Miriam Aparecida da Silva Trevisan

The effect upon the human vaginal histology of the long-term use of the injectable contraceptive Depo-Provera®☆

The objective of the study was to evaluate the effect of long-term use of the injectable contraceptive depot medroxyprogesterone acetate (DMPA) on human vaginal histology. Twenty premenopausal women currently using DMPA as a contraceptive method for two and three years were compared with 20 regularly menstruating women, who never used Depo-Provera and/or other kind of hormonal contraceptive in the last 6 months prior to the study. Subjects and controls were matched by age (+/-1 year), body mass index (kg/m2) (+/-1.0), number of pregnancies (+/-1), age at first intercourse (+/-1 year), years of sexual activity (+/-1 year), and number of partners during their life (+/-1). Vaginal biopsies were performed in users at 90+/-7 days after the last injection and in nonusers at day 20-25 of the menstrual cycle. In addition, at the day of the biopsy a blood sample was collected to measure estradiol (in all women) and DMPA in users. The level of serum estradiol was significant lower in Depo-Provera users than in controls (p < 0.001). The thickness of the vaginal epithelium was not smaller among DMPA users than among controls, the mean count of Langerhans cells per mm of epithelium were almost identical in both groups, and no significant differences were found on the vaginal maturation indices. In conclusion, the use of Depo-Provera between two and three years did not affect vaginal thinning of the epithelium, Langerhans cell count or maturation index.

THE VALUE OF PCNA AND AGNOR STAINING IN ENDOSCOPIC BIOPSIES OF GASTRIC MUCOSA

The aim of the present study was to examine the usefulness of the quantification of PC10-positive-cells and of Argyrophilic Nucleolar Organizer Regions (AgNORs) in gastric biopsies for the identification of gastric mucosal proliferative lesions. Fifty seven paraffin-embedded endoscopic biopsies were classified into four histologic groups: normal, inflammatory, dysplastic and neoplastic mucosa. The percentage of PC10-positive cells was determined by immunohistochemistry. The AgNOR parameters determined included the total number of all identifiable silver precipitations in the nucleus, the mean number of silver precipitations per cluster, and the presence of morphologically heterogenous silver precipitations. Group comparisons were performed using the Kruskall Wallis and Dunn non-parametric tests with a significance level of 5%. A discriminant analysis (followed by the jack-knife procedure) was performed using the three AgNOR parameters plus the percentage of PCNA-positive cells as the independent variables and histological groups as the dependent variable. All three AgNOR parameters, as well as the percentage of PCNA-stained nuclei, showed their highest values in the carcinoma group. However, no good differentiation among the four histologic groups was obtained using only one of these parameters, since there was always considerable overlap among them. By combining all the parameters in a linear discriminant analysis, we obtained a correct classification in 48 out of 57 cases. Within the classification errors there was only one false positive carcinoma, which was in fact a dysplasia and only one false negative carcinoma erroneously classified as dysplasia. The number of cells with heterogenous AgNORs was the most important parameter for the discriminant analysis. No correlation between PCNA values and the AgNOR parameters could be found, thus indicating that they do not represent the same phenomenon in the cell cycle. We concluded that the use of a combination of various proliferation parameters in a linear discriminant analysis may be helpful for differentiating gastric mucosal lesions. The peculiar AgNOR morphology is an important variable which should be taken in consideration in quantitative studies. PCNA and AgNORs seem to represent different physiological phenomena in the cell cycle.

Immunohistochemical analysis of vascular density and area in colorectal carcinoma using different markers and comparison with clinicopathologic prognostic factors

Analysis of blood and lymphatic vessel in colorectal cancer is controversial in the literature, possibly due to variations in the methods of analysis. In this study, it was aimed to search for a reliable approach in the quantification of angio- and lymphangiovascular density and area as a prognostic factor and to compare such vessel counts in normal mucosa, adenomas and cancer. A retrospective study was performed on 60 sporadic colorectal cancer, 30 colorectal adenomas, and 10 colorectal non-neoplastic lesions. Archival tissues were submitted to immunohistochemical evaluation using antibodies to CD31, CD34, CD105, VEGF-A, VEGF-C, and D2-40. Microvessel density and total vascular area were determined by computer image analysis and values were compared in the three groups of lesions; the prognostic value of these parameters was evaluated in the group of colorectal cancer. Most markers showed progressive vessel counts from non-neoplastic tissue to carcinoma, both for microvessel density and total vascular area. Only microvessel density determined by CD34 in the central areas of the cancer correlated with recurrence/metastasis (p = 0.04) and survival (p = 0.02). Different methods of quantification (microvessel counting versus estimation of total vascular area), immunohistochemical markers (pan-endothelial marker versus neovessels and lymphatic markers), and areas of analysis (periphery versus inner portions of the lesion) were assessed using image analysis. The results corroborate the increase in vascularization of carcinoma and suggest that microvessel density determined by immunostaining for CD34 in the inner portion of the tumor might represent a prognostically relevant parameter in colorectal cancer.

