Animal model for age- and sex-related genotoxicity of diethylstilbestrol

Environmental xenoestrogens pose a significant health risk for all living organisms. There is growing evidence concerning the different susceptibility to xenoestrogens of developing and adult organisms, but little is known about their genotoxicity in pre-pubertal mammals. In the present study, we developed an animal model to test the sex- and age-specific genotoxicity of the synthetic estrogen diethylstilbestrol (DES) on the reticulocytes of 3-week-old pre-pubertal and 12-week-old adult BALB/CJ mice using the in vivo micronucleus (MN) assay. DES was administered intraperitoneally at doses of 0.05, 0.5, and 5 µg/kg for 3 days and animals were sampled 48, 72 and 96 h, and 2 weeks after exposure. Five animals were analyzed for each dose, sex, and age group. After the DES dose of 0.05 µg/kg, pre-pubertal mice showed a significant increase in MN frequency (P < 0.001), while adults continued to show reference values (5.3 vs 1.0 MN/1000 reticulocytes). At doses of 0.5 and 5 µg/kg, MN frequency significantly increased in both age groups. In pre-pubertal male animals, MN frequency remained above reference values for 2 weeks after exposure. Our animal model for pre-pubertal genotoxicity assessment using the in vivo MN assay proved to be sensitive enough to distinguish age and sex differences in genome damage caused by DES. This synthetic estrogen was found to be more genotoxic in pre-pubertal mice, males in particular. Our results are relevant for future investigations and the preparation of legislation for drugs and environmentally emitted agents, which should incorporate specific age and gender susceptibility.

Animal model for age- and sex-related genotoxicity of diethylstilbestrol.

Environmental xenoestrogens pose a significant health risk for all living organisms. There is growing evidence concerning the different susceptibility to xenoestrogens of developing and adult organisms, but little is known about their genotoxicity in pre-pubertal mammals. In the present study, we developed an animal model to test the sex- and age-specific genotoxicity of the synthetic estrogen diethylstilbestrol (DES) on the reticulocytes of 3-week-old pre-pubertal and 12-week-old adult BALB/CJ mice using the in vivo micronucleus (MN) assay. DES was administered intraperitoneally at doses of 0.05, 0.5, and 5 microg/kg for 3 days and animals were sampled 48, 72 and 96 h, and 2 weeks after exposure. Five animals were analyzed for each dose, sex, and age group. After the DES dose of 0.05 microg/kg, pre-pubertal mice showed a significant increase in MN frequency (P < 0.001), while adults continued to show reference values (5.3 vs 1.0 MN/1000 reticulocytes). At doses of 0.5 and 5 microg/kg, MN frequency significantly increased in both age groups. In pre-pubertal male animals, MN frequency remained above reference values for 2 weeks after exposure. Our animal model for pre-pubertal genotoxicity assessment using the in vivo MN assay proved to be sensitive enough to distinguish age and sex differences in genome damage caused by DES. This synthetic estrogen was found to be more genotoxic in pre-pubertal mice, males in particular. Our results are relevant for future investigations and the preparation of legislation for drugs and environmentally emitted agents, which should incorporate specific age and gender susceptibility.

[Dermatomyositis and malignant disease--case report]. / Dermatomiozitis i maligna bolest--prikaz bolesnika.

The case of patients with dermatomyositis and malignant disease has been described. The diagnosis of malignant disease was made after death of the patient, when the malignant undifferentiated cells were found in the ascites. We think that dermatomyosistis was caused by malignant disease. The similar cases of disease were described and published in the literature.

Expression of erbB-3 protein in colorectal adenocarcinoma: correlation with poor survival.

BACKGROUND/AIMS: The family of erbB receptors includes four transmembrane glycoproteins with tyrosine kinase activity. These receptors are widely expressed in normal tissues, but they also have been implicated in the development of several human adenocarcinomas. c-erbB-3/HER-3 has been detected to a greater or lesser extent in many tissues from the digestive, urinary, reproductive and respiratory tracts. The overexpression of c-erbB-3/HER-3 protein has also been shown in 53%-88% of colorectal adenocarcinomas. In this study we investigated the expression of the c-erbB-3/ HER-3 gene product in colorectal tumour samples, and compared the results obtained with several clinicopathological parameters, including the survival of patients. METHODS: Paraffin-embedded tissue sections were analysed immunohistochemically, using monoclonal antibody RTJ1 to human erbB-3 protein. Antibody RTJ1 specificity was confirmed by immunoprecipitation followed by Western blotting analysis. Amplification of the erbB-3 oncogene was tested by dot-blot hybridization. RESULTS: Adenocarcinomas of the colon were positive for erbB-3 protein in 78% of samples examined. Dot-blot analysis showed no amplification of the erbB-3 gene in colon adenocarcinomas. Statistical analysis showed that patients with tumours that could not be stained for erbB-3 protein survived significantly longer (P<0.05) than patients with tumours staining positive for the erbB-3 protein. A Cox proportional-hazards model with stepwise variable selection identified age, sex and erbB-3 expression as important prognostic factors. CONCLUSION: These findings demonstrate that erbB-3 protein expression could serve as a prognostic factor in colorectal malignancies.

Dental amalgam mercury exposure in rats.

The aim of this study was to measure the distribution of mercury, in tissues of rats exposed to amalgam over a two months period. Possible interaction of mercury with copper and zinc in organs was also evaluated. Rats were either exposed to mercury from 4 dental amalgams, or fed the diet containing powdered amalgam during two months. Mercury was measured in the kidney, liver and brain, copper in kidney and brain and zinc in kidney. The results showed significantly higher concentrations of mercury in the kidneys and the brains of rats in both exposed groups compared to control. Even after two months of exposure to mercury brain mercury concentration in rats with amalgam fillings was 8 times higher than in the control and 2 times higher than in rats exposed to amalgam supplemented diet. The highest mercury concentration in the latter group was found in the kidneys and it was 5 times higher than in the control group. We found no significant differences between mercury levels in exposed and control rat's liver. Exposure to mercury from dental amalgams did not alter the concentrations of copper and zinc in the tissues. Histopathological analyses of rats tissues did not show any pathological changes. These results support previously proposed nose-brain transport of mercury released from dental amalgam fillings.

Effect of cromolyn pre-treatment on capsaicin-induced rhinitis in rats.

The effect of cromolyn sodium local pre-treatment on capsaicin-induced rhinitis in rats was studied by analyzing tissue changes due to edema, inflammatory cell infiltration and IgA upregulation. Nasal mucosa samples were stained with hematoxylin-eosin and examined immunohistochemically with monoclonal IgA antibodies. Changes were analyzed at 6, 12 and 72 h after capsaicin treatment and were scored semiquantitatively. Results showed that local cromolyn pre-treatment modified all parameters observed in the nasal mucosa following capsaicin-induced rhinitis in the rats.

Apoptosis in human gastric polyps and adenocarcinomas: a stereological analysis.

BACKGROUND/AIM: The occurrence of apoptotic cells was analyzed in human normal gastric mucosa, polyps and adenocarcinomas. METHODOLOGY: Histological classification was carried out on hematoxylin and eosin stained slides. The tissue was classified as follows: Normal gastric mucosa or adenomatous polyps. Gastric carcinoma specimens were histologically classified according to Lauren's classification into: A: Diffuse adenocarcinoma without metastasis, B: Diffuse adenocarcinoma with metastasis, C: Intestinal adenocarcinoma without metastasis, D: Intestinal adenocarcinoma with metastasis, E: Mixed adenocarcinoma without metastasis and mixed adenocarcinoma with metastasis. The counting of apoptotic cells was performed using the 40X objective with a calibrated eyepiece Weibel's multi-purpose M 42 stereological test system. Each group was evaluated stereologically, determining numeric density of apoptotic cells. RESULTS: The results show the progressive and statistically significant increase of apoptotic numeric densities from normal gastric epithelium to adenomatous polyp and finally to cancer, which contained the highest number of apoptotic cells. Comparing gastric carcinoma with and without metastasis in intestinal and diffuse adenocarcinoma there was statistically significant difference. In these two groups, carcinomas with metastasis contained higher number of apoptotic cells than without metastasis. Gastric cancer according to numeric densities of apoptotic cells can be separated in tree statistically different groups: A: Intestinal type gastric cancer with metastasis (the highest number of apoptotic cells), B: Intestinal type gastric cancer without metastasis and diffuse gastric cancer with metastasis (medium number), C: Diffuse type gastric cancer without metastasis, mixed gastric cancer with and without metastasis (the lowest number of apoptotic cells). CONCLUSION: These results suggest that numeric densities of apoptotic cells are associated with tumor progression in human gastric carcinogenesis and can be used as prognostic mark.

