Tissue expression of VEGF in cervical intraepithelial neoplasia and cervical cancer.

PURPOSE: To examine the expression of vascular endothelial growth factor (VEGF) in the cervical tissue of individuals divided into the control group (normal cervix), group A (HSIL lesions), and group B (cervical cancer, FIGO stage I-IIA). Analyzed was also the expression of VEGF between groups and subgroups in group A and B. The expression of VEGF was also compared with histopathological parameters in group B. METHODS: Examined was the histopathological material taken from 109 operated patients. The patients were divided into 3 groups based on the definitive histopathological findings: control group (30 patients), group A (33 patients), and group B (46 patients). Immunohistochemistry was performed to examine the expression of VEGF. RESULTS: The expression of VEGF was negative in the control group, while in 11 patients (33.33%) from group A and 28 patients (60.87%) from group B it was significantly different (p<0.05) compared to the control group. There was neither statistically significant difference in the expression of VEGF in group A regarding the type of intraepithelial lesion, nor in group B regarding the FIGO disease stage (p>0.05). In patients with poor histopathological prognostic parameters such as tumor diameter ≥ 2 cm (24/46), depth of stromal invasion ≥ 10 mm (32/46), positive lymph nodes (17/46), and with infiltration of the uterine body (11/46) a statistically significant difference was confirmed regarding the expression of VEGF. CONCLUSION: The increased VEGF expression in groups A and B compared with the control group indicated the importance of VEGF as a proangiogenic factor in neoangiogenesis in precancerous and cancerous changes in the cervix. The frequent expression of VEGF in the subgroup of patients with poor histopathological prognostic factors (group B) indicated the importance of the activity of proangiogenic factors in the process of cervical cancer neoangiogenesis. Further investigations should be aimed at these markers as prognostic factors in the high risk group of patients with cervical cancer who should receive adjuvant therapy after radical operation and consider using antiangiogenic drugs as part of adjuvant treatment.

Cyclooxygenase-2 expression in cervical cancer.

BACKGROUND/AIM: Cyclooxygenase (COX) or prostaglandin H2 synthase is the first enzyme that catalyzes the first two steps in the biosynthesis of prostaglandins from arachidonic acid. The aim of the study was to determine the expression level of COX-2 in patients with cervical cancer and compare it with that in the control group with no cervical pathology. METHODS: The study included 76 patients divided into two groups: the control group--30 patients without histopathological changes and the group A--46 patients with cervical cancer, FIGO stage IB-IIA. Histopathological and immunohistochemical analyses were performed in these two groups of patients. RESULTS: In the control group, the expression of COX-2 was not confirmed compared to the group A of 26 (56.52%) patients. The expression of COX-2 showed a statistically significant difference in the presence oflymphocytic stromal infiltration (p = 0.0053). The expression of COX-2 was more pronounced in the stromal tissue without lymphocytic infiltration (80% vs. 20%). CONCLUSION: A higher expression of COX-2 in cervical carcinoma without stromal lymphocytic infiltration suggests a possible paradoxical effect of COX-2 in immunosuppression. Frequent COX-2 expression in the subgroup with poor prognostic histological parameters in the group A indicates the importance of COX-2 expression in the carcinogenesis of cervical cancer.

[Histochemical and immunohistochemical analysis of ruptured atherosclerotic abdominal aortic aneurysm wall].

BACKGROUND/AIM: The main complication of the atherosclerotic abdominal aortic aneurysm (AAA) is her rupture that begins with lesion in intima and rupture. The purpose of this work was to determine immunocytochemical and morphofunctional characteristics of the cells in aortic wall in ruptured atherosclerotic abdominal aortic aneurysm. METHOD: During the course of this study, 20 samples of atherosclerotic AAA were analyzed, all of them obtained during authopsy. The samples were fixed in 4% formalin and embedded in paraffin. Sections of 5 microm thickness were stained histochemically (of Heidenhain azan stain and Periodic acid Schiff--PAS stain) and immunocytochemically using a DAKO LSAB+/HRP technique to identify alpha-smooth muscle actin (alpha-SMA), vimentin, myosin heavy chains (MHC), desmin, S-100 protein, CD45 and CD68 (DAKO specification). RESULTS: The results of our study showed that ruptured atherosclerotic AAA is characterized by a complete absence of endothelial cells, the disruption of basal membrane and internal elastic lamina, as well as a presence of the remains of hypocellular complicated atherosclerotic lesion in intima. On the plaque margins, as well as in the media, smooth muscle cells (SMCs) are present, which express a alpha-SMA and vimentin (but without MHC or desmin expression), as well as leukocyte infiltration, and a large number of foam cells. Some of the foam cells show a CD68- immunoreactivity, while the others show vimentin- and S-100 protein-immunoreactivity. Media is thinned out with a disorganized elastic lamellas, while adventitia is characterized by inflammatory inflitrate (infection). CONCLUSION: Rupture of aneurysm occurs from the primary intimal disruption, which spreads into thinned out media and adventitia. Rupture is caused by unstable atherom, hypocellularity, loss of contractile characteristics of smooth muscle cells in intima and media, neovascularization of the media, as well as by the activity of the macrophages in the lesion.

