Doğukan Özen

Ptaquiloside‑induced cytotoxicity in Crandall feline kidney and HGC‑27 cells

Ptaquiloside (PTA) is a potent genotoxic carcinogenic compound, which is found in bracken ferns and predominantly causes gastric tumors in humans, as well as bladder tumors and chronic enzootic hematuria in cattle. The underlying molecular mechanisms of PTA remain a topic for interdisciplinary investigation. The aim of the present study was to determine the possible cytotoxic effect of 24 h of PTA exposure in Crandall feline kidney (CrFK) and human gastric cells (the HGC-27 cell line) using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and lactose dehydrogenase (LDH) analysis. The cytotoxic effects of PTA (0.0005–500 μg/ml) were found to increase in a dose-dependent manner, whereby the half maximal inhibitory concentration values were 11.17 and 11.86 μg/ml in the CrFK cells, and 2.03 and 2.56 μg/ml in the HGC-27 cells, by LDH and MTT assay, respectively. The results of the present study are consistent with those of previous studies associated with the cytotoxicity of PTA; however, cytotoxicity was identified to occur at significantly lower doses. This cytotoxic effect in vitro at particularly high doses may be linked to the initiation of carcinogenesis as a result of oxidative stress.

Effect of Lonicera japonica extract on Mycoplasma gallisepticum in naturally infected broiler flocks

Abstract In this study, the effect of chlorogenic acid extract from Lonicera japonica Thunb. on Mycoplasma gallisepticum infections and the performance of broiler flocks was investigated. A total of 360 Ross-308 broiler chicks taken from M. gallisepticum seropositive flocks were divided equally into three groups designated as control (nothing administered), antibiotic (Tylosin tartrate given for the first 3 d and d 20–22) and test group (chlorogenic acid extract given twice a day on d 16 and 22). Broiler performance analysis, serological tests (slide agglutination), molecular identification (polymerase chain reaction) and histopathological examination were performed to detect M. gallisepticum. The results show that chlorogenic acid not only increases live body weight but is also an alternative treatment option in M. gallisepticum–infected broiler flocks.

Intratumoral recombinant human interferon alpha-2a and vincristine combination therapy in canine transmissible venereal tumour

Canine transmissible venereal tumour (CTVT) is a naturally occurring contagious neoplasm of dogs located mainly on the external genitalia of both sexes. The course of vincristine chemotherapy, the most effective and practical therapy, is affected by the immune status of the host. The aim was to investigate recombinant human interferon alpha-2a (rhIFNα-2a) and vincristine for treatment of CTVT. A total of 21 female dogs were included. In group I (nxa0=xa09), vincristine (0.025xa0mg/kg, IV) was administered weekly. In group II (nxa0=xa06), dogs were injected intratumorally weekly with 1.5xa0million IU rhIFNα-2a. In group III (nxa0=xa06), rhIFNα-2a and vincristine were combined. No tumour regression was observed after three injections of rhIFNα-2a in group II and weekly vincristine was administered. The number of tumour infiltrating lymphocytes (TILs), mitotic figures and apoptotic cells were counted in subsequent incisional tumour biopsies. The Kaplan-Meier Method was used to analyse survival using complete tumour regression as the outcome and Breslow Test was used for comparison of survival curves. Differences in TILs, cell proliferation and apoptosis between groups were assessed by analysis of covariance. Complete regression was observed in all animals included. Mean duration of vincristine treatment for complete regression was shorter in group II (3.50xa0weeks, 95% CI, 3.06-3.94, Pxa0<xa00.05) and group III (3.17xa0weeks, 95% CI, 2.84-3.49, Pxa0<xa00.01) compared to group I (5.11xa0weeks, 95% CI, 4.42-5.80). Vincristine and rhIFNα-2a combination increased TILs in CTVT biopsies compared to vincristine treatment (Pxa0=xa00.017) and vincristine treatment after rhIFNα-2a (Pxa0=xa00.049). Vincristine treatment after rhIFNα-2a (Group II; Pxa0<xa00.001) and rhIFNα-2a and vincristine combination (Group III; Pxa0<xa00.001) decreased apoptosis. The results indicate that intratumoral rhIFNα-2a treatment alone is not effective in CTVT. However, combination of rhIFNα-2a and vincristine shortens the duration of treatment compared to vincristine therapy.

