Autonomic dysfunction of gastric motility in major depression.

BACKGROUND: Patients suffering from major depressive disorder (MDD) often complain about somatic symptoms. Cardiac complaints have been examined predominantly. However, gastrointestinal complaints are also reported frequently and are associated with worse outcomes. The research concerning changes in gastric motility of these patients is rather sparse. The aim of our study was to determine dysfunction of gastric motility and gastrointestinal symptoms in MDD. The duration and severity of MDD were examined regarding its influence over gastric emptying. METHODS: Gastric emptying was determined by a 13C-acetate breath test in patients with MDD (n = 29) and healthy control subjects (n = 51). Prior to this, depressive illness was operationalized using external and self-assessment scales (HAMD-21, MADRS, BDI, CGI). Whether the severity or duration of MDD influenced the gastric emptying parameters was examined using Spearman's correlation. In addition, autonomic complaints were recorded by means of an ANS score. Each ANS score item was determined using a Mann-Whitney U or Kruskal-Wallis test concerning the gastric emptying parameters. RESULTS: There was a significant difference in the parameters of the maximum gastric emptying rate (Tmax) and gastric half emptying time T1/2b between patients with MDD and healthy control subjects (Tmax 66.21min vs 53.35min, p < 0.006, T1/2b 207.59min vs 133.27min, p < 0.005). There was a significant negative correlation between Tmax and the severity of MDD determined with the depression rating scales BDI (Spearman's rank - 0.521, p = 0.013) and HAMD-21 (r - 0.384, p = 0.048). No correlation was found between the duration of MDD and the maximum gastric emptying rate (r - 0.125, p = 0.519) and gastric half emptying time (r - 0.62, p = 0.749). CONCLUSION: Gastrointestinal motility is significantly impaired in patients with MDD compared to healthy control subjects. Autonomic complaints were indicated frequently in MDD patients. The duration of MDD had no influence over the time of gastric emptying. There was a significant negative correlation between the severity of MDD and Tmax, indicating that the Tmax was reached earlier with the progression of MDD. The slowing of gastric motility in MDD patients is likely a result of a dysfunction of the autonomic nervous system.

Relations between the expression of p53, c-erbB-2, Ki-67 and HPV infection in cervical carcinomas and cervical dysplasias.

BACKGROUND: The analysis of mutual relations between HPV infection and the expression of cancer gene products and proliferative activity in cervical carcinomas and dysplasias. MATERIALS AND METHODS: The expression of p53, c-erbB-2 oncoproteins and proliferative activity (Ki-67) was evaluated immunohistochemically in 41 cervical carcinomas and 29 dysplasias. HPV infection (type 16, 18) was assessed by in situ hybridization technique. RESULTS: HPV-positive carcinomas were found in 68.3% of cases. HPV type 16 infection were detected in 54% and HPV 18 in 39% of carcinomas. Simultaneous appearance of both virus types was shown in 25% of carcinomas. In dysplastic lesions, HPV infection was observed in 62.1% of cases. HPV type 16 was found in 34.5% and HPV 18 in 44.8% of patients. Both virus types were found in 17.2% of dysplasias. HPV infection was more extensive in cervical carcinomas than in dysplasias. Similarly the expression of oncoproteins was more intensive and referred to a higher percentage of cells in carcinomas. No relations between p53, c-erbB-2 overexpression and HPV infection were found. Ki-67 activity was found in a higher percentage of HPV-positive than in HPV-negative, both carcinomas and dysplasias. CONCLUSIONS: HPV infection, especially accompanied by increase of proliferative activity in dysplasias may define the cell subpopulation predisposed to malignant process development. The employment of in situ hybridization technique appears to be useful in detecting the viral infection in cytological smears even with no morphological changes in the cells.

