Neuroprotective effects of dietary borax in the brain tissue of rainbow trout (Oncorhynchus mykiss) exposed to copper-induced toxicity.

We aimed to investigate the modulating effects of dietary borax on the pathways in rainbow trout brain exposed to copper. For this aim, a comprehensive assessment was performed including biochemical (acetylcholinesterase (AChE), malondialdehyde (MDA), oxidative DNA damage (8-hydroxy-2'-deoxyguanosine (8-OHdG), and caspase-3 levels) and transcriptional parameters (heat shock protein 70 (HSP70) and cytochromes P450 (CYP1A), glutathione peroxidase (gpx), superoxide dismutase (sod), and catalase (cat)) parameters and immunohistochemically staining of 8-OHdG. Special fish feed diets were prepared for the trial. These diets contained different concentrations of borax (1.25, 2.5, and 5 mg/kg) and/or copper (500 and 1000 mg/kg) at the period of pre- and co-treatment strategies for 21 days. At the end of the treatment periods, brain tissue was sampled for each experimental group. As a result, the biochemical parameters were increased and AChE activity decreased in the copper and copper-combined groups in comparison with the control group and also with only borax applications (p < 0.05). We observed an increase or decrease in particular biochemical parameters for the borax group in every application and we established that borax had protective effect against copper toxicity by decreasing and/or increasing the relevant biochemical parameters in brain tissue of fish. The biochemical results of borax and its combinations corresponded to the observations of gene expression data, which similarly concluded that HSP70 and CYP1A genes were strongly induced by copper (p < 0.05). In addition, the expression levels of the sod, cat, and gpx genes in the fish brains exposed to borax and the borax combination groups were significantly higher than the only copper-treated groups. In conclusion, borax supplementation provided significant protection against copper-induced neurotoxicity in trout.

Low transcriptional activity of PON2 in recurrent abortion: A novel therapeutic agent?

OBJECTIVE: Oxidative stress has been reported to be associated with various pregnancy complications and to play key roles in many of them. An inadequate level of antioxidant defense may eventually lead to an early pregnancy loss. There is a lack of information about the roles of the PON2 and PON3 enzymes in the etiology of the cases of unexplained recurrent abortus. The aim of our study is to determine and present the data regarding the roles of these enzymes for the first time. MATERIALS AND METHODS: We measured the transcriptional levels of the PON2 and PON3 enzymes in the curettage materials obtained from the patients with unexplained recurrent abortus (n=25) and compared the results with those measured in the abortus materials from healthy pregnant women (n=50) who had undergone a voluntary abortion. The transcriptional activities of PON2 and PON3 enzymes were measured through quantification of their respective mRNAs by RT-qPCR assay. For each gene, 2-ΔCt replication values of the control and the patient groups were compared using the Student's t-test, and the p values were calculated thereafter. Fold-changes in the enzyme transcription levels were interpreted as up- or down-regulation. RESULTS: PON2 mRNA expressions were found to be highly decreased in the patient group (p=0.000002). PON3 transcription, when compared to the healthy pregnant women, was found to be down-regulated in the patient group; however, the difference was not statistically significant (p=0.69). CONCLUSIONS: In this study, we evaluated the expressional regulation of the PON2 and PON3 enzymes in unexplained recurrent abortus. Our results demonstrate for the first time that the expressions of PON2 and PON3 are down-regulated in the abortion specimens of the patients with recurrent miscarriage. Although both enzymes had low expression levels, the decrease in the transcriptional activity of PON2 revealed a high statistical significance. According to these results, it is rational to speculate that PON2 may be a novel therapeutic agent in the management of the cases with unexplained recurrent abortion.

Assessment of 8-hydroxy-2-deoxyguanosine activity, gene expression and antioxidant enzyme activity on rainbow trout (Oncorhynchus mykiss) tissues exposed to biopesticide.

The goal of this study was to determinate toxicity mechanism of biopesticide with antioxidant enzymes parameters such as superoxide dismutase (SOD), glutathione peroxidase (GPx) and catalase (CAT) and malondialdehyde (MDA) levels, oxidative DNA damage (8-hydroxy-2-deoxyguanosine (8-OHdG)), transcriptional changes of heat shock protein 70 (HSP70), and cytochromes P4501A (CYP1A), sod, cat, and gpx in liver and gill tissues of Oncorhynchus mykiss. For this aim, plant-based (natural pesticides, azadirachtin (AZA)) and synthetic pesticides (deltamethrin (DLM)) were exposed on the fish at different concentrations (0.0005 and 0.00025ppm of DLM; 0.24 and 0.12ppm of AZA) for 21 days. According to the results of the study, the activity of SOD, CAT and GPx decreased, but malondialdehyde (MDA) level and activity of 8-OHdG increased in the gill and liver of rainbow trout (p<0.05). Additionally sod, cat and gpx were down regulated; HSP70 and CYP1A were up regulated for transcriptional observation. The downwards regulation of antioxidant (sod, cat and gpx) and the upregulation of HSP70 and CYP1A was obvious with doses of AZA or DLM (p<0.05). The findings of this study suggest that biopesticide can cause biochemical and physiological effects in the fish gill and liver by causing enzyme inhibition, an increase in 8-OHdG levels and changes in both transcriptional parameters (sod, cat, gpx, HSP70 and CYP1A). We found that excessive doses of plant-based pesticide are nearly as toxic as chemical ones for aquatic organisms. Moreover, 8-OHdG, HSP70 and CYP1A used as a biomarker to determinate toxicity mechanism of biopesticide in aquatic environment.

Investigation of 8-OHdG, CYP1A, HSP70 and transcriptional analyses of antioxidant defence system in liver tissues of rainbow trout exposed to eprinomectin.

Eprinomectin (EPM), a member of avermectin family, is a semi-synthetic antibiotic. It has been known that avermectin family enters the aquatic environments and adversely affects the aquatic organisms. Effects of EPM is fully unknown in aquatic organisms especially fish, thus the aim of the present study was to investigate transcriptional changes (sod, cat, gpx) and activities of some antioxidