Grape Juice Concentrate Protects Rat Liver Against Cadmium Intoxication: Histopathology, Cytochrome C and Metalloproteinases Expression.

The aim of this study was to investigate if grape juice concentrate is able to protect rat liver against cadmium toxicity. For this purpose, histopathological analysis, cytochrome C expression and immunoexpresssion of metalloproteinases (MMP) 2 and 9 were investigated. A total of 15 Wistar rats weighing 250 g on the average, and 8 weeks age were distributed into 3 groups (n=5), as follows: Control group (non-treated group, CTRL); Cadmium group (Cd) and grape juice concentrate group (Cd+GJ). Histopathological analysis revealed that liver from animals treated with grape juice concentrate improved tissue degeneration induced by cadmium intoxication. Animals intoxicated with cadmium and treated with grape juice concentrate showed higher cytochrome C gene expression in liver cells. No significant statistically differences (p>0.05) were found to MMP 2 and 9 immunoexpression between groups. Taken together, our results demonstrate that grape juice concentrate is able to prevent tissue degeneration in rat liver as a result of increasing apoptosis.

Application of fluorescence in situ hybridization (FISH) as a tool to aid cytogenetics in 1,409 fetal samples.

AIM: To evaluate the technical application of fluorescence in situ hybridization (FISH) as a support to classical cytogenetic in numerical chromosomal aneuploidies studies in samples of amniotic fluid, chorionic villus, and fetal loss. MATERIALS AND METHODS: The authois performed cytogenetic analyses in 1,409 patients (678 amniocentesis, 512 chorionic villus samples, and 219 spontaneous abortions) during one year. FISH molecular study aided traditional cytogenetic in 90 cases. These cases were indicated based on the diagnostic hypothesis of each patient or when no cellular growth was obtained. The authors standardized the FISH in discoloured slides. RESULTS: They had 85% positive FISH in amniotic fluid, 70% in chorionic villus, and 90% in abortion material using 13, 18, 21 X and Y centromeric probes. It showed 12% of altered FISH in amniotic fluid (100% trisomies), 10% in chorionic'villus (50% trisomy and 50% X - monosomy), and 22% in abortion material (50% trisomy, 25% X-monosomy, and 25% triploidy). FISH and cytogenetic analysis confirmed the results. CONCLUSION: This technique revolutionized clinical and research applications of cytogenetics. In this particular paper, FISH was a valuable and reliable technique to promptly identify rapid detection of aneuploidies in interphase cells, metaphase spread and paraffin-embedded samples. It is hoped that, in the future, the economic viability of array CGH and FISH, with the decreasing cost of testing and their genomics advantages can be incorporated as routine and customized in the approach of prenatal diagnosis.

Expression of apoptosis regulatory proteins p53, bcl-2 and bax in recurrent aphthous ulceration.

BACKGROUND: Recurrent aphthous ulceration (RAU) is considered to be an acute inflammatory disease of unknown pathogenesis. Apoptosis may represent an important event in the control of inflammation. OBJECTIVES: The aim of this study was to investigate apoptosis process in RAU using immunohistochemistry. METHODS: We studied the expression and location of p53, bcl-2 and bax in ulcerated lesions clinically diagnosed as RAU (n = 12) and compared it with that of oral clinically normal mucosa (n = 6) and of other inflammatory chronic disease such as oral fibrous inflammatory hyperplasia (FIH; n = 18). RESULTS: Significant statistically differences (n < 0.05) in p53 expression were noticed in RAU when compared with normal mucosa. No significant statistically differences (P > 0.05) were noticed between FIH and RAU. Bcl-2 and bax did not show remarkable differences between groups. CONCLUSIONS: Taken together, the data suggest that RAU induces p53 immunoexpression. Therefore, the protein might be related to the aetiopathogenesis of the ulcerated oral lesions.

Morphocytochemical, immunohistochemical and ultrastructural characterization of the head kidney of fat snook Centropomus parallelus.