Expression and mutation of p53 in tumor effusion cells of patients with ovarian carcinoma: response to cisplatin-based chemotherapy.

p53 alterations are considered as one of the most important factors responsible for drug resistance in ovarian carcinomas, although the relationship between p53 gene status and response to cisplatin-based chemotherapy in ovarian cancer patients remains unclear. The aim of the study was to evaluate the relationship between p53 protein accumulation, p53 gene mutation and response to cisplatin-based chemotherapy in patients with ovarian carcinoma considering conventional clinicopathological parameters. Tissue sections and corresponding cyst and/or ascitic fluid cells from 79 patients with epithelial ovarian cancer were analyzed immunohistochemically for p53 expression. The PCR-SSCP analysis was performed in 25 cases and the results were compared with immunohistochemical data. It was demonstrated that p53 expression reaching approximately 50% of positive cells in immunostaining was usually associated with PCR-amplified exons showing abnormal migration and suspected for mutation. p53 gene changes were not correlated with histological structure, grade of differentiation or residual tumor after cytoreductive surgery, despite being detected more frequently in III/IV than in II FIGO stages and in patients with residual disease above 2 cm. A significant correlation between p53 accumulation and p53 gene alteration and poor response to cisplatin-based chemotherapy was shown. The overall survival time of patients decreased with an increase in p53 protein expression. A strong p53 expression especially accompanied by p53 changes detectable by PCR-SSCP analysis appears to be a good indicator of the resistance to cisplatin-based chemotherapy. The association between strong p53 overexpression and shorter overall survival time was also revealed.

Morphological and phenotypic characterization of a new established ovarian carcinoma cell line (OvBH-1).

BACKGROUND: A limited number of permanent ovarian carcinoma cell lines have been described so far and the majority of them have been derived from ovarian ascitic fluid cells taken from patients with serous ovarian carcinoma usually after chemotherapy treatment. MATERIALS AND METHODS: The cells for culture were obtained from ascitic fluid cells of a patient with ovarian clear cell carcinoma. Cytomorphological analysis of cultured cells at early and late passages was performed by hematoxylin-eosin staining. Immunophenotypic characterization of cells was performed using the following monoclonal antibodies against: intermediated cellular filaments (CK 6/18, CK 7, CK 1,5,6,8,10,14,18, V9) ovarian carcinoma-associated antigens (OC125, OV-TL3), carcinoembryonic antigen, p53 and c-erbB-2 oncoproteins. RESULTS: In the established ovarian carcinoma cell line (OvBH-1) two morphologically distinct cell types were recognized. Cytomorphologically the dominating type appears to frankly malignant features. The second cell subtype showed a lower degree of malignant features. The epithelial origin of both cell types was confirmed by immunohistochemical staining using antibodies against different cytokeratin epitopes. The expression of tumor-associated antigens (CA125, OV-TL3) was found in both cell subtypes reflecting their origin from ovarian carcinoma. The cell line was negative for CEA staining. The genetic defects of cultured cells were revealed by detection of p53 and c-erbB-2 overexpression. The level of both oncoproteins and especially c-erbB-2 was higher in the cell subtype with frankly malignant morphological features. CONCLUSIONS: A new established, well characterized ovarian clear cell carcinoma line OvBH-1 provides an experimental model for further investigation of the biological alterations responsible for carcinogenesis and chemoresistance of this uncommon subtype of epithelial ovarian carcinomas.

Epidermal growth factor receptor and c-erbB-2 oncoproteins in tissue and tumor effusion cells of histopathologically different ovarian neoplasms.

The overexpression of two growth factor receptors - epidermal growth factor receptor (EGFR) and c-erbB-2 - was evaluated immunohistochemically in malignant and benign ovarian neoplasms, considering the stage of the disease and histology of tumors. The comparison of EGFR and c-erbB-2 reactivity in tissue sections and respective cyst and/or ascitic fluid cells was also performed. c-erbB-2 expression was detected in 44.4% of ovarian carcinomas, and in benign neoplasms there was no evidence of its staining, while EGFR reactivity was found both in malignant (58.7%) and benign (50%) tumors. Significant heterogeneity of staining was observed, however, the relationship between EGFR and c-erbB-2 expression in tissue sections and cyst and/or ascitic fluid cells in individual patients was evident. The expression of both growth factor receptors was not correlated with histopathological subtypes of ovarian neoplasms. The c-erbB-2 oncoprotein was detected more frequently in III/IV than in I/II stages according to the criteria of the International Federation of Gynecology and Obstetrics (FIGO) and the EGFR expression was independent of the clinical advancement of the disease. The coexpression of c-erbB-2 and EGFR was shown in 32% of ovarian carcinomas, and it dominated in cases with FIGO stages III/IV. Our results indicate that the increase of the EGFR expression appears to be associated with early stages of ovarian tumorigenesis, and the enhancement of c-erbB-2 reactivity may cooperate with EGFR activation in the development and progression of ovarian carcinomas.