Suitability of the hemi-zona assay for the evaluation of the effect of the length of the equilibration period before cryopreservation

The aim of this study was to test the suitability of the interspecific hemizona assay (HZA) to predict the fertilizing capacity of bovine sperm after modifying the length of the equilibration period before freezing and thawing. Ejaculates from 10 proven fertile bulls were split after dilution, equilibrated at 4xa0°C for either 24xa0h (control spermxa0=xa0CS) or 6, 48, 72 or 96xa0h (test spermxa0=xa0TS) and cryopreserved. Hemizona (HZ) pairs from inxa0vitro matured pig oocytes were used for the heterologous HZA: After thawing and swim-up (1xa0h) CS and TS were co-incubated with matching HZ (125,000xa0S/HZ in 25xa0μL Fert-TALP) for 4xa0h. Spermatological analyses (progressive motile sperm (PMS), plasma membrane- and acrosome-intact sperm (PMAI), sperm showing a high degree of DNA fragmentation (%DFI)) were performed after 0 and 3xa0h of incubation after thawing. After an equilibration time of 48xa0h and 72xa0h values for PMAI0h were higher (Pxa0<xa00.05) compared to PMAI0h values of sperm equilibrated for 6xa0h, and %DFI3h values were higher after 96xa0h (Pxa0<xa00.05) compared to 6xa0h equilibration. Between 12 and 90xa0TS and 13-97 CS were tightly bound to each HZ, respectively. The mean Hemizona Index (HZI) after a sperm equilibration for 48xa0h (HZIxa0=xa092.3xa0±xa012.7) or 72xa0h (HZIxa0=xa098.9xa0±xa016.23) was higher (Pxa0<xa00.01) than after an equilibration for 6xa0h (HZIxa0=xa073.3xa0±xa013.93) or 96xa0h (HZIxa0=xa081.3xa0±xa011.41). The HZI for 96xa0h equilibration was moderately negatively related to PMS0h and PMS3h (rxa0<xa0-0.35, Pxa0<xa00.05). Furthermore the HZI for 6xa0h equilibration was highly negatively correlated with DFI0h (rxa0=xa0-0,46, Pxa0<xa00.01). On the basis of these results it can be concluded that the hemi-zona assay is a suitable test to detect alterations in the fertilizing capacity of bovine sperm after modifying the equilibration period.

Correlation between endoscopic and histopathological findings in dogs with chronic gastritis

Abstract Introduction: Chronic gastritis is a common diagnosis in dogs with signs of chronic vomiting. However, there is no data concerning endoscopic and histopathological agreement in dogs with chronic gastritis. Thus, a question should be raised whether taking gastroduodenal biopsies in dogs with chronic gastritis is necessary or not. Consequently, the purpose of the study was to compare the endoscopic and histopathological agreement in dogs with chronic gastritis. Material and Methods: A total of 22 non-pregnant client-owned dogs with the signs of chronic gastritis were enrolled in this prospective study. Procedures including clinical examination, blood analysis, and diagnostic imaging were performed before anaesthesia. Biopsies obtained from gastroduodenal sites were histopathologically evaluated. A total of 110 gastroduodenal samples were examined. Results: Sixtyeight samples had abnormal histopathology and endoscopy while 11 showed normal histopathological and endoscopic evidence. Conclusion: The obtained data demonstrated that it is not necessary to take extra gastroduodenal biopsies in dogs with evidence of endoscopic gastroduodenitis. We also believe that further prospective studies, including cost and time effectiveness and more specific comparison between endoscopic appearance and histopathology, are necessary to make final recommendations regarding the need of using both procedures for definitive diagnosis.