutaneous recanalization of lower extremities' arterial occlusions: a pilot study.

PURPOSE: Filter devices are already employed for the protection of carotid, coronary and renal distal vascular bed during endovascular procedures. This is a pilot study investigating their feasibility, safety and distal emboli protection capability during recanalization of lower extremities' acute and subacute occlusions. MATERIALS AND METHODS: Study population included 16 patients, 11 with a subacute arterial occlusion and 5 with an acute episode. The Trap filter (Microvena, USA) and its successor the Spider filter (EV3, USA) were utilized. Subacute occlusions were dealt with standard angioplasty and stenting procedures, while acute ones were managed primarily with Angiojet rheolytic thrombectomy. Outflow arterial tree was checked angiographically in-between consequent procedural steps. Embolic material collected after filter recovery was analyzed histopathologically. Patients' follow-up was scheduled at 1 month. RESULTS: Seventeen filter baskets were applied in the recanalization of 16 target lesions in total. Mean length of the occluded segments was 6.1 (range: 2-15 cm; S.D. = 3.7 cm). Mean in situ time of the filters was 38.75 min (range: 20-60 min; S.D. = 12.71 min). Technical success rate of deployment and utilization of the filtration devices was 100% (17/17). Procedural success rate of the recanalization was 100% (16/16) without any clinical or angiographic evidence of periprocedural distal embolization. Macroscopic particulate debris was extracted from all the filters (17/17) containing fresh thrombus, calcification minerals, cholesterol and fibrin. Mean diameter of the largest particle per specimen was 1702.80 (range: 373.20-4680.00 microm; S.D. = 1155.12 microm). No adverse clinical events occurred at 1-month follow-up with 100% limb salvage (16/16). CONCLUSION: The application of outflow protection filters is safe, feasible and efficacious in hindering distal embolization complications and safeguarding the distal capillary bed. Nevertheless, this is a pilot study in a limited group. Further studies have to be contacted in order to provide evidence for a more general use of these devices.

Angiography, Digital Subtraction↗

The potential role of TGFbeta1, TGFbeta2 and TGFbeta3 protein expression in colorectal carcinomas. Correlation with classic histopathologic factors and patient survival.

PURPOSE: This study investigates the expression of tumor growth factors TGFbeta1, TGFbeta2 and TGFbeta3 in tissue material from patients with colorectal carcinoma and evaluates their correlation with known prognostic markers and patient survival. PATIENTS AND METHODS: The study included 124 patients with colorectal carcinoma. According to the TNM classification of malignant tumors, 26 tumors were identified as being stage I, 30 stage II, 48 stage III, and 20 stage IV, whereas 106 tumors were low-grade and 18 high-grade malignancies. On paraffin sections, the streptavidin-biotin technique using antibodies against TGFbeta1, TGFbeta2 and TGFbeta3 was applied. Morphological and immunohistochemical results were correlated with clinicopathologic parameters. RESULTS: TGFbeta1 protein was expressed in 88 out of 124 (71%) carcinomas, whereas TGFbeta2 and TGFbeta3 proteins were detected in all tumors examined. Normal colonic mucosal epithelial cells expressed TGFbeta2 (significantly less as compared to neoplastic cells; p < 0.01) and TGFbeta3 (p > 0.05 compared to neoplastic cells), but not TGFbeta1. Statistical analysis revealed a higher expression of TGFbeta1 in low-grade carcinomas (p = 0.009) and a higher presence of TGFbeta2 in advanced tumors (p = 0.008). TGFbeta1 expression was related with increased disease-free and overall survival (p < 0.05 each). The presence of TGFbeta2 was correlated with worse prognosis (p < 0.05). Cox analysis revealed that besides tumor grade and stage, TGFbeta1 expression constituted an independent prognostic factor. CONCLUSION: This study shows that in adenocarcinomas of the colon, there is a differential expression of TGFbeta1, TGFbeta2 and TGF3. TGFbeta1 may be implicated in the pathogenesis of these tumors, since it is expressed only in neoplastic but not in normal cells. TGFbeta1 is related with an increased disease-free and overall survival and constitutes an independent prognostic factor. In advanced stages, TGFbeta2 seems to be involved in tumor progression and is related with worse prognosis.