p53 autoantibodies in the sera, cyst and ascitic fluids of patients with ovarian cancer.

The presence of p53 autoantibodies and p53 protein overexpression in ovarian carcinoma patients were determined and compared. p53 antibodies were detected in sera samples, cyst and/or ascitic fluids of individual patients by two separate techniques (ELISA assay and immunoblot). p53 protein accumulation was assessed immunohistochemically in tissue sections and corresponding tumor effusion cells. The relations between p53 overexpression, the presence of p53 autoantibodies and histology of tumors, grade of differentiation and clinical stage of the disease were considered. p53 expression was found in 20 of 46 (43.5%) ovarian carcinomas and significant relationship between p53 reactivity in tumor tissue and effusion cells in individual patients was evident. In the subset of carcinomas with detectable p53 accumulation only two cases (one serous, one endometrioid) were associated with the presence of p53 autoantibodies (10%). Among 26 p53- negative carcinomas also two cases (7.6%) were seropositive. The strong correlation between the presence of p53 autoantibodies in the sera and respective cyst or ascitic fluids were revealed with no exception of this coincidence. There was no association between the detection of antibodies against p53 and FIGO stage and tumor grade. Our results clearly indicate that p53 overexpression is not sufficient to elicit p53 humoral response in ovarian carcinoma patients. The presence of p53 autoantibodies in this type of cancer is not a frequent event and their importance as independent prognostic factor seems to be very limited.

Prospective studies of p53 and c-erbB-2 expression in relation to clinicopathological parameters of human ductal breast cancer in the second stage of clinical advancement.

BACKGROUND: The association between p53 and c-erbB-2 overexpression relation to ER status in ductal breast carcinoma is still unclear. Our aim was investigate the prognostic importance of the overexpression of c-erbB-2, p-53 factor, and ER status in stage II of human ductal breast cancer. MATERIALS AND METHODS: Th. expression of c-erbB-2 and p53 oncoproteins was evaluated by immunoperoxidase technique (PAP) in 62 cases of ductal breast carcinoma. The relationship between these cell growth regulatory factors was estimated and compared with the presence estrogen receptor (ER), tumor grading, tumor size, lymph node involvement, age patients and number of relapses up to the second year after surgery. RESULTS: c-erbB-2 overexpression was found in 44% and p53 in 45% of carcinomas. ER level was usually inversely proportional to the presence of studied molecular markers. Stratifying patients on the basis of c-erbB-2, p53 and ER status revealed that the combination c-erbB-2 and p53 overexpression accompanied by undetectable ER, identified the population of poorly differentiated tumors and patients with a high incidence of axillary lymph node metastases and shorter relapse time. On the other hand, undetectable values of molecular markers were associated with a low grade of tumors and a lack of lymph nodes involvement. CONCLUSIONS: Estimation of c-erbB-2, p53 and ER status seems to be a powerful tool to discriminate between different phenotypes of breast carcinoma. c-erbB-2 and p53 oncoproteins have been recognized as independent molecular markers of aggressive tumor behaviour.

p53, c-erbB-2 and p21ras expression in tumor effusion cells of patients with histopathologically different ovarian neoplasms.

