Funda Yildirim

Molecular and pathological investigations of EHV-1 and EHV-4 infections in horses in Turkey

The aim of the present study was to investigate abortion storms that occurred in the Marmara region of Turkey in 2008-2009 using a real-time PCR. Two aborted foetuses were necropsied and histo-pathological findings reported herein. Ten lungs, 3 brains and one nasal swab from 10 aborted foetuses, 6 nasal swabs and 3 vaginal swabs from aborting mares were included in this study. EHV-1 was isolated from the lung, liver and brain of 1 aborted foetus. EHV-1 DNA was detected in the lungs, livers and spleens of 2 necropsied foetuses and in 3 lungs from 10 foetuses submitted for diagnosis. A brain from one of the aborted foetuses was also positive for EHV-1 DNA. EHV-4 DNA was detected only in a nasal swab of one of the tested foetuses. Neither EHV-1 nor EHV-4 DNA was detected in the swabs of aborting mares. Sequence analysis of the glycoprotein B of the strains was performed and a phylogenetic tree was generated. The results indicated that 4 of the 5 Turkish EHV-1 strains (TR02, TR03, TR04 and TR05) clustered together; the fifth strain (TR01) was slightly removed from the group and clustered with other EHV-1 from various origins. Single nucleotide polyporphism (SNP in ORF30) associated with neuropathogenesis was not detected in any of the strains. At necropsy, sub-milier focal necrosis in the liver and spleen was observed. Microscopically, focal coagulation necrosis and marked eosinophilic intranuclear and intracytoplasmic inclusion bodies in the hepatocytes localised around the necrotic areas in the liver. Severe coagulation necrosis in white pulp of the spleen was also observed.

Is Chronic Curcumin Supplementation Neuroprotective Against Ischemia For Antioxidant Activity, Neurologic Deficit Score, Or Neuronal Apoptosis In An Experimental Stroke Model?

AIM To investigate the neuroprotective effect of chronic curcumin supplementation on the rat forebrain prior to ischemia and reperfusion. MATERIAL AND METHODS Forebrain ischemia was induced by bilateral common carotid artery occlusion for 1/2 hour, followed by reperfusion for 72 hours. Older rats were divided into five groups: Group I received 300 mg/kg oral curcumin for 21 days before ischemia and 300 mg/kg intraperitoneal curcumin after ischemia; Group II received 300 mg/kg intraperitoneal curcumin after ischemia; Group III received 300 mg/kg oral curcumin for 21 days before ischemia; Group IV had only ischemia; Group V was the sham-operated group. The forebrain was rapidly dissected for biochemical parameter assessment and histopathological examination. RESULTS In forebrain tissue, enzyme activities of superoxide dismutase, glutathione peroxidase, and catalase were significantly higher in Group I than Groups II or III (p < 0.05) while xanthine dehydrogenase and malondialdehyde enzyme activities and concentrations of interleukin-6 and TNF-alpha were significantly lower in Group I when compared to Groups II and III (p < 0.05). A significant reduction in neurological score was observed after 24 and 72 hours in the curcumin-treated groups compared with the ischemic group. We also found a marked reduction in apoptotic index after 72 hours in the groups receiving curcumin. Significantly more TUNEL-positive cells were observed in the ischemic group compared to those treated with curcumin. CONCLUSION We demonstrated the neuroprotective effect of chronic curcumin supplement on biochemical parameters, neurological scores and apoptosis following ischemia and reperfusion injury in rats.

Synergistic growth inhibitory effect of deracoxib with doxorubicin against a canine mammary tumor cell line, CMT-U27.

Cyclooxygenase (COX) inhibitors have been shown to exert anti-angiogenic and anti-tumor activities on many types of malignant tumors. These anticancer properties make it worthwhile to examine the possible benefit of combining COX inhibitors with other anti-cancer agents. In the present study, we evaluated the potential of deracoxib (DER) in potentiating antitumor activity of doxorubicin (DOX) in canine mammary carcinoma cells (CMT-U27). DER (50–250 µM) enhanced the antiproliferative activity of DOX by reducing the IC50 (approximately 3- to 3.5 fold). Interaction analysis of the data showed that combinations of DOX at 0.9 µM with DER (100–250 µM) produced synergism in the CMT-U27 cell line, with a ratio index ranging from 1.98 to 2.33. In additional studies identifying the mechanism of observed synergistic effect, we found that DER strongly potentiated DOX-caused G0/G1 arrest in cell cycle progression. Also, DER (100–250 µM) augmented apoptosis induction with approximately 1.35- and 1.37- fold increases in apoptotic response caused by DOX in the cells. DER enhanced the antiproliferative effect of DOX in conjunction with induction of apoptosis by modulation of Bcl-2 expression and changes in the cell cycle of the CMT-U27 cell line. Although the exact molecular mechanism of the alterations in the cell cycle and apoptosis observed with DER and DOX combinations require further investigations, the results suggest that the synergistic effect of DOX and DER combinations in CMT therapy may be achieved at relatively lower doses of DOX with lesser side effects. Therefore, combining DER with DOX may prove beneficial in the clinical treatment of canine mammary cancer.