This study characterized the structure and the morphocytochemical, immunohistochemical and ultrastructural aspects of the head kidney (HK) of the fat snook Centropomus parallelus. The HK is enclosed by a thin capsule of connective tissue, from which fine trabeculae originate and branch into the interior of organ. In the parenchyma, there are aggregates of lymphoid cells containing populations of lymphocytes T immunopositive for CDRO45, in a nodular arrangement, around blood vessels and melano-macrophage centres. Among the cells that constituted these aggregates and surrounded them, were macrophages and monocytes, and their precursors, with strong immunopositivity for CD68, along with cells of the granulocytic lineage in various phases of maturation positive for lysozyme and PAS. Macrophages and chromaffin and interrenal cells are also present. Ultrastructurally, the HK comprises a reticulum-endothelial stroma consisting of endothelial cells, reticulocytes of the fibroblast type and macrophage type and a parenchyma with increased cellularity, principally blood cells of the erythrocytic, granulocytic, lymphocytic, monocytic and thrombocytic series.

Immunohistochemical expression of types I and III collagen antibodies in the temporomandibular joint disc of human foetuses.

The objective was to study the morphology of the articular disc and analyse the immunohistochemical expression of types I and III collagen markers in the temporomandibular joint (TMJ) disc of human foetuses of different gestational ages. Twenty TMJ from human foetuses supplied by Universidade Federal de Uberaba with gestational ages from 17 to 24 weeks were studied. The gestational age of the foetuses was determined by measuring the crown-rump (CR) length. Macroscopically, the foetuses were fixed in 10% formalin solution and dissected by removing the skin and subcutaneous tissue and exposing the deep structures. Immunohistochemical markers of type I and III were used to characterize the existence of collagen fibres. Analysis of the immunohistochemical markers of types I and III collagen revealed the presence of heterotypical fibril networks.

Biomonitoring of DNA damage and cytotoxicity in individuals exposed to cone beam computed tomography.

OBJECTIVES: The aim of the present study was to evaluate DNA damage (micronucleus) and cellular death (pyknosis, karyolysis and karyorrhexis) in exfoliated buccal mucosa cells from adults following cone beam CT exposure. METHODS: A total of 19 healthy adults (10 men and 9 women) submitted to cone beam CT were included. RESULTS: No significant statistically differences (P > 0.05) in micronucleus frequency were seen before and after cone beam CT exposure. In contrast, the tomography was able to increase other nuclear alterations closely related to cytotoxicity such as karyorrhexis, pyknosis and karyolysis (P < 0.05). CONCLUSION: In summary, these data indicate that cone beam CT may not be a factor that induces chromosomal damage, but it is able to promote cytotoxicity.

Over-expression of cyclooxygenase-2 in endoscopic biopsies of ectopic gastric mucosa

Ectopic gastric mucosa (EGM) is considered to be a congenital condition. Rare cases of adenocarcinoma have been described. There are no data justifying regular biopsies or follow-up. Cyclooxygenase-2 (COX-2) is a protein involved in gastrointestinal tumor development by inhibiting apoptosis and regulating angiogenesis. The aim of this prospective study was to evaluate COX-2 expression in EGM and compare it with normal tissue and Barrett's esophagus. We evaluated 1327 patients. Biopsies were taken from the inlet patch for histological evaluation and from the gastric antrum to assess Helicobacter pylori infection. Biopsies taken from normal esophageal, gastric antrum and body mucosa and Barrett's esophagus were retrieved from a tissue bank. EGM biopsies were evaluated with respect to type of epithelium, presence of H. pylori, and inflammation. COX-2 was detected by immunohistochemistry using the avidin-biotin complex. EGM islets were found in 14 patients (1.1 percent). Histological examination revealed fundic type epithelium in 58.3 percent of cases, H. pylori was present in 50 percent and chronic inflammation in 66.7 percent. Expression of COX-2 was negative in normal distal esophagus, normal gastric antrum and normal gastric body specimens (10 each). In contrast, EGM presented over-expression of COX-2 in 41.7 percent of cases and Barrett's esophagus in 90 percent of cases (P = 0.04 and 0.03, respectively). COX-2 immunoexpression in EGM was not related to gender, age, epithelium type, presence of inflammation or intestinal metaplasia, H. pylori infection, or any endoscopic finding. Our results demonstrate up-regulation of COX-2 in EGM, suggesting a possible malignant potential of this so-called harmless mucosa.