The overexpression of p53, c-erbB-2 and p21ras gene products was evaluated immunohistochemically in ovarian carcinomas, borderline, and benign neoplasms. All studies were performed on cytospin preparations of cyst and/or ascitic fluid cells and mutual relations between oncoproteins were analysed. p53 and c-erbB-2 immunostaining was observed in 50% and 48.5% of ovarian carcinomas and in 30% and 35% of ovarian borderline tumors respectively, however in the last group the intensity and percentage of stained cells were considerably lower. In ovarian benign neoplasms there was no evidence of p53 and/or c-erbB-2 expression. The trend for serous carcinoma to have a higher p53 and c-erbB-2 expression than endometrioid and mucinous carcinomas was observed. p53 and c-erbB-2 oncoproteins were detected more frequently in the III/IV than in the I/II stages of the disease. The expression of p21ras was detected in 91% of malignant, 65% of borderline and 50% of benign neoplasms. p21ras reactivity was independent of the histopathological structure of ovarian carcinomas and it was comparable in I/II and III/IV FIGO stages. Our results indicate that p21ras overexpression appears to be an early genetic alteration in ovarian tumorigenesis, followed by the appearance of p53 and c-erbB-2 oncoproteins. It is likely that enhanced p53 and c-erbB-2 expression may cooperate with ras gene activation to produce a particularly aggressive phenotype. Our study supports the concept that development of ovarian carcinoma is the end result of a complex multistep process involving the complementary action of different cancer causing genes.

nm23 expression in tissue sections and tumor effusion cells of ovarian neoplasms.

The genetic changes involved in the metastatic process of ovarian epithelial cancer remain undetermined. The expression of nm23, a putative metastasis-suppressor gene product, was assessed immunohistochemically in malignant and benign ovarian neoplasms, considering histology of tumors and clinical advancement of disease. Comparison of nm23 protein content in tissue sections and respective cyst and/or ascitic fluid cells was also performed. Significant heterogeneity of nm23 immunostaining was observed, and no correlation with histological subtype of ovarian carcinoma was found. Expression of nm23 was higher in carcinomas compared with benign tumors. A significant trend to have a higher nm23 reactivity in ascitic fluid cells vs. primary tumors was observed. Our results indicate that the increase of nm23 reactivity is activated in the early stages of the disease and that the progression of ovarian carcinoma is accompanied by overexpression of nm23 protein. Our observations did not confirm the postulated role of nm23 as a suppressor gene in ovarian cancer.

[Studies of human papillomavirus infection (HPV) and expression of oncoprotein p 21 in cervix neoplasms]. / Badania nad zakazeniem wirusem ludzkiego brodawczaka (HPV) i ekspresja onkoproteiny p 21 w rakach szyjki macicy.

Frequencies of HPV type 16 and 18, evaluated by in situ hybridization technique in uterine cervix carcinomas of the IIIrd stage of clinical advancement, were 54% and 36.5% respectively. Presence of p 21 protein was detected in 85.3% cases of the cancers and showed no relation to HPV infection.

Relationship between c-erbB-2 oncoprotein, epidermal growth factor receptor, and estrogen receptor expression in patients with ductal breast carcinoma. Association with tumor phenotypes.

The expression of c-erbB-2 oncoprotein, epidermal growth factor receptor (EGFR) and estrogen receptor (ER) was evaluated by the immunoperoxidase technique (PAP) in ductal breast carcinomas. The relationship between these cell growth regulatory factors was considered and compared with tumor grading, tumor size, lymph node involvement and age of patients. Stratifying of patients on the basis of c-erbB-2, EGFR and ER status indicated that the combination of c-erbB-2 overexpression accompanied by high EGFR value and undetectable ER, identified poorly differentiated tumors and patients with high incidence of axillary lymph node metastases, while high EGFR expression and negative c-erbB-2 staining was connected only with poor tumor grade. The undetectability of molecular markers was associated with higher histological grade and lack of lymph node involvement. Our results indicate that the comparison of c-erbB-2, EGFR and ER status seems to be a powerful tool in discriminating breast carcinomas with different biological phenotypes.

Expression of p53 protein and Ki-67 reactivity in ovarian neoplasms. Correlation with histopathology.