[Immunocytochemical characteristics of submucosal uterine myomas].

BACKGROUND/AIM: Myomas of the uterus, the most common benign tumors, have been studied for decades from the aspects of different basic and clinical disciplines. Despite this fact, their pathogenesis is still poorly understood. The aim of this study was to determine immunocytochemical characteristics of smooth muscle cells and connective tissue components of submucosal myomas of the uterus. METHOD: During the course of this study, 25 samples of submucosal myomas of the uterus were analyzed, all of them obtained during the surgery, after abdominal histerctomy by Aldridge. The samples were fixed in 4% formalin and embedded in paraffin. Sections of 5 microm thickness were stained immunocytochemically using the DAKO LSAB+/HRP technique to identify alpha-smooth muscle actin (alpha-SMA), vimentin, desmin, CD34, CD45, CD68 and PCNA (DAKO specification). RESULTS: Our results suggest that submucosal myomas of the uterus are build-up of smooth muscle cells which are immunoreactive to alpha-SMA and desmin, but also to a certain number of smooth muscle cells which are immunoreactive to alpha-SMA and vimentin. Some of vimentin-immunoreactive cells also show an immunoreactivity of PCNA. In the build-up of connective stroma CD34-immunoreactive fibroblasts and neovascular formations are also present. By examining the distribution of CD45 antigen, at all the analyzed samples we observed a weak reaction. CONCLUSION: Submucosal myomas of the uterus are made-up of smooth muscle cells of the highly differentiated contractile phenotype (alpha-SMA- and desmin-immunoreactivity), as well as smooth muscle cell of the synthetic phenotype which proliferate (alpha-SMA-, vimentin- and PCNA-immunoreactivity). In submucosal myoma of the uterus there is a significant presence of connective tissue as a result of synthetic activity of fibroblasts, which clearly differ in their immunocytochemical characteristics from smooth muscle cells of the synthetic phenotype.

Comethylation of p16 and MGMT genes in colorectal carcinoma: correlation with clinicopathological features and prognostic value.

AIM: To investigate the significance of p16 and O(6)-methylguanine-DNA methyltransferase (MGMT) genes promoter hypermethylation and K-ras mutations on colorectal tumorigenesis and progression. METHODS: p16 and MGMT methylation status was examined on 47 tumor samples, and K-ras mutational status was examined on 85 tumor samples. For methylation analysis, a methylation specific PCR (MS-PCR) method was used. RESULTS: p16 and MGMT promoter methylation was found in 51% (24/47) and 43% (20/47) of CRCs, respectively, and the K-ras mutation was found in 44% (37/85) of CRCs. Comethylation of p16 and MGMT genes was significantly associated with lower aggressiveness of the disease within a two-year period of observation. Only 27% of patients with simultaneous p16 and MGMT methylation showed the detectable occurrence of metastasis and/or death, compared to 67% of patients without double methylation or with no methylation (3/11 vs 22/33, P < 0.05, chi(2)-test). In addition, p16 and MGMT comethylation showed a trend toward an association with longer survival in patients with CRCs (35.5 +/- 6.0 mo vs 23.1 +/- 3.2 mo, P = 0.072, Log-rank test). Progression of the disease within a two-year period was observed in 66% of patients carrying the K-ras mutation, compared to only 19% of patients with wild type K-ras (29/44 vs 7/37, P < 0.001, chi(2)-test). The presence of the K-ras mutation significantly correlated to shortened overall survival (20.0 +/- 1.9 mo vs 37.0 +/- 1.8 mo, P < 0.001, Log-rank test). The comethylation of p16 and MGMT genes was significantly associated with lower aggressiveness of the disease even when K-ras mutations were included in the analysis as an independent variable. CONCLUSION: Our data suggest that comethylation of promoters of p16 and MGMT genes could have a prognostic value in patients with CRC. Specifically, concurrent methylation of both genes correlates with better prognosis.