Adenocarcinoma↗

Computer-based malignancy grading of astrocytomas employing a support vector machine classifier, the WHO grading system and the regular hematoxylin-eosin diagnostic staining procedure.

OBJECTIVE: To investigate and develop an automated technique for astrocytoma malignancy grading compatible with the clinical routine. STUDY DESIGN: One hundred forty biopsies of astrocytomas were collected from 2 hospitals. The degree of tumor malignancy was defined as low or high according to the World Health Organization grading system. From each biopsy, images were digitized and segmented to isolate nuclei from background tissue. Morphologic and textural nuclear features were quantified to encode tumor malignancy. Each case was represented by a 40-dimensional feature vector. An exhaustive search procedure in feature space was utilized to determine the best feature combination that resulted in the smallest classification error. Low and high grade tumors were discriminated using support vector machines (SVMs). To evaluate the system performance, all available data were split randomly into training and test sets. RESULTS: The best vector combination consisted of 3 textural and 2 morphologic features. Low and high grade cases were discriminated with an accuracy of 90.7% and 88.9%, respectively, using an SVM classifier with polynomial kernel of degree 2. CONCLUSION: The proposed methodology was based on standards that are common in daily clinical practice and might be used in parallel with conventional grading as a second-opinion tool to reduce subjectivity in the classification of astrocytomas.

Artificial Intelligence↗

Automated segmentation of routinely hematoxylin-eosin-stained microscopic images by combining support vector machine clustering and active contour models.

OBJECTIVE: To develop a method for the automated segmentation of images of routinely hematoxylin-eosin (H-E)-stained microscopic sections to guarantee correct results in computer-assisted microscopy. STUDY DESIGN: Clinical material was composed 50 H-E-stained biopsies of astrocytomas and 50 H-E-stained biopsies of urinary bladder cancer. The basic idea was to use a support vector machine clustering (SVMC) algorithm to provide gross segmentation of regions holding nuclei and subsequently to refine nuclear boundary detection with active contours. The initialization coordinates of the active contour model were defined using a SVMC pixel-based classification algorithm that discriminated nuclear regions from the surrounding tissue. Starting from the boundaries of these regions, the snake fired and propagated until converging to nuclear boundaries. RESULTS: The method was validated for 2 different types of H-E-stained images. Results were evaluated by 2 histopathologists. On average, 94% of nuclei were correctly delineated. CONCLUSION: The proposed algorithm could be of value in computer-based systems for automated interpretation of microscopic images.

Algorithms↗

BCL-2 and BAX expression and cell proliferation, after partial hepatectomy with and without ischemia, on cholestatic liver in rats: an experimental study.

BACKGROUND: Liver regeneration after partial hepatectomy (PHx) is regulated by several factors that activate or inhibit hepatocyte proliferation. Apoptosis seems to play an important role in cellular proliferation and liver regeneration. This study investigates the expression apoptosis-associated genes bcl-2 and bax, and the presence of apoptosis and cell proliferation after PHx, in normal and jaundiced rats with or without superimposed ischemia. MATERIALS AND METHODS: The study included 50 male Wistar rats assigned into; five groups (10 rats each). On day 0, rats of groups II, IV, and V underwent common bile duct ligation (BDL). On day 10, total liver ischemia (TLI) (occlusion of hepatic artery and portal vein-TLI) for 30 min was performed on animals of group V. When TLI was completed, all 30 animals (of groups I, IV, and V) underwent PHx (68%). Animals of group III underwent only TLI for 30 min. Rats of groups I, IV, and V were sacrificed 24 and 48 h after PHx was completed. Rats of group II were sacrificed 10, 11, and 12 days after BDL. Rats of group III were sacrificed immediately, 24 and 48 h after TLI completion. Liver tissue was obtained and pathologic examination included: (a) H&E stain, (b) in situ hybridization (detection of bcl-2 and bax mRNA) in paraffin sections, (c) Western blot analysis for the evaluation of bcl-2 and bax protein levels, (d) in situ hybridization (TUNEL) for the detection of apoptotic bodies, and (e) immunohistochemical stains (streptavidin-biotin method) in paraffin sections to detect cells that (i) express bcl-2 and bax proteins and (ii) undergo proliferation (Ki67+ cells). Results were expressed following morphometric analysis. RESULTS: Before hepatectomy, bcl-2 (protein or mRNA) levels were higher in jaundiced rats vs controls. Furthermore, bax (protein or mRNA) levels and apoptotic body index (ABI) were higher in cholestatic livers. After hepatectomy, there was an early decrease in the protein and mRNA levels of antiapoptotic gene bcl-2 and a late increase of proapoptotic gene bax and the ABI, compared to controls. Cell proliferation of hepatocytes was lower in group V (BDL + TLI) compared to that of groups II and IV (BDL). CONCLUSIONS: This study shows that apoptosis takes place in cholestatic livers with or without superimposed ischemia and may contribute in the impaired regenerative response observed in livers of jaundiced rats after partial hepatectomy.