Over-expression of cyclooxygenase-2 in endoscopic biopsies of ectopic gastric mucosa.

Ectopic gastric mucosa (EGM) is considered to be a congenital condition. Rare cases of adenocarcinoma have been described. There are no data justifying regular biopsies or follow-up. Cyclooxygenase-2 (COX-2) is a protein involved in gastrointestinal tumor development by inhibiting apoptosis and regulating angiogenesis. The aim of this prospective study was to evaluate COX-2 expression in EGM and compare it with normal tissue and Barrett's esophagus. We evaluated 1327 patients. Biopsies were taken from the inlet patch for histological evaluation and from the gastric antrum to assess Helicobacter pylori infection. Biopsies taken from normal esophageal, gastric antrum and body mucosa and Barrett's esophagus were retrieved from a tissue bank. EGM biopsies were evaluated with respect to type of epithelium, presence of H. pylori, and inflammation. COX-2 was detected by immunohistochemistry using the avidin-biotin complex. EGM islets were found in 14 patients (1.1%). Histological examination revealed fundic type epithelium in 58.3% of cases, H. pylori was present in 50% and chronic inflammation in 66.7%. Expression of COX-2 was negative in normal distal esophagus, normal gastric antrum and normal gastric body specimens (10 each). In contrast, EGM presented over-expression of COX-2 in 41.7% of cases and Barrett's esophagus in 90% of cases (P = 0.04 and 0.03, respectively). COX-2 immunoexpression in EGM was not related to gender, age, epithelium type, presence of inflammation or intestinal metaplasia, H. pylori infection, or any endoscopic finding. Our results demonstrate up-regulation of COX-2 in EGM, suggesting a possible malignant potential of this so-called harmless mucosa.

Immunoexpression of cyclooxygenase-1 and -2 in ulcerative colitis.

Ulcerative colitis (UC) is a disease of the colon and rectum characterized by a nonspecific chronic inflammation mediated by the concerted response of cellular and humoral events. Prostaglandins are synthesized by cyclooxygenase (COX)-1 and -2 and exhibit both pro- and anti-inflammatory activity. To evaluate COX-1 and COX-2 immunoexpression in 42 cases of UC and to correlate it with clinicopathological parameters, COX-1 and COX-2 expression was investigated by the immunohistochemistry method. Only patients with all pertinent clinical and evolutive data as well as with adequate biopsy material were included in the study. Fifteen samples of colorectal adenocarcinoma and 14 of large bowel with no histological changes were used for positive and negative controls, respectively. UC patients showed COX-1 immunoreactivity in epithelial cells in 29% of the cases and in inflammatory cells in 43%. COX-2 positivity in epithelial and inflammatory cells was found in 69% of the samples. The comparison between UC and the control groups revealed that the UC group had significantly more positive cases for COX-1 and COX-2 in inflammatory cells. Immunohistochemistry allowed the identification of COX-1 and COX-2 expression in epithelial and inflammatory cells in UC biopsies. No significant difference between COX-1 and COX-2 immunoreactivity in epithelial and inflammatory cells was observed regarding the clinicopathological parameters. COX-2 presented low expression in normal colon and high expression in colorectal adenocarcinoma. COX-2 might play a role in the pathophysiologic processes of inflammatory bowel disease and the development of neoplasia. Treatment with selective COX-2 inhibitors might be an additional option for therapy.