The expression of p185(HER-2/neu) correlates with the stage of disease and survival in colorectal cancer.

BACKGROUND & AIMS: HER-2/neu oncogene encodes a transmembrane tyrosine kinase receptor that is amplified and/or overexpressed predominantly in adenocarcinomas. This phenomenon has been most intensively studied in breast carcinoma where its amplification and overexpression correlate with the overall course of disease and poor prognosis. This study was designed to investigate HER-2/neu gene expression in benign and malignant colorectal lesions and to evaluate its prognostic importance in colorectal cancer. METHODS: Two hundred twenty-one samples of normal colon, benign lesions, and colorectal adenocarcinomas were studied for expression of HER-2/neu oncoprotein. Immunohistochemical staining of formalin-fixed, paraffin-embedded tissue sections of primary tumor and lymph nodes was performed. Immunoprecipitation followed by Western blotting of freshly frozen samples of the same tumors were also performed. RESULTS: Normal colon mucosa, benign lesions, and adenocarcinomas clearly differed in the expression levels and histological distribution of p185(HER-2/neu). Normal mucosa was mostly negative, but significant number of benign lesions and adenocarcinomas overexpressed HER-2/neu protein. Adenocarcinomas were significantly more positive than benign lesions. The results show significant correlation with the epithelial abnormality degree and clinical parameters including Dukes' classification and relapse-free and postoperative survival period. CONCLUSIONS: The p185(HER-2/neu) rate expression could serve as an independent prognostic factor in patients with p185(HER-2/neu)-positive colorectal malignancies.

Adrenal cavernous hemangioma: MRI, CT, and US appearance.

Two cases of rare adrenal cavernous hemangiomas are reported, one imaged with conventional X-ray techniques, US, CT, and MRI, and the other with US and CT. The CT technique clearly demonstrated calcifications and the internal structure of the lesions in both cases and peripheral rim enhancement on the postcontrast scan in one patient. Although MRI demonstrated accurately the complex nature of the lesion, the inability to visualize the calcified areas do not allow to make a specific histologic diagnosis.

Pulmonary blastoma with fatal hypercalcemia.

We describe a 50-year-old, previously healthy male with metastatic pulmonary blastoma associated with hypercalcemic and hyperosmolar complications which caused his death after 5 days. The primary tumor consisted of epithelial [cytokeratin, beta-hCG CEA, neuron-specific enolase (NSE)-positive] and mesenchymal components (beta-hCG, vimentin, NSE-positive, while the metastases had only a mesenchymal component.

Dose-dependent protective effect of BPC 157 on capsaicin-induced rhinitis in rats.

Protection of BPC 157 on capsaicin-induced rhinitis was studied in Wistar rats for its effect on mastocyte infiltration, degranulation and inflammatory cell infiltration. Animals were pretreated with 10 microg/kg, 10 ng/kg or 2 ml saline i.p. and capsaicin (0.05 ml/nostril of 1750 nmol/l sol.) was applied intranasally. They were then euthanized at 1, 3 and 12 h after capsaicin provocation. Nasal mucosa was analyzed and scored for mastocyte infiltration, degranulation and inflammatory cell infiltration. BPC 157 pretreatment significantly prevented mastocyte infiltration at 1 h. Polymorphonuclear leukocyte infiltration was significantly reduced in rats pretreated with 10 microg/kg BPC 157. A dose-dependent effect of BPC 157 pretreatment was demonstrated only for polymorphonuclear leukocyte infiltration at 12 h.