CagA status and Helicobacter pylori eradication among dyspeptic patients.

AIM Triple therapy seems more effective in curing Helicobacter pylori infection in patients with peptic ulcer than in those with non-ulcer dyspepsia. It has been suggested that this difference depends on the expression of CagA protein that is more frequent in the former. The objective of this study was to investigate a potential association between serum CagA positivity, severity of gastric mucosal inflammation and eradication success among peptic ulcer and non-ulcer dyspepsia patients. MATERIAL AND METHOD Patients undergoing upper gastrointestinal endoscopy for investigation of dyspepsia at the Department of Gastroenterology, Hospital Vera Cruz, between March, 2000 and March 2001 were screened. H. pylori positive patients, as diagnosed by rapid urease test and histology were included. Severity of gastric mucosal inflammation was determined and serum CagA positivity was assessed using a commercially available ELISA assay prior to H. pylori 7-day eradication therapy with lansoprazole, clarithromycin and amoxicillin (30 mg, 500 mg and 1 g b.i.d., respectively). Eradication success was determined 8-24 weeks following completion of therapy. RESULTS Seventy-four patients were included in the study (mean age 40.8, range 18-67, female = 28). CagA positivity was observed in 48% of patients. Gastroduodenal peptic ulceration was found in 54% of patients. Serum CagA positivity was significantly higher among peptic ulcer patients (62.5%), while CagA negativity was significantly higher among non-ulcer dyspepsia patients (67.7%). Lymphocyte and eosinophil infiltration was significantly higher among CagA + patients, despite being comparable when distributed among peptic ulcer and non-ulcer dyspepsia patients. Eradication was successful in 93.2% of patients, regardless of CagA status on a per protocol analysis. Based on a per protocol analysis, eradication success was comparable among peptic ulcer and non-ulcer dyspepsia patients, regardless of CagA status. CONCLUSION Our results support the concept that CagA positivity is associated to peptic ulcer disease and to a higher severity of lymphocyte and eosinophil infiltration. Efficacy of treatment eradication of H. pylori may not be affected by serum CagA status.

Menstrual pattern and ovarian function in women with hyperprolactinemia

Prolactine (PRL) and progesterone were systematically measured in all women presenting with amenorrhea, oligomenorrhea or galactorrhea at the Infertility Clinic of the Department of Obstetrics and Gynecology, State University of Campinas, Brazil, during a period of 34 months. The same hormonal assays were done to all infertile patients presenting for the first time during the last 6 months of the same period, for a total of 190 subjects. Fifty‐five patients with amenorrhea, 38 with oligomenorrhea and 97 with normal cycles, 20 of whom had galactorrhea, were included in the study. Fifty‐five percent of amenorrheic patients, 37% of oligomenorrheic and 9% of those with normal menses had elevated PRL. The mean PRL was higher the greater the menstrual disturbance but was not influenced by presence or absence of galactorrhea. Short luteal phase was the ovarian function condition most frequently associated with high PRL among women with normal menses.

Correlation Between Thymidylate Synthase Protein Expression and Gene Polymorphism With Clinicopathological Parameters in Colorectal Carcinoma

5-Fluorouracil (5-FU) represents the basis of chemotherapy for colorectal carcinoma, inhibiting thymidylate synthase (TS), an essential enzyme for DNA replication. Previous studies have associated high TS protein expression by tumor cells with poor outcome of patients with colorectal carcinoma, but others have refuted these findings. In view of the potential role of TS as predictive parameter and the lack of consensus in the literature, the present study compared 2 methods: protein expression and gene polymorphism, correlating them with clinicopathological findings. Immunohistochemical detection of TS in tumor cells and detection of gene polymorphism in the blood were performed in 32 patients with colorectal carcinoma treated with 5-FU. No correlation was found between TS protein expression and gene polymorphism. Neither method correlated with survival, tumor staging, and tumor histological grading. This result possibly reflects a complex tumor response to 5-FU therapy, where TS is just one of the involved proteins.