The expression of Ki-67 proliferation antigen and its relation to p53 protein was assessed immunohistochemically in malignant and benign ovarian neoplasms, considering the stage of disease and histology of tumors. The comparison of p53 and Ki-67 in tissue sections and respective cyst and/or ascitic fluid cells was also performed. Significant heterogeneity of staining was observed. However, the relationship between p53 and Ki-67 activity in tissue sections and loose cells in individual patients was evident. Moreover, the presence of Ki-67 antigen was closely correlated with p53 protein. It was observed the trend for serous carcinoma to have a higher Ki-67 and p53 positivity versus endometrioid and mucinous carcinomas. However, it was not statistically significant. Both growth fraction as measured by Ki-67 staining and p53 content were significantly higher in stages III and IV compared to stages I and II of ovarian carcinomas (P<.05, and P<.01, respectively). In benign ovarian neoplasms, no p53 reactivity was observed and Ki-67 staining was very low. Our results showed that p53 is not a feature of benign epithelial ovarian tumors and indicate that increased proliferative activity of cells seems to involve immunohistochemically detectable alterations in p53 gene contributing to the evolution of ovarian carcinoma.

Relation between infection with human papilloma viruses and expression of p21 and p53 oncoproteins in patients with cancer of the uterine cervix.

Infection with HPV type 16 was demonstrated in 54% of the cases and infection with HPV type 18 in 36.5% of the cases of uterine cervix carcinoma. Both types of viruses were present in 24.3% of the patients. P53 protein accumulation in cell nuclei was observed in 12.2% of the cases of uterine cervix carcinomas and the cases were mostly HPV negative while the cytoplasmic expression of p53 protein was present in 39% of the cases and was frequently accompanied by HPV infection. Presence of p21 protein was detected in 85.3% of the cases, independently of HPV infection or expression of p53 protein.

Studies on tumor proliferation using monoclonal antibody, Ki 67 and expression of p53 in cancer of the uterine cervix.

The analyzed material included 41 sections originating from patients with squamous cell carcinoma at the III stage of clinical advancement. Size of tumor growth fractions and expression of p53 were evaluated using immunohistochemical methods. In most of cases, the Ki 67 index for uterine cervix carcinoma at the III stage of clinical advancement ranged between 5 and 30%. The p53 protein could seldom be detected, indicating infrequent p53 mutations in uterine cervix carcinoma. In cases exhibiting nuclear accumulation of p53 the proliferative index was high and ranged between 50% and 80%.

Cytomorphologic characterization of cell subsets isolated by density gradient centrifugation from tumor effusions of ovarian endometrioid carcinoma.

AIMS: Cytomorphologic characterization of tumor cell subsets, according to the stage of pathologic differentiation, and comparison of cellular composition in tumor cyst and ascitic fluids were carried out on individual patients with ovarian endometrioid carcinoma. METHODS: A density gradient centrifugation technique was applied to fractionate the cells from tumor effusions. RESULTS: The enrichment of cell forms representing individual stages of pathologic differentiation by gradient centrifugation facilitated their cytomorphologic characterization. According to cytomorphologic features, 5 discrete cell subpopulations were identified and catalogued. The cellular composition of tumor cyst and ascitic fluids in individual patients was similar, but the number of fractions and percentage of cell subsets differed. CONCLUSIONS: The estimation of precise cytomorphologic criteria for cell forms in tumor effusions facilitated the cytologic diagnosis of ovarian endometrioid carcinoma. The possibility to concentrate poorly differentiated, frankly malignant cell subsets in low densities could significantly improve the diagnosis of tumor effusions.

Relation between ovarian carcinoma-associated antigens in tumor tissue and detached cyst fluid cells of patients with ovarian neoplasms.