Animals↗

Potential role of bcl-2 and bax mRNA and protein expression in chronic hepatitis type B and C: a clinicopathologic study.

Bcl-2 oncoprotein regulates programmed cell death by providing a survival advantage to rapidly proliferating cells, and bax protein promotes apoptosis by enchanting cell susceptibility to apoptotic stimuli. In this study, we assessed the expression of bcl-2 and bax in liver biopsies from patients with chronic hepatitis (CH) Type B (HBV) and C (HCV). The study comprised 65 liver biopsies from 65 patients with HBV (n = 37) and HCV (n = 28) and 10 normal liver biopsies as controls. The HAI score ranged from 3/18-13/18, and the fibrosis Stage, from 1-6 (7 HBV/10 HCV). Pathologic examination included the following: (1) immunohistochemical stains in paraffin sections for bcl-2 and bax protein expression, (2) Western blot analysis (bcl-2 and bax protein levels evaluation), (3) ISH (detection of bcl-2 and bax mRNA), and (4) ISH (TUNEL-ABI [apoptotic body index]). In CH cases, both bcl-2 and bax protein and mRNA were detected in portal and intralobular lymphocytes and in cholangiolar epithelial cells in interface areas and fibrous bands. Bax protein and mRNA was expressed within hepatocytes and epithelial cells of interlobular ducts in portal tracts. Bcl-2 mRNA was present in periportal hepatocytes only in cases with Stage 5-6 fibrosis. Western blot analysis showed a decreased bcl-2 and an increased bax expression toward advanced fibrotic stages. In CH cases, ABI was reverse correlated with the percentage of bcl-2 expression and was correlated directly with the percentage of bax expression (P <.001). The results of this study suggest that in cases of chronic HBV or HCV infection, bax may be involved in the hepatocyte cycle regulation during infection, whereas its expression in intraportal bile duct epithelium implies that this protein enhances susceptibility of these particular cells to apoptosis. The increased bax expression and ABI in fibrosis Stages 1-5, imply that they are responsible for hepatocytes depletion through apoptosis, during progress of liver fibrosis and fibrous tissue accumulation, until cirrhosis is established. Bcl-2 mRNA expression in periportal hepatocytes only in Stages 5 and 6 suggests that this oncogene is involved in the late stages of progressive liver fibrosis and failure and furthermore that periportal hepatocytes are resistant to apoptosis. Bcl-2 expression, in cholangioles of interface area, suggests that this oncoprotein may be involved in growth regulation of these epithelial cells. Further research is warranted to specify the exact role of apoptosis and apoptotic genes involved in liver fibrosis process in cases of chronic HBV and HCV infection. This may lead to new strategies in the management of human liver disease to prevent the progression to chronic liver failure.

Adult↗

Non-palpable breast carcinomas: correlation of mammographically detected malignant-appearing microcalcifications and molecular prognostic factors.

