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.

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.

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.

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.

Morphological and immunological heterogeneity of human ovarian neoplasms.

The cells of cystic fluids from patients with malignant and benign serous ovarian neoplasms were separated by density gradient centrifugation. Density distribution and morphological features of cell fractions were analyzed. The immunophenotypic characterization of each cell fraction using poly- and monoclonal antibodies was performed. The application of density gradient centrifugation allowed to yield and catalogue several cell subpopulations according to their stage of pathological differentiation starting from typical, morphologically malignant until normal epithelial cells. Significant morphological and immunological heterogeneity among and within individual tumors was found. Our studies indicated the value of isolated cell subsets for estimation of interrelationships among various neoplastic markers and for comparison the reactivity of different antibodies and evaluation of their immunodiagnostic potency.

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.

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.

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.

Density distribution and antigenic characterization of cell subpopulations isolated from cystic fluids of ovarian serous neoplasms.

The immunological reactivity of serous ovarian tumor cells was evaluated, taking into account their density and morphological features. Discontinuous density gradient centrifugation was applied to fractionate cell subpopulations from cystic fluids of patients with ovarian serous carcinomas, cystadenomas and benign serous cysts. For phenotypic characterization of tumor cell subpopulations the immune sera against perchloric acid extracts of ovarian serous (anti-PCA-CaOs) and mucinous (anti-PCA-CaOm) carcinomas were used. The expression of carcinoembryonic antigen (CEA) and nonspecific cross-reacting antigen (NCA) on tumor cell fractions was also checked. Some relationship between immunological reactivity, cell morphology and cell density was found; however, individual patient-to-patient variations in cellular composition and antigenic expression were also observed. The presence of ovarian serous carcinoma-associated antigen (CaOs-Ag) was related mainly to frankly malignant cells. Neither CEA nor NCA were constant and characteristic markers for serous ovarian neoplasms. These neoplasms were unreactive with anti-PCA-CaOm serum. Our results indicate the possibility of correlation between morphological features, density distribution and immunological phenotypes of ovarian tumor cell populations.

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.

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.

Morphologic heterogeneity of cell populations isolated by density gradient centrifugation from serous fluids of ovarian tumors.

The cells of tumor fluid from patients with malignant and benign serous ovarian neoplasms were fractionated using Ficoll-Uropoline density gradient centrifugation. Density distribution and morphologic characteristics of cell fractions were analyzed. It was found that serous ovarian adenocarcinomas contained three to four types of morphologically malignant cells focused in low density layers. Borderline ovarian neoplasms showed the presence of one subpopulation of cells with some features of malignancy and cells with some atypical but non-malignant features. The fluids of serous cysts contained mainly normal epithelial cells representing different stages of morphological maturity and were focused in denser layers. The results allowed us to catalogue ovarian tumor cell subpopulations present in each density fraction of individual patients and confirmed that ovarian tumors could be diagnosed by morphologic identification of cells from tumor fluids.

Reactivity of polyclonal and two monoclonal antibodies with cell subsets isolated from cystic fluids of ovarian serous neoplasms.

The distribution of antigenic determinants on a cellular level in serous ovarian neoplasms was evaluated using polyclonal and two monoclonal antibodies (OC 125 and 10B). The expression of antigens was estimated by an immunofluorescence test on each cell fraction isolated by density centrifugation from cystic fluids of individual malignant and benign ovarian tumors, taking into account the density and cytomorphologic features of cell subpopulations. It was found that the studied antibodies recognized different antigenic determinants. Significant immunologic heterogeneity of cells among and within individual tumors was shown. Our studies show the value of isolated cell subpopulations for comparing the reactivity of different antibodies and estimating their immunodiagnostic potency.

Density distribution, cytomorphologic features and immunologic characteristics of ovarian and endometrial clear cell carcinomas.

Three subpopulations of cancer cells with different morphologic features were separated by density gradient centrifugation of two ascitic fluids and one cystic fluid from one patient with ovarian clear cell carcinoma and one with endometrial clear cell carcinoma. Immunophenotypic analyses of isolated fractions using polyclonal and monoclonal antibodies against ovarian carcinoma-associated antigens revealed significant immunologic heterogeneity among the tumor cells. The identical histopathologic structures of the ovarian and endometrial clear cell carcinomas and the similar distribution and immunologic reactivity of the cell types isolated from the ascitic and cystic fluids confirmed the common histogenesis of both cancers. These findings suggest that the conventional cytologic diagnosis of clear cell carcinomas could be supplemented by immunofluorescent staining. Density gradient centrifugation appeared to be a useful method for the separation of mesothelial cells.

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.

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%.

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 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 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.