PRNP gene polymorphisms in main indigenous Turkish goat breeds.

The polymorphisms of the PRNP gene influence the susceptibility to scrapie in goats. In this study, caprine PRNP gene was analysed in a total of 249 individuals from three main indigenous goat breeds of Turkey: Anatolian Black, Angora and Kilis. We focused on the Anatolian Black breed, which represents 97% of the goat population in Turkey and compared the data of samples originated from different geographical regions. Eight polymorphisms were determined, given rise to 12 haplotypes. Allele, genotype and haplotype frequencies of the polymorphisms at codons 142, 143, 146, 154, 171, 211, 222 and 240 were calculated. Alleles associated to resistance to scrapie were found to be relatively rare in all breeds. The resistance allele 222K was absent in Turkish breeds. Other resistance-associated alleles: 146D, 146S, 154H and 171R were observed with low frequencies. The results of this study, which cover the mainly bred indigenous goats in Turkey, present the distribution of PRNP polymorphisms. Very low frequencies of resistance-associated alleles show the susceptibility to scrapie. The resistance-associated alleles S and D of codon 146 might be accepted as candidate alleles, due to their relative higher frequencies observed in the present study. A breeding program aiming to increase particularly the frequency of 146S might be applied. Predictions about impacts of a long-term breeding programme based on low initial allele frequencies and regarding its possible adverse effects are warranted. Our results might be a database for future breeding programmes, which should be carefully designed with adequate levels of genetic resistance and acceptable timeframe.

Effects of isoxsuprine hydrochloride on electrocardiographic and trace element status in sheeps.

OBJECTIVE: To study the effect of isoxsuprine hydrochloride on the ischaemic electrocardiographic change and trace element status in sheep. METHODS: This study was conducted from March 16 to 23, 2012, at Istanbul University, Turkey, and comprised sheep aged 6 months. The animals were divided into two equal groups. The control group was fed a standard diet and had free access to water. In the experimental group, isoxsuprine hydrochloride was injected at a dose of 0.6 mg/kg through the intramuscular route. Electrocardiographic changes, including creatine kinase and cardiac troponin-I, and serum levels of selenium, copper, calcium, magnesium, iron and zinc were investigated in healthy sheep. SPSS 15 was used for statistical analysis. RESULTS: The 14 sheep were divided into two groups of 7(50%) each. The overall mean weight of the study population was 35±10kg. Selenium, calcium, iron and zinc concentrations did not show any difference in serum samples (p>0.05). However, copper and magnesium concentrations decreased in serum after the administration of the drug (p<0.05). In the experimental group, ST segment depression and abnormal T-wave was found in 6(86%) animals within 60min. CONCLUSIONS: Isoxsuprine hydrochloride increased cardiotoxicity risk in sheep.

In vitro effects of doxorubicin and deracoxib on oxidative-stress-related parameters in canine mammary carcinoma cells.

The present study evaluated the effects of doxorubicin (DOX) and deracoxib (DER), as single agents and in combination treatments, on antioxidant parameters in the canine mammary carcinoma cell line CMT-U27. The cells were exposed to DOX and DER for 24, 48 and 72 h. The viability and malondialdehyde (MDA), nitric oxide (NO), catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GSHPx) and total glutathione (GSH) activities of CMT-U27 cells were determined. The half inhibition concentration (IC50) of DOX was found to be ∼0.9 µM in the 72-h period. IC50 and 1/10 IC50 concentrations of DOX were combined with all concentrations of DER (50-1000 µM) in the combination experiments. The results showed increased oxidative status associated with significant decreases of CAT and GSH levels in CMT-U27 cells exposed to 10-µM and higher concentrations of DOX compared to control cells. In contrast, there were no significant changes in the groups tested with any of the concentrations of DER (50-1000 µM). In combination treatments, DER attenuated DOX-induced oxidative damage by modulating the enzymatic and non-enzymatic components in CMT-U27 cells. We suggest that the combination of DOX and DER can be beneficial in the treatment of cancer cells by increasing cellular responses to oxidative stress. In conclusion, the use of COX inhibitor in conjunction with a chemotherapeutic agent may provide a basis for new concepts of cancer treatment through systematic modulation of the antioxidant defence systems in mammary cancers of animals.

FASN gene polymorphism in indigenous cattle breeds of Turkey.