Screening mammography has greatly increased the number of non-palpable breast carcinomas diagnosed in asymptomatic women. Malignant-appearing microcalcifications represent one of the earliest mammographic findings of non-palpable breast carcinomas. Many studies have attempted to correlate radiological and histological features of malignant-appearing microcalcifications. In the present study, we evaluated the association between mammographically detected malignant-appearing microcalcifications and the expression profile of selected biological markers in non-palpable breast carcinomas. Two hundred and eighty patients with non-palpable suspicious breast lesions that were detected during screening mammography were studied. All patients underwent mammographically-guided needle localization-excision breast biopsy. Histological examination showed 74 (26.4%) carcinomas of various subtypes. Immunohistochemistry was carried out in 58/74 carcinomas by using a panel of monoclonal and polyclonal antibodies against estrogen receptor (ER), progesterone receptor (PR), HER-2/neu, Bcl-2, Bax, Fas and DNA fragmentation factor (DFF). Malignant-appearing microcalcifications was the major mammographic finding in 45/58 (77%) patients. Nuclear ER positivity (65.5%) and PR positivity (46.5%) of non-palpable breast carcinomas were statistically correlated with malignant-appearing microcalcifications (p < 0.01 and p < 0.05, respectively). Statistically significant associations were also found between malignant-appearing microcalcifications and HER-2/neu positivity (p < 0.01), Bax positivity (p < 0.01), Fas positivity (p < 0.05) and DFF positivity (p < 0.01), whereas no statistical correlation was found with Bcl-2 positivity (p > 0.05). Malignant-appearing microcalcifications detected during screening mammography represent a diagnostic, prognostic and therapeutic challenge. The mammographic/biological associations and their potential implications in the management of women with non-palpable breast carcinomas are thoroughly discussed.

Adult↗

Survival for 5 years after repeat liver resections and multimodality treatment for metastatic intestinal leiomyosarcoma: report of a case.

The liver is a common site of metastases from gastrointestinal or retroperitoneal leiomyosarcomas. Although repeat liver resections can prolong survival, to our knowledge, 5-year survival has never been achieved. We report the case of a woman who has survived for 5 years since her first liver resection, during which time five more resections have been performed, in combination with systemic chemotherapy and radiofrequency ablation.

Adult↗

Expresssion of bcl-2 oncoprotein in cases of acute and chronic viral hepatitis type B and type C: a clinicopathologic study.

The bcl-2 gene is involved in the regulation of programmed cell death by providing a survival advantage to rapidly proliferating cells. This study investigates bcl-2 protein expression in liver biopsies with acute or chronic viral hepatitis B (HBV) or C(HCV). The study comprised 60 liver biopsies with hepatitis B or C. These included 10 biopsies from cases with acute lobular hepatitis (ALH), and 50 with chronic hepatitis (CH). In addition, 10 liver biopsies were used as controls. In CH cases, HAI grade ranged from 3/18 to 15/18, and stage from 1 to 6 (6HBV/8HCV). Immunohistochemical bcl-2 expression was assessed using the streptavidin-biotin method and the presence of apoptotic cells was evaluated by TUNEL method. Immunohistochemical and in situ hybridization (TUNEL) results were expressed following morphometric analysis. In CH cases, bcl-2 was detected in portal and intralobular lymphocytes, and in cholangiolar epithelial cells of the interface area. In ALH and control cases, bcl-2 was expressed only in lymphocytes. Lymphocytic bcl-2 expression was correlated directly with categories A, C and D of HAI (P < 0.001), whereas the degree of fibrosis was reversibly correlated to bcl-2 expression. In conclusion, in cases of chronic hepatitis, bcl-2 expression in cholangioles of interface area suggests that this oncoprotein may be involved in regulation of growth of these epithelial cells.

Adult↗

Neural network-based segmentation and classification system for automated grading of histologic sections of bladder carcinoma.

OBJECTIVE: To develop an image analysis system for automated nuclear segmentation and classification of histologic bladder sections employing quantitative nuclear features. STUDY DESIGN: Ninety-two cases were classified into three classes by experienced pathologists according to the WHO grading system: 18 cases as grade 1, 45 as grade 2, and 29 as grade 3. Nuclear segmentation was performed by means of an artificial neural network (ANN)-based pixel classification algorithm, and each case was represented by 36 nuclei features. Automated grading of bladder tumor histologic sections was performed by an ANN classifier implemented in a two-stage hierarchic tree. RESULTS: On average, 95% of the nuclei were correctly detected. At the first stage of the hierarchic tree, classifier performance in discriminating between cases of grade 1 and 2 and cases of grade 3 was 89%. At the second stage, 79% of grade 1 cases were correctly distinguished from grade 2 cases. CONCLUSION: The proposed image analysis system provides the means to reduce subjectivity in grading bladder tumors and may contribute to more accurate diagnosis and prognosis since it relies on nuclear features, the value of which has been confirmed.

