Protective effects of silymarin and curcumin on cyclophosphamide-induced cardiotoxicity.

INTRODUCTION: Cyclophosphamide (CP) is a potent anticancer agent; its clinical use is limited due to its marked cardiotoxicity. AIM: The present study was aimed at evaluating the cardioprotective effects of silymarin (SLY) and curcumin (CUR), which have strong antioxidant properties, against the toxic effects of high-dose CP on the heart of rats. MATERIALS AND METHODS: A total of 36 adult Wistar albino female rats were randomly divided into six groups. Group I (control group; nothing was administered), Group II (CP group; 30mg/kg/day CP was administered intraperitoneally to each animal for seven days), Group III (SLY group; 100mg/kg/day SLY by gavage for 14 days), Group IV (CUR group; 100mg/kg/day CUR by gavage for 14 days), Group V (SLY+CP group; 100mg/kg/day SLY by gavage for 14days plus 30mg/kg/day CP intraperitoneally starting from the seventh day) and Group VI (CUR+CP group; 100mg/kg/day CUR by gavage for 14days plus 30mg/kg/day CP intraperitoneally starting from the seventh day). Biochemical, histopathological and immunohistochemical methods were utilised for evaluation of the cardiotoxicity. RESULTS: The result showed that an increase in heart MDA and DNA fragmentation levels were detected while significant decreases were seen in SOD levels in CP alone group when compared to the other groups. CP caused severe damage in the histopathological status of heart tissue including intersititial oedema, haemorrhage, degeneration and necrosis in muscle fibrils and perinuclear vacuolization. A significant increase in the percentage of TUNEL-positive cells and γH2AX protein expression was detected in the CP-treated group compared to the control and other treated groups. There was significant increase in the percentage of caspase 3-positive cells and decrease in the percentage of Bcl-2 positive cells in the CP group compared to the control group and other treated groups. However, a significant decrease in the percentage of cTnI and cTnT immunoreactivity was also observed in the CP-treated group compared to the control and other treated groups. In the groups in which SLY and CUR were administered concurrently with CP, biochemical parameters, histopathological and immunohistochemical results were found to be significantly lower than in the CP-only group. CONCLUSIONS: These results lead to conclusion that the natural antioxidant SLY and CUR might have protective effects against CP-induced cardiotoxicity and oxidative stress in rats.

Pathomorphological, immunohistochemical and bacteriological findings in budgerigars (Melopsittacus undulatus) naturally infected with S. Gallinarum.

The present study describes the pathological and bacteriological findings and diagnosis by immunoperoxidase and immunofluorescence methods in budgerigars (Melopsittacus undulatus) naturally infected with Salmonella gallinarum obtained from three commercial budgerigar rearing farms. The course of the disease in young budgerigars was peracute or acute, whereas in adult budgerigars the disease was acute or chronic. Clinically, yellow-white diarrhoea was observed in the young budgerigars with the acute form. In the adult budgerigars with the acute and chronic forms, a decrease in feed and water consumption with loss in body condition together with greenish-yellow diarrhoea was generally noted. Peritonitis and pericarditis were the most common findings in young budgerigars at necropsy, while in adult budgerigars scattered grey-white necrotic foci were found in the livers. Histopathologically, the lesions in young budgerigars were characterized with fibrinonecrotic peritonitis and/or pericarditis and necrotic hepatitis. In adult budgerigars with acute infection, hepatic necrosis with focal heterophil infiltration was present; whilst lesions in the chronic cases were granulomatous in nature with the infiltration of macrophages, lymphocytes and histiocytes. For the detection of S. Gallinarum in formalin-fixed, paraffin-embedded tissues, the avidin-biotin peroxidase complex and immunofluorescence methods were used. Both methods showed bacteria to be localized in the liver, kidney, peritoneum, heart, spleen and intestines of both young and adult budgerigars. The results of the present study indicate that the avidin-biotin peroxidase complex method was more sensitive than the immunofluorescence method in the detection of the bacteria.

Cyclin A expression is associated with apoptosis and mitosis in murine 3-methylcholanthrene-induced fibrosarcomas.

The chemical carcinogen MCA induces fibrosarcoma and tissue damage at the injection site. Despite the importance of ROS in the development of cancer, little is known about the pattern of expression of ROS in MCA-induced fibrosarcomas. To gain some insight into the biological significance of iNOS and Cu/Zn-SOD, comparative immunohistochemical analyses were performed to characterize their expression in MCA-induced fibrosarcomas. Cyclin A is overexpressed in various tumors, but its expression in MCA-induced fibrosarcoma in mice and its correlation to mitosis and apoptosis are unclear. The presence of apoptotic cell death was evaluated using the TUNEL method and findings were compared with cyclin A expression and mitotic count of fibrosarcomas. Subcutaneous application of MCA caused fibrosarcoma development in 14 of 20 mice (70%) in 26 weeks. Limited cytoplasmic Cu/Zn-SOD and iNOS immunostainings were detected in 13 of 14 and 9 of 14 tumors with median immunoreactive scores of 2 and 1, respectively. Prominent nuclear cyclin A immunostaining and TUNEL-positive reactions were seen in all the fibrosarcoma cases. Cyclin A immunoreaction significantly correlated with the TUNEL index (P<0.01) and MC (P<0.001). The present findings show a low level of iNOS expression in neoplastic cells indicating limited synthesizing capacity of tumor cells. Limited Cu/Zn-SOD reaction could be associated with an imbalance in between pro-oxidant/antioxidant levels. Furthermore, it was shown that cyclin A is overexpressed in MCA-induced fibrosarcomas and possibly plays a significant role in the pathogenesis of fibrosarcomas. Cyclin A could be useful for detecting the S phase of the cell cycle and could also indicate that cyclin A may induce S phase arrest associated with apoptosis in the MCA-induced fibrosarcomas.

