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.

First molecular detection and characterization of Akabane virus in small ruminants in Turkey.

Abortion outbreaks associated with congenital malformations in two distinct small-ruminant flocks were reported in Turkey in 2013-2014. This paper describes the first molecular characterization of Turkish Akabane virus strains in small-ruminant flocks using partial sequence analysis of the S segment and pathological findings.

West Nile Virus Infection in Horses: Detection by Immunohistochemistry, In Situ Hybridization, and ELISA.

This study describes the clinicopathologic findings in naturally occurring West Nile virus (WNV) infection in horses. WNV was diagnosed in a foal by immunohistochemical and in situ hybridization methods, and the presence of WNV antibodies was detected in 5 other horses with clinical signs suggestive of WNV infection. At necropsy of the foal, lymph nodes were edematous and enlarged, and the intestines showed diffuse congestion and focal hemorrhages. The most significant histologic lesions in this case were nonsuppurative meningoencephalomyelitis, particularly in the brainstem and spinal cord. Identification of viral RNA by in situ hybridization and viral antigen by immunohistochemistry was concentrated primarily in nerve fibers, glial cells, and their processes in brainstem and spinal cord and, to a lesser extent, within the cerebral hemispheres and cerebellum.

Experimental Lactococcus garvieae infection in rainbow trout, Oncorhynchus mykiss, Walbaum 1792: a comparative histopathological and immunohistochemical study.

The aim of this study was to induce Lactococcus garvieae infection in young and adult fish through different routes [intraperitoneal (IP) and immersion (IM)] and to investigate the pathogenesis and histopathological and immunohistochemical findings comparatively. For this purpose, a total of 180 rainbow trout (90 young, 20 ± 5 g and 90 adult, 80 ± 10 g) obtained from a commercial fish farm were used. The fish were divided into eight groups, four experimental groups (Young-Adult IP groups and Young-Adult IM groups, each contain 30 fish) and four control groups (Young-Adult IP Control groups and Young-Adult IM control groups, each contain 15 fishes). The experimental study was conducted using L. garvieae, and confirmatory identification was performed by PCR. The sequence result of the PCR amplicon of 16S rDNA from isolate L. garvieae LAC1 was determined and deposited in the GenBank database under accession number KC883976. Fish in the IP groups were intraperitoneally administered an inoculate containing 10(6) cfu mL(-1) bacteria 0.1 mL. In the IM groups, fish were kept in inoculated water containing 10(8) cfu mL(-1) bacteria for 20 min. Mortality as well as clinical and pathological findings was recorded daily, and significant differences in macroscopic and microscopic results were observed between the IP and IM administration groups. All tissue samples were immunohistochemically stained by the avidin-biotin-peroxidase complex and immunofluorescence (IF) methods using polyclonal antibody to detect L. garvieae antigens. In immunoperoxidase staining in the IP groups, positive reactions to bacterial antigens were most commonly seen in the spleen, kidney, heart, liver, peritoneum and swim bladder. In the IM groups, bacterial antigens were most commonly found in the eye, gill, spleen and kidney. In the IF method, the distribution of antigens in tissue and organs was similar to the reactions with immunoperoxidase staining. Finally, in this experimental study, an important correlation was seen between the distribution of L. garvieae antigens and lesions developing in many organ and tissues.

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.

Neuropathologic study of border disease virus in naturally infected fetal and neonatal small ruminants and its association with apoptosis.

The present study describes the pathologic changes and cellular apoptosis in the central nervous system (CNS) of fetal and neonatal small ruminants infected with border disease virus (BDV), as demonstrated by immunohistochemistry and in situ hybridization. Abortions of ewes and goats were observed, as were births of lambs and kids with poor survival rates and nervous signs. Lesions included cerebellar hypoplasia, porencephaly, hydranencephaly, and nonsuppurative meningoencephalomyelitis with hypomyelinogenesis. Viral antigens and RNA were present in neuropil, glial, and neuronal cells, especially in periventricular areas, cerebellum, and brainstem. TUNEL positivity and labeling of anti-bax and anti-caspases 3, 8, and 9 were detected in BDV-infected CNSs, especially in glial and neuronal cells. The double immunostaining and TUNEL assay revealed that in BDV-infected animals, not only were BDV-infected glial and neuronal cells undergoing apoptosis, but so were uninfected cells in close vicinity of BDV-infected cells. The expression of activated caspases 3, 8, 9; bax; and TUNEL in glial and neuronal cells of the infected fetal and neonatal kids were significantly (P < .05) higher than those of the infected fetal and neonatal lambs. Yet, the expression of bcl-2 in the CNSs of the infected fetal and neonatal lambs was higher (P < .05) in neuronal and glial cells than in those of the infected fetal and neonatal kids. The results suggest that cell death in the BDV-infected CNS is induced by intrinsic and extrinsic cascades of apoptotic pathways.