Clinical, Virological and Pathological Investigations on Horses with Neurological Disorders in Turkey

ABSTRACT The aim of this study was to investigate the clinical, pathologic, and viral etiology of horses with neurologic disorders. Twelve English Thoroughbred horses with neurologic disorders were investigated for the presence of neuropathogenic equine herpesvirus‐1 (EHV‐1) and Borna disease virus (BDV) by real‐time polymerase chain reaction (PCR) and histopathological methods. Neuropathogenic EHV‐1 was detected in the brain of two horses by real‐time PCR. Borna disease virus p24 and p40 gene sequences were detected by real‐time RT‐PCR in the brain of a 3‐year‐old horse and in the blood of a 1‐year‐old horse. High fever, ataxia, depression, lack of coordination, and gait abnormalities were present in these horses, which died within a few days of developing neurologic signs. The BDV p24 and p40 real‐time PCR products were sequenced and shown to be identical to previously reported BDV sequences. In the brain of the BDV‐positive horse, hyperemia was pronounced in the parenchyma and the meninges. In addition, nonpurulent poliencephalomyelitis characterized by perivascular mononuclear cell infiltration was seen. Mononuclear and polymorphonuclear cell infiltration was also seen in the liver with necrotic perihepatitis besides subacute splenitis and glomerulonephritis and severe hyperemia in the kidney and edema and emphysema in the lung. In conclusion, neuropathogenic EHV‐1 and BDV were detected in horses from Turkey using molecular detection methods. Since BDV genetic signatures were detected in Turkey for the first time, future epidemiologic studies need to be performed to investigate the range of host animals, spread, frequency, and molecular diversity of BDV; this will allow to determine the risk of this pathogen for Turkish veterinary and public health. HighlightsThis study describes the first cases of Borna disease in horses and first molecular detection of Borna disease virus in Turkey.

Investigation of human papillomaviruses (HPV), mouse mammary tumour virus (MMTV), Epstein–Barr virus (EBV), and human polyomavirus entities in canine mammary tumours

Abstract Introduction: The aim of the study was to investigate the presence of human papillomaviruses (HPV), mouse mammary tumour virus (MMTV), Epstein-Barr virus (EBV), and human polyomavirus BK in canine mammary tumours (CMTs) and to correlate the results of histopathological classification with the results of virological examination. Material and Methods: Eighty CMTs and ten normal canine mammary gland samples were evaluated using histopathological methods and TaqMan real-time PCR analysis. Results: The results indicated that all mammary tumours and normal mammary tissue samples were negative for HPV16 and other HPV, EBV, human polyomavirus, and human mammary tumour virus strains. Conclusion: Further studies should be performed to investigate the existence of other strains of HPV, EBV, and human polyomavirus in CMTs.

Immunohistochemical detection of tumour cell proliferation and intratumoural microvessel density in canine malignant mammary tumours

Abstract The objective of this study was to investigate the correlation between different histological types and grades of canine malignant mammary tumours, tumour cell proliferation and their angiogenic activity using immunohistochemical markers. Mammary tissue samples from 47 bitches with mammary cancer were evaluated. The expression of cellular proliferation marker Ki-67 and endothelial marker Von Willebrand’s factor (vWF) were immunohistochemically demonstrated. The tumours with the highest Ki-67 and vWF expressions were found to share similar histomorphological features. Simple solid carcinoma had the highest levels of Ki-67, vWF, and higher histological grade while complex carcinomas, osteosarcomas, and carcinosarcomas had the lowest ones. The differences between the expressions of Ki-67 and vWF in different tumour types were considered to be of great importance in determination of biological behaviour and prognosis of these tumours. This study is one of the few studies that evaluate these differences among the subtypes of malignant canine mammary tumours