Osteoclast-like giant cell tumour arising from the kidney in a dog.

In this study, a case of osteoclast-like giant cell tumour arising from the kidney is reported in an eight-year-old female Anatolian Shepherd dog. Macroscopically, the tumorous mass covered the hilus of the left kidney. It was 26 x 22 x 12 cm in size and 3700 g in weight. Metastatic tumorous nodules, 0.5-2.0 cm in diameter, were found on the abdominal side of the diaphragm and in the lungs. Microscopically, numerous large osteoclast-like multinucleated giant cells and spindle-spheroidal-shaped cells were seen. Osteoblastic differentiation and osteoid matrix were noted in a few areas at the periphery of the tumour, near the connective tissue septa. The stroma of the tumour tissue was vascular, oedematous and loose. By immunoperoxidase staining, tumour cells showed immunoreactivity for vimentin but not for keratin and desmin, indicating that the tumour had mesenchymal origin. This is the first report in the literature on a malignant osteoclast-like giant cell tumour arising from a visceral organ in animals.

Ligand-induced expression of peroxisome proliferator-activated receptor alpha and activation of fatty acid oxidation enzymes in fatty liver.

BACKGROUND: Peroxisome proliferator-activated receptor alpha (PPARalpha) regulates lipid metabolism upon activation by ligands. Peroxisome proliferator-activated receptor alpha may play a role in the pathogenesis of fatty liver disease. The aim of this study was to assess the PPARalpha expression pattern and mitochondrial/peroxisomal enzyme activities in response to high fat diet (HFD) and clofibrate, a well known PPARalpha ligand. MATERIALS AND METHODS: Four groups of Wistar-Albino rats were included: (1) rats fed a control diet (CD) for 6 weeks, (2) rats fed CD (6 weeks) plus clofibrate (last 2 weeks), (3) rats fed HFD for 6 weeks, and (4) rats fed HFD (6 weeks) plus clofibrate (last 2 weeks). Peroxisome proliferator-activated receptor alpha expression was evaluated by immunohistochemistry. Fatty acid beta-oxidation (peroxisomal-acyl-CoA-oxidase and mitochondrial-acyl-CoA-dehydrogenase) and catalase enzyme activities, and malondialdehyde and glutathion levels were measured spectrophotometrically in liver tissues. RESULTS: All animals were fed HFD but only 2/12 animals were fed HFD plus clofibrate-developed fatty liver. Both HFD and clofibrate induced PPARalpha expression, clofibrate induction being more prominent than HFD. Clofibrate plus HFD did not further increase PPARalpha expression. Activities of peroxisomal-acyl-CoA-oxidase and mitochondrial-acyl-CoA-dehydrogenase enzymes were not induced by HFD alone. Clofibrate increased the activity of these enzymes in both CD- and HFD-fed animals. However, an increase of acyl-CoA-oxidase activity was blunted in rats fed HFD. Catalase activity and malondialdehyde levels were increased but glutathion levels were unchanged in rats fed HFD plus clofibrate. CONCLUSIONS: Clofibrate was a more potent inducer of PPARalpha expression than HFD in our rat fatty liver model. The finding of blunted peroxisomal enzyme response to clofibrate in fatty livers suggests that alterations in postreceptor events may exist and further contribute to liver steatosis. Clofibrate seems to stabilize glutathion content and this might contribute to the prevention of liver steatosis.

Electron microscopical studies on cysts of Sarcocystis arieticanis within cardiac muscle of naturally infected sheep.

Transmission electron microscopy was used to the cysts and their walls of Sarcocystis arieticanis within cardiac muscle of naturally infected sheep. The general ultrastructural features of the cysts, previously described by other authors, were confirmed. The cyst walls of S. arieticanis and the size and shape of the protrusions at different locations on the cyst wall were explained in detail. The cysts were 35-62.5 micro m x 52.5-162.5 micro m in size and the cyst wall had different-shaped protrusions. Aged cysts that were localized in the heart and differences in the morphology of their protrusions were noticed.

Generalized tuberculosis in a 45 day-old calf.

The objectives of the present study were to describe pathomorphological and immunohistochemical features of generalized natural tuberculosis in a 45 day-old female calf. The characteristic lesion of tuberculosis was productive type which was located in the lung, liver, and especially in the lymph nodes (mediastinal, bronchial, mesenterial, portal, prescapular, renal and caudal sternal lymph nodes). Mycobacterium bovis antigens were seen generally in the cytoplasma and around the macrophages, rarely in necrotic areas and Langhans giant cells by avidin-biotin complex peroxidase method.