Immunoexpression of cyclooxygenase-1 and -2 in ulcerative colitis

Ulcerative colitis (UC) is a disease of the colon and rectum characterized by a nonspecific chronic inflammation mediated by the concerted response of cellular and humoral events. Prostaglandins are synthesized by cyclooxygenase (COX)-1 and -2 and exhibit both pro- and anti-inflammatory activity. To evaluate COX-1 and COX-2 immunoexpression in 42 cases of UC and to correlate it with clinicopathological parameters, COX-1 and COX-2 expression was investigated by the immunohistochemistry method. Only patients with all pertinent clinical and evolutive data as well as with adequate biopsy material were included in the study. Fifteen samples of colorectal adenocarcinoma and 14 of large bowel with no histological changes were used for positive and negative controls, respectively. UC patients showed COX-1 immunoreactivity in epithelial cells in 29 percent of the cases and in inflammatory cells in 43 percent. COX-2 positivity in epithelial and inflammatory cells was found in 69 percent of the samples. The comparison between UC and the control groups revealed that the UC group had significantly more positive cases for COX-1 and COX-2 in inflammatory cells. Immunohistochemistry allowed the identification of COX-1 and COX-2 expression in epithelial and inflammatory cells in UC biopsies. No significant difference between COX-1 and COX-2 immunoreactivity in epithelial and inflammatory cells was observed regarding the clinicopathological parameters. COX-2 presented low expression in normal colon and high expression in colorectal adenocarcinoma. COX-2 might play a role in the pathophysiologic processes of inflammatory bowel disease and the development of neoplasia. Treatment with selective COX-2 inhibitors might be an additional option for therapy.

Ki-67 as a prognostic marker in colorectal cancer but not in gastric cancer.

The growth of tumors is highly variable and this probably reflects even its clinical course. The monoclonal antibody Ki-67 recognises an antigen present in the nuclei of cells in all phases of the cell cycle except G0. In the current study, we examined by immunohistochemistry the proliferative activity, based on Ki-67 labeling index (Ki67LI), in formalin-fixed and paraffin-embedded sections of 152 tumors, being 70 gastric and 89 colorectal cancers. The results obtained were correlated with the clinicopathologic factors. The carcinomas showed a wide range of Ki-67LI, reflecting a variation in proliferative activity. The tumor labeling index ranged from 10 to 85 per cent positivity, being the mean level in gastric cancer tissue 0.52 and in colorectal cancer 0.44. There was also heterogeneity of labeling within many of the tumors. No significant correlation was found between Ki-67LI and sex, age, clinical stage in these cancers. In colorectal cancer, but not in gastric cancer, high levels of Ki67LI have been correlated with poor survival. Ki-67 staining is a simple and useful method for estimating proliferative activity. The importance of Ki-67 as an indicator of tumor behaviour is not clear. In colorectal cancer this index may be used as a marker of prognosis.

Early nuclear alterations and immunohistochemical expression of Ki-67, Erb-B2, vascular endothelial growth factor (VEGF), transforming growth factor (TGF-beta1) and integrine-linked kinase (ILK) two days after tamoxifen in breast carcinoma.

The purpose of the present study was to evaluate breast carcinoma samples before and two days after treatment with tamoxifen in order to analyse early histopathological alterations--particularlynuclear alterations-- as well as immunohistochemical expression of Ki-67, Erb-B2, VEGF, TGF-beta1 and ILK proteins. Twenty one cases of invasive ductal and lobular breast carcinoma were studied. Patients were submitted to biopsy of the lesion and, after confirmation of the diagnosis, they received 20 mg of tamoxifen a day, beginning two days before surgery. The samples obtained during biopsy and after surgery were stained with HE for histopathological diagnosis. Estrogen receptor was positive in 18 cases and negative in 3. The immunohistochemical method was applied for the detection of Ki-67, Erb-B2, protein, vascular endothelial growth factor (VEGF), transforming growth factor beta (TGF-beta1) and integrin linked kinase (ILK). Two days after tamoxifen treatment, the following results were observed: 1) decrease in the cell volume, chomatine condensation, nucleoli less evident and clearly defined nuclear limits; 2) significant reduction in the expression of Erb-B2 protein and significant increase in the expression of TGF-beta1 protein; 3) expression of others proteins (Ki-67, VEGF and ILK) was not altered during the indicated time frame. Our results suggest that analyzing nuclear alterations and expression of Erb-B2 and TGF-beta1 proteins would be useful to assess the initial response to tamoxifen.

Pinealectomy changes rat ovarian interstitial cell morphology and decreases progesterone receptor expression.