Algorithms↗

Acute lymphoblastic leukemia relapse in the breast diagnosed with gray-scale and color Doppler sonography.

Extramedullary relapses of acute lymphoblastic leukemia (ALL) in children and young adults are rare and in most cases are localized in the central nervous system, testes, or both. We describe a rare case of extramedullary relapse of ALL in the breast of a 17-year-old girl. The patient, who had been diagnosed with ALL 1 year before and had been in complete remission for 5 months, was admitted to the hospital for investigation of a mass in her left breast. On clinical examination, she had a large, palpable, nontender mass in her left breast. Mammography revealed a very dense mass. On gray-scale and power Doppler sonography, the appearance of the mass was consistent with malignancy. Histopathologic examination of a surgical biopsy specimen permitted accurate diagnosis of ALL.

Adolescent↗

Automated diagnosis of brain tumours astrocytomas using probabilistic neural network clustering and support vector machines.

A computer-aided diagnosis system was developed for assisting brain astrocytomas malignancy grading. Microscopy images from 140 astrocytic biopsies were digitized and cell nuclei were automatically segmented using a Probabilistic Neural Network pixel-based clustering algorithm. A decision tree classification scheme was constructed to discriminate low, intermediate and high-grade tumours by analyzing nuclear features extracted from segmented nuclei with a Support Vector Machine classifier. Nuclei were segmented with an average accuracy of 86.5%. Low, intermediate, and high-grade tumours were identified with 95%, 88.3%, and 91% accuracies respectively. The proposed algorithm could be used as a second opinion tool for the histopathologists.

Algorithms↗

The potential role of bcl-2 mRNA and protein exression in hepatocellular carcinomas.

BACKGROUND: The bcl-2 gene regulates programmed cell death by providing a survival advantage to rapidly proliferating cells. In several malignant tumors bcl-2 presence has been associated with favorable prognosis. In the liver, bcl-2 is expressed in bile ductular cells and tumors of biliary origin. This study investigates the expression of bcl-2 mRNA and protein in hepatocellular carcinomas (HCC). MATERIALS AND METHODS: The study comprised 35 primary HCC resected from 35 patients; 21 males and 14 females, aged from 43-59 years. The tumors were graded according to Edmondson criteria. In 28 cases HCC was developed on the background of cirrhotic liver. In 9 instances the patients received chemoembolization before surgery. The presence of bcl-2 mRNA was assessed on paraffin-embedded, 4 microm-thick sections, using a standard in situ hybridization method with a commercially available bcl-2 probe (Maxim Biotech, USA), while the streptavidin-biotin immunohistochemical technique was employed to detect bcl-2 protein expression using the monoclonal anti-bcl-2 antibody (DAKO, USA). The results were expressed as % of tumor-positive cells following morphometric analysis. RESULTS: The in situ hybridization study revealed bcl-2 mRNA expression in 25 out of 35 (70%) HCC. In addition, bcl-2 mRNA was detected in hyperplastic cholangioles within fibrous septae in the periphery of the tumors. Immunohistochemical staining failed to reveal any bcl-2 protein expression in tumor cells of HCC, whereas hyperplastic cholangioles of the fibrous septae, in the periphery of the tumors and intratumoral lymphocytes displayed a strong-positive cytoplasmic (perinuclear) stain. No significant correlations were recorded between bcl-2 mRNA expression and tumor histological pattern, grade, stage and the use of previous chemoembolization. CONCLUSION: In hepatocellular carcinomas, the bcl-2 gene is frequently present but its protein product is absent. This suggests a post-translational mechanism of bcl-2 protein degradation, indicating that bcl-2 does not play a substantial role in the progress of hepatocellular carcinoma.

Adult↗