AIMS: The expression and potential diagnostic value of ovarian carcinoma-associated antigens were estimated in different types of epithelial ovarian neoplasms. The comparison of antigenic expression was performed on solid tumor tissues and loose cyst fluid cells in individual cases of malignant and benign ovarian neoplasms. METHODS: All studies were performed using monoclonal antibodies (mAbs) against ovarian carcinoma-associated antigens (OC125, OV-TL3, OV632, 10B, 8C) by 3-step peroxidase-antiperoxidase test. RESULTS: All ovarian carcinoma-associated antigens were detected in most serous and endometrioid carcinomas. In mucinous carcinomas as well as in benign ovarian neoplasms these antigens were present only in some cases. Significant inter- and intratumoral immunological heterogeneity was evident; however, the antigens detectable in tissue sections were also found in detached cyst fluid cells. CONCLUSIONS: Our results show that mAb show OV-TL3 is the best marker for endometrioid carcinomas and confirmed that mAbs OV632, OC125 and OV-TL3 could be good complementary markers for differentiating malignant and benign lesions in the ovary. The percentage content of all ovarian carcinoma-associated antigens in solid tumors and respective cyst fluid cells was comparable.

Relationship between p53 and c-erbB-2 overexpression in tissue sections and cyst fluid cells of patients with ovarian cancer.

The expression of p53 protein and overexpression of c-erbB-2 oncoprotein was examined immunohistochemically and compared on frozen tissue sections and cyst fluid cells in patients with epithelial malignant and benign ovarian neoplasms. p53 was detected in 52.6% of carcinomas and c-erbB-2 expression was identified in 47.3% of cases. The relationship between p53 and c-erbB-2 overexpression in tissue sections and detached cyst fluid cells was evident. Moreover, a significant association between the presence of p53 and overexpression of c-erbB-2 proteins in tumor tissue sections and loose cyst fluid cells in individual patients was demonstrated. In tissue sections and loose cyst fluid cells of benign ovarian tumors no overexpression of p53 and c-erbB-2 proteins was found.

Carcinoembryonic antigen isotypes in tissue sections and loose cyst fluid cells of ovarian neoplasms.

The aim of this study was to establish whether different subsets of ovarian neoplasms express a restricted isotype of carcinoembryonic antigen (CEA) which can be detected in solid tumors and detached cells. Sixty-one cases of mucinous, serous, endometrioid, and Krukenberg tumors were studied by immunohistochemistry using two monoclonal antibodies (MAbs), commercial anti-CEA and D14 with a higher specificity for colorectal adenocarcinomas. The results with both antibodies showed a considerable degree of heterogeneity between cases of nonserous tumors, with a more restrictive pattern observed with the D14 MAb. The proportion of immunostained cells was comparable in tumors and fluids.

Comparison of cellular phenotypes in tumor cyst and ascitic fluid from ovarian serous carcinoma.

Density gradient centrifugation was applied to isolate cell subsets from tumor cyst and ascitic fluid in eight patients with ovarian serous carcinoma. A comparison of cellular composition and immunologic reactivity of cells from the cysts and from ascitic fluid in each patient was performed. Some differences in density profiles were found, but in each case the consistency of morphologic cell forms in the primary tumor and ascites was documented. Immunophenotypic analyses of isolated cellular fractions using polyclonal and monoclonal antibodies against ovarian carcinoma-associated antigens showed significant immunologic intratumoral heterogeneity. However, there was a similarity of antigen expression in cells from the primary tumors and ascitic fluids. Our study indicated that morphologic and antigenic characterization of a given tumor could be determined in a single representative sample of ascitic fluid.

Density distribution and immunological reactivity of tumor cells from peritoneal effusions of patients with ovarian neoplasms.

Ascitic samples from 19 patients with primary ovarian non-mucinous carcinomas, three with Krukenberg tumors and eight with noncancerous peritoneal effusions were studied by conventional cytology and immunocytochemical staining. Density gradient centrifugation was applied to fractionate ascitic fluid cells. The enrichment of cell types by this method facilitated their cytomorphological characterization and identification of neoplastic cell subpopulations existing in peritoneal effusions. Immunophenotypic studies of cells were made using monoclonal antibodies (mAbs) against ovarian carcinoma-associated antigens (OC 125, 10B, 8C) and carcino-embryonic antigen (CEA). Non-specific cross-reacting antigen (NCA) was applied as a marker for granulocytes which often accompany peritoneal effusions. Our results indicated that immunofluorescence (IF) staining contributed to the distinction between the primary and secondary ovarian carcinomas. Density gradient centrifugation appeared to be a useful method for separation of mesothelial cells.