Necrotic Changes of the Adrenal Cortex Following Orthostatic Collapse in Rabbits

The adreal gland was studied in 56 adult male rabbits; 44 had been subjected to orthostatic collapse, and 12 served as controls. The animals either died from collapse or were sacrificed at regular intervals up to 30 days after the experimental procedure. Nuclear pyknosis was seen in cortical cells of animals dying in collapse. At 6 or more hours following collapse, necrosis was found involving single cells, or rarely small groups of cells mainly in the zona fasciculata. Regenerative areas were present at 7 and 15 days after collapse. This necrosis may be related to a nutritive disturbance, largely hypoxic in nature (hypovolemia with blood stasis and thrombosis). The adrenal medulla showed only degranulation.

Necrosis of gastric mucosa following orthostatic collapse in rabbits

Twenty-five rabbits were submitted to orthostatic collapse, divided into 6 groups and sacrificed about 4, 24 and 48 hours, 8, 15 and 30 days following collapse; two animals died during collapse and another 72 hours later. Seven animals were used as controls. Necrotic changes involving small areas of the gastric mucosa in the fundic and body portions were seen in successive stages up to regeneration. Such changes were present in all animals submitted to collapse, except three with no lesion. The necrosis is related to a nutritive disturbance of the gastric mucosa, mainly anoxic in nature. Grossly, the necrotic areas were similar to the acute erosions of the gastric mucosa seen in man.

Comparison of blood neoangiogenesis and lymphatic vascularization in colorectal adenomas from patients with and without concomitant colorectal cancer

Blood and lymphatic vessel proliferation is essential for tumor growth and progression. Most colorectal carcinomas develop from adenomas (adenoma-carcinoma sequence) in a process due to accumulation of molecular genetic alterations. About 5% of adenomatous polyps are expected to become malignant, but data on the differential angiogenic patterns of these lesions in patients with and without concomitant cancer are missing. The aim of the present study is to compare the angiogenic and lymphatic patterns of adenomatous polyps from patients with and without sporadic cancer. Thirty adenomatous polyps (15 from patients with another principal malignant lesion, and 15 from patients without cancer) were submitted to immunohistochemical staining for CD105 (marker for neoangiogenesis) and D2-40 (marker for lymphatic endothelium). Microvessel density and total vascular area were determined by computer image analysis to quantify the immunostained and total areas, and to assess the number of microvessels. Adenomas from patients with carcinoma showed significantly higher values of total vascular area determined by immunostaining for CD105 (cutoff value = 4386 microm(2); P = 0.019) and of lymphatic microvessel density determined by immunostaining with D2-40 (cutoff value = 11.5; P = 0.041) when compared with those from patients without cancer. The present data indicate a significant increase in blood microvascular area and in lymphatic microvascular counts in adenomas removed from patients with cancer.

A inter-relação das sialomucinas (antígenos Tn e Stn) com o adenocarcinoma no esôfago de Barrett

OBJECIVE: Barretts esophagus (BE) is a consequence of chronic gastroesophageal reflux and is considered a risk factor for adenocarcinoma. The study of the mucus, especially acid mucins, such as the sialomucins in the goblet cells which characterize BE, showed that in intestinal metaplasia, frequent in the digestive tract, the organs original epithelium could express Tn and Stn antigens. These antigens have already been detected in gastric and colonic tumors, however references in BE were not found. This research aimed to analyze these antigens in patients with BE and in adenocarcinoma associated with BE. METHODS: Utilizing immunohistochemistry tests, Tn and Stn antigens were studied in the endoscopic biopsies of 29 patients with BE and seven with adenocarcinoma in BE, as well as eight individuals with normal esophageal epithelium at upper digestive endoscopy.. RESULTS: The Stn antigen was positive in the goblet cells of patients with BE in 100% of the cases and the Tn was positive in 48%. In the columnar cells, Stn was always negative, while Tn was positive in 100% of the cases. However, in adenocarcinoma in BE, both antigens were 100% positive. In normal individuals, the Tn antigen was positive and the antigen Stn negative in all cases. CONCLUSION: It is probable that the BE group in which the Tn antigens in the goblet cells are positive, similarly to the same antigen in the adenocarcinoma group, might indicate a higher susceptibility for potential occurrence of cancer. In the future, trials with sialomucins could be used routinely, thereby contributing as a prognostic factor of adenocarcinoma in BE.