The aim of this study was to determine the variants of the thioesterase (TE) beta-ketoacyl reductase (KR) domains of the Fatty Acid Synthase (FASN) gene, in the East Anatolian Red (EAR) and South Anatolian Red (SAR) cattle breeds. It has been suggested that the FASN gene is effective on fatty acid composition of meat in cattle. In this study, the genotype and allele frequencies of g.17924 A>G, g.18440 G>A and g.16024 G>A, g.16039 T>C in TE and KR domains, respectively, were detected by using polymerase chain reaction - restriction fragment length polymorphism (PCR-RFLP) method. The g.18663 T>C polymorphism of the TE domain was determined by direct sequencing. The GG genotype of the g.17924 A>G polymorphism, which affects unsaturated fatty acid composition positively, has a high frequency in EAR and SAR breeds. The frequencies of the two haplotypes g.16024 G>A and g.16039 T>C in the KR domain were found to be significantly high in both breeds. These haplotypes also have positive effects on unsaturated fatty acid composition. The AA genotype of the g. 18440 G>A polymorphism, which is suggested to be absent in Bos taurus breeds, was detected in SAR and EAR breeds with frequencies close to those in Bos indicus breeds. In conclusion, we suggest that SAR and EAR cattle breeds have an advantage in terms of genotype and haplotype distribution of the polymorphisms in TE and KR domains of the FASN gene. Additionally g.18440 G>A polymorphism might be a potential marker for breed discrimination.

In vivo assessment of antidiabetic and antioxidant activities of rosemary (Rosmarinus officinalis) in alloxan-diabetic rabbits.

Rosemary (Rosmarinus officinalis), used in traditional Turkish folk medicine for the treatment of hyperglycaemia, is widely accepted as one of the medicinal herb with the highest antioxidant activity. Accordingly, the present study was designed to investigate the possible actions of ethanolic extract of the leaves of Rosmarinus officinalis on glucose homeostasis and antioxidant defense in rabbits. In the first set of experiments, hypoglycaemic effects of oral administration of various doses (50, 100 and 200 mg/kg) of the extract were examined in normoglycaemic and glucose-hyperglycaemic rabbits. Optimal effect was observed in both of the animal groups with a dose of 200 mg/kg of the extract and this activity was independent from the effects of insulin. In another part of experiments, acute effect of various doses of the Rosmarinus officinalis extract on blood glucose and serum insulin levels was studied in alloxan-induced diabetic rabbits. Of the three doses of extract, the highest dose (200 mg/kg) significantly lowered blood glucose level and increased serum insulin concentration in alloxan-diabetic rabbits. The last set of experiments designed to investigate the subacute effect of the Rosmarinus officinalis extract on repeated administration in alloxan-diabetic rabbits. At the doses of 100 and 200 mg/kg, antihyperglycaemic effect of extract was accompanied by a significant increase in serum insulin levels in diabetic rabbits. Furthermore, during 1 week of treatment of diabetic rabbits with a dose of 200 mg/kg of the extract showed that the extract possessed a capability to inhibit the lipid peroxidation and activate the antioxidant enzymes. It was concluded that probably, due to its potent antioxidant properties, the Rosmarinus officinalis extract exerts remarkable antidiabetogenic effect.

Tumour preventive effect of Aloe vera leaf pulp lectin (Aloctin I) on Ehrlich ascites tumours in mice.

In this study, the prophylactic effect of the main lectin present in Aloe vera leaf pulp extract (Aloctin I) was assayed against Ehrlich ascites tumours in mice. The lectin administered prophylactically before tumour implantation regressed tumour size, however, this activity was less potent than that of the A. vera leaf pulp extract previously shown in our laboratory. Accordingly, serum sialic acid and tumour necrosis factor alpha (TNFalpha) levels, chosen as tumour markers, were decreased significantly by the prophylactic administration of the lectin. The increase in spleen and thymus weights in the group given only Aloctin I, could be explained by the immunomodulatory and mitogenic effects of lectins. These findings, along with lymphoid hyperplasia observed in spleen and thymus, suggest that the tumour preventive effect of Aloctin I could be due to its immunomodulatory activity.

Effect of Aloe vera leaf pulp extract on Ehrlich ascites tumours in mice.

Among the various known therapeutic effects of Aloe vera (L.) Burm. fil., a few recent studies have shown that preparations of the plant leaves can prevent or regress the growth of certain tumours. In this study, undertaken with A. vera leaf pulp extract against Ehrlich ascites tumours in mice, the animals were separated into five groups: I - healthy control, II - tumour control, III - experiment 1 (extract given before tumour inoculation), IV - experiment 2 (extract given with tumour inoculation) and V - experiment 3 (extract given after tumour inoculation). Ehrlich ascites tumours (0.33 ml) were injected subcutaneously into groups II-V. Aloe extract was injected at 55 mg protein/kg, twice a week for 21 days. Tumour size, thymus and spleen weights were measured, as well as leucocyte count, tumour necrosis factor-alpha and sialic acid as tumour markers. The best inhibitory effect on tumour growth was obtained with the extract given prophylactically before tumour implantation (experiment 1), although Aloe extract also regressed tumour sizes when given simultaneously with (experiment 2), or therapeutically after (experiment 3), tumour implantation. Accordingly, serum sialic acid and tumour necrosis factor-alpha levels, chosen as tumour markers, which were raised in the tumour control group, were significantly decreased by the prophylactic administration of the extract. The increase in leucocyte count seen in experiment 1 and 2 groups, along with lymphoid hyperplasia observed in spleen and thymus necroscopy, lead us to think that the tumour preventive effect of Aloe could be due to its immunomodulatory activity. According to our results, A. vera could be proposed as a prophylactic for cancer prevention.