A female patient with tuberculous polyserositis, and congenital tuberculosis in her new-born child.

A 24-year-old woman was admitted to our department with sepsis, polyserositis and suspected abdominal tumor immediately after delivery. Effusion analyses suggested an etiology of tuberculosis. The severe general condition of the patient allowed for no invasive diagnostic procedures. Antituberculosis therapy was administrated too late and was of no benefit. The fatal outcome was due to septic shock and adult respiratory distress syndrome (ARDS). Autopsy findings confirmed tuberculous polyserositis as well as tuberculous infection of the lungs. The same diagnosis was established in her newborn infant, who responded well to antituberculosis treatment.

[Interferon in the treatment of solid tumors]. / Interferon u lijecenju solidnih tumora.

Discovered almost four decades ago, interferons were being investigated as potential anticancer agents from early sixties, because of their antiviral antiproliferative and immunomodulating properties and their relatively modest toxicity. Unlike very good effects on some lymphoproliferative disorders where complete durable remissions were observed, results of their application in solid tumors are not so unambiguous. Among large number of tumors investigated so far, the best, reproducible, although modest, activity was found against melanoma, renal cell carcinoma and Kaposi's sarcoma. There are still many doubts concerning their optimal doses, sequencing and schedules, most probably in combination with other biological response modifiers and/or cytotoxic agents, which have to be clarified in further clinical trials, together with investigation of more reliable biological indicators of response to interferons in vivo.

Pheochromocytoma without hypertension.

Pheochromocytoma is a rare tumour which is usually suggested by sustained or paroxysmal hypertension. Our patient with a pheochromocytoma was unusual for two reasons: she was normotensive during 3 weeks of close observation in the hospital and the urinary adrenaline level was higher than the noradrenaline level. In the patient's history there were no data on hypertension attacks. The patient had a palpable abdominal mass and the cytological finding was in contrast with a clinical diagnosis of pheochromocytoma. The surgery revealed a tumour and the pathohistologic evaluation established the diagnosis of pheochromocytoma. After surgery, the patient's urinary catecholamine levels reverted to normal. Our patient may have been normotensive because her tumour predominantly secreted adrenaline and perhaps other vasodilating substances.

Autopsy and the internal medicine department.

The accuracy of major clinical diagnoses and causes of death was estimated from the analysis of clinical diagnoses and autopsy findings of patients deceased at the internal medicine department of a large university hospital in two different periods (1965-1967 and 1985-1987). The autopsy cases from the second period were subdivided in two groups i.e. less and more than seven days of hospital stay prior death. From 100 randomly selected autopsy cases in each period there was no statistically significant variability in clinico-pathological concordance. In three groups studied (1965-1967, 1985-1987 < seven days and 1985-1987 < seven days hospital stay) the major clinical diagnosis was confirmed in 85, 94 and 86% respectively. A decrease of nearly 27% in the number of autopsies performed is shown between analysed periods. Clinico-pathological conferences are regularly held weekly, this being one of the reasons for a stable and extremely high clinico-pathological concordance.

Autopsy and clinical diagnoses in a psychiatric hospital.

We made a retrospective analysis of clinical diagnoses and autopsy findings for patients who died in the psychiatric hospital in the city of Zagreb in two different periods--1965-67 and 1985-87. Between these two periods, the autopsy rate decreased from 64 to 31%. The level of clinicopathological discrepancies remained the same, but with the decrease in autopsy rate a problem emerged in the diagnosis of respiratory diseases. The purpose and value of autopsy in a psychiatric hospital is discussed.