The aim of this study was to evaluate the rat ovarian morphological and function changes after pinealectomy (px). Two months after px, young female Wistar rats were sacrificed and the right ovaries were analysed morphologically and the left ovaries were used for steroid receptor binding experiments. Blood was collected and steroid hormone and melatonin levels were measured using radioimmunoassay kits. Results revealed that in the px group the rat ovaries had an increase in the number of atretic follicles and interstitial cells. These cells showed hyperactivity features on transmission electron microscopy and morphometric analysis (p < 0.05 compared with control and sham groups). Px-group serum showed an increase in estradiol (p < 0.05) and a decrease in progesterone levels (p < 0.05) compared with other groups. Moreover, progesterone receptor expression was lower than control and sham groups (p < 0.05). We postulate that pinealectomy leads to many morphological alterations of rat ovaries that are associated with functional changes in steroidogenesis and a decrease in progesterone receptor expression.

[AgNOR in stomach neoplasm]. / AgNOR em câncer gástrico.

OBJECTIVE: To study the proliferative activity of AgNORs (argyropholic nucleolar organizer region) in gastric cancer tissue and in the adjacent normal tissue and correlate with the clinical and pathological parameters. MATERIAL AND METHODS: Thirty four formalin-fixed paraffin-embedded tissues sections of patients with adenocarcinoma of the stomach were analysed retrospectively, being 23 of intestinal and 11 of diffuse type of Lauren. The AgNORs can be detected by a specific silver staining and the number de AgNORs was counted in 100 nucleus of tumoral cells and in 100 nucleus of the adjacent normal tissue. RESULTS: The tumoral cells had numerous, smaller, aggregated or one solitary and bigger AgNORs in the nuclei, while the normal cells had one or two AgNORs by nuclei. A significant difference was found between the mean numbers of AgNORs in the nuclei of tumor (3.86) and the normal tissue (1.25). There was also a significant difference between intestinal (4.43) and diffuse type (2.58) means. No significant correlation was found between mean AgNOR count and sex or age of the patients. CONCLUSIONS: The quantification of AgNORs can be used as a marker of proliferative activity, helping on the diagnosis of cancer.

C-erbB-2 oncoprotein in gastric carcinoma: correlation with clinical stage and prognosis.

The aim of this study was to investigate the expression of the oncogene c-erbB-2 in gastric tumors. Immunohistochemical study of the expression of c-erbB-2 was performed in formalin-fixed, paraffin-embedded sections from 82 gastric adenocarcinomas using polyclonal antibody. c-erbB-2-positive immunostaining was observed in 37 (45%) tumors. Positive staining was detected in 63% of well differentiated, 46% of moderately differentiated and 80% of papillary adenocarcinomas. In poorly differentiated adenocarcinomas, positivity for c-erbB-2 was observed in 21 %. According to the Lauren classification, a higher frequency of c-erbB-2 positive staining was observed in intestinal type tumors (70%). During the follow-up period 43% of the patients with c-erbB-2 oncoprotein-negative tumors and 45% of the patients with c-erbB-2 oncoprotein-positive tumors died. There was no significant association between c-erbB-2 staining and sex, age, clinical stage, tumor grade, histological type or survival rates. In conclusion, almost half of the gastric cancers were positive for c-erbB-2. Nonetheless, the expression of c-erbB-2 oncoprotein did not play a role in prognosis.

Estrogen and progesterone receptors in gastric and colorectal cancer.

BACKGROUND/AIMS: Estrogen (ER) and progesterone receptors (PR) have been evaluated in gastrointestinal cancer by several groups with conflicting results. The aim of the study is to examine the presence of these receptors in gastric and colon cancer. METHODOLOGY: Estrogen (ER) and progesterone (PR) receptors were assayed by the dextran-coated charcoal adsorption method from malignant and normal adjacent tissues in 16 patients with gastric adenocarcinomas and in 10 with colorectal adenocarcinomas. RESULTS: In gastric cancer, ER were detected in 62.5% and PR in 75% of the patients. In colorectal cancer, the ER and PR were detected in 60% of the patients. The binding activity ranged from 1.14-9.27 fmol/mg protein for estradiol and from 1.43-10.84 fmol/mg protein for progesterone. ER and PR were detected in normal gastric tissue in 62.5% and in 50%, respectively. In the normal colorectal tissue the ER and PR were detected in 30% and 50%. ER ranged from 1.20-16.63 fmol/mg protein for estradiol and from 1.44-9.94 fmol/mg protein for progesterone. There was no statistical difference in levels of ER and PR in both tissues. CONCLUSIONS: ER and PR were detected in normal and cancer tissues in low levels, suggesting a feature of the tissue rather than a consequence of a malignant process. Eventual role of ER or PR in these cancers remains to be elucidated.

c-erbB-2 DNA amplification and mRNA expression in human primary breast tumors and its relationship to other prognostic factors.

In order to evaluate the involvement of cerbB-2 oncogene in the etiology and progression of breast cancer, DNA samples from 157 primary human mammary carcinomas were subjected to Southern and dot blot analyses for the presence of c-erbB-2 protooncogene alterations. None of 157 carcinomas analyzed showed c-erbB-2 rearrangement. Amplification of the c-erbB-2 was found in 28.6% (45/157) of the samples. Gene expression could be analyzed in only 97 of these tumors. High levels of c-erbB-2 transcripts were detected in 25.7% (25/97) of the tumor RNA preparations. Although there was a good correlation between c-erbB-2 overexpression and gene amplification, a significant proportion of the tumors showed overexpression in the absence of gene amplification or gene amplification without overexpression. A significant correlation between c-erbB-2 proto-oncogene alterations with the number of positive lymph nodes and tumor necrosis was found, suggesting that c-erbB-2 genetic alterations have a biological importance in the etiology of breast cancer.

Association of genetic alterations of c-myc, c-fos, and c-Ha-ras proto-oncogenes in colorectal tumors. Frequency and clinical significance.

Using Northern and dot-blot analysis we examined normal and tumor tissue from 29 patients with colorectal carcinomas for the expression and amplification of c-myc, c-fos and c-Ha-ras proto-oncogenes. Overexpression of c-myc (6/24), c-fos (4/24), and c-Ha-ras (9/23) was found. For the c-fos proto-oncogene we also have observed decreased levels of expression in 13 percent (3/24) of the cases analyzed. Gene amplification appeared to be a rare event in these tumors and was found in 3/29 (10 percent) tumors for c-myc and in 1/29 (3 percent) for c-fos proto-oncogene. Curves for overall survival and for disease-free survival failed to show a significant tendency in these parameters to be poorer in tumors with alterations of gene expression for any of the proto-oncogenes analyzed. Despite the biologic importance of these genetic alterations in the etiology of colorectal tumors, levels of c-myc, c-fos, and c-Ha-ras gene expression separately or together cannot be considered as prognostic factors for clinical outcome of the disease.

Glucocorticoid receptor in active pulmonary sarcoidosis.

Glucorticoid receptor (GR) content of open-chest lung biopsies of 8 patients with active non treated pulmonary sarcoidosis was compared to 10 normal lungs. Results showed a higher GR level in sarcoid lungs (86.0 +/- 8.0 fMol/mg protein) than in control ones (8.2 +/- 3.1 fMol/mg protein). Our findings suggest that this increase is due to the accumulation of active cells in the sarcoid granuloma, possibly lymphocytes and or macrophages that may express cytosolic GR. The high level of this receptor in sarcoidosis may represent an indicator of the responsiveness of this disease to corticoids.

Multiple steroid receptors in nasopharyngeal angiofibromas.

Cytosols from 12 nasopharyngeal angiofibroma tissues were analyzed for the presence of estrogen (ER), progesterone (PR), androgen (AR), and glucocorticoid (GR) receptors, by using a dextran charcoal assay. Progesterone receptors (58%) clearly predominate over ER and AR (25%). Thus, nasoangiofibromas can be classified as ER-PR+ tumors. Glucocorticoid receptors were found in 84% of the tumors. Steroid receptor levels, although slightly higher than those determined in three specimens of normal turbinate, were quantitatively low. Positive PR and AR tumors appeared to correlate with high density of fibroblasts and endothelial cells.