[Detection of free cancer cells in peritoneal cavity in patients surgically treated for gastric adenocarcinoma].

BACKGROUND/AIM: Peritoneal metastasis is a leading cause of therapeutic failure after an operative treatment of patients with gastric adenocarcinoma. Free cancer cells might induce or indicate an early peritoneal seeding with a subsequent peritoneal metastasis. The aim of this study was to determine the frequency of the presence of free cancer cells in the peritoneal cavity in the patients surgically treated for gastric adenocarcinoma, and its relation to certain clinical, operative and pathohistological paramethers. METHODS: Inside a period from April 2000, and April 2004, the total of 100 patients underwent intraoperative peritoneal lavage for cytological examination. Immediately after the laparotomy, 200 ml physiologic saline, heated to 37 degrees C, was introduced into the abdominal cavity, mannualy dispersed and collected from the region around the gastric tumor and the pouch of Douglas. The nucleated cell layer was smeared on four glass slides for every patient and dyed with May-Grünwald-Giemsa stain. The cytological findings were defined as positive or negative according to the presence of cancer cells. The frequency of positive cytological findings was compared to the location and the diameter of the cancer, pathohistological type of carcinoma, pathohistological stage of the disease, lymph node and the liver and/or peritoneal metastases and the type of surgical procedure. RESULTS: Free cancer cells were found in 24 (24%) of the patients, while in 76 (76%) of them cytological findings were negative. A statistically highly significant difference (p < or = 0.001) in the frequency of positive cytological finding was found between the groups of patients with and without cancer invasion of serosa, with cancer diameters > 5 cm and < or = 5 cm, in the stage of disease I, II and III, IV, with macroscopically present and without metastases, with re section and D2 lymphadenectomy and palliative procedure. Free cancer cells were statistically more frequently (p < or = 0.05) detected in the patients with lymph nodes metastases comparing to the patients with out lymph nodes involvement. The results of the univariate analysis showed that the cancer diameter > 5 cm, tumor invasion of serosa, pathohistological stage of the disease III and IV and macroscopically visible metastases were the most important risk factors for the free cancer cells detection. CONCLUSION: Peritoneal lavage cytology was shown to be a useful tool for the detection of the group of patients with greatest risk of peritoneal dissemination. The frequency of positive cytological findings was highly associated with the diameter of the tumor and the cancer invasion of serosa. Cytological examination of peritoneal lavage fluid improved the accuracy of staging and selection of patients who might have benefit from neoadjuvant chemotherapy.

[Early complications following radical surgical treatment of patients with gastric adenocarcinoma].

BACKGROUND/AIM: Surgical treatment of patients with gastric adenocarcinoma means the total excision of a tumor and the pathways of its spreading with the risk of operational complications as low as possible. The aim of this study was to evaluate the type and frequency of early postoperative complications and mortality after a radical surgical treatment of patients with gastric adenocarcinoma. METHODS: Complication rates and postoperative mortality were studied in 70 consecutive patients in whom a radical surgical procedure, gastrectomy (total or subtotal) with D2 lymphadenectomy, was performed. In the early postoperative period, the frequencies of general and specific complications were detected. The frequencies of complications were compared between the groups of patients according to the defined clinical, operative and pathohistological paramethers. RESULTS: The overall morbidity and mortality rates were 27.14% and 5.71%, respectively Pancreatic fistula in five, and pleural effusion in three patients were the most frequently registered complications. Three of four deaths occured in patients older than 70 years, with the stage III and IV of the disease, and in all of them total gastrectomy with splenectomy was performed. A statistically significant difference (p < 0.05) in complication rates was found between the groups of patients with and without splenectomy and with the tumors > 5 cm and < or = 5 cm. CONCLUSION: Radical surgical treatment of patients with gastric adenocarcinoma might be done with an acceptable morbidity and mortality if it is performed by the surgeons with the experience in D2 lymphadenectomy technique. A diameter of the tumor > 5 cm, and splenectomy, and/or splenopancreatectomy are the most important risk factors for the occurrence of complications and modifications of D2 lymphadenectomy technique with limited indications for splenic and/or pancreas resection can improve treatment results. An individual approach and the appropriate selection of the surgical procedure are necessary in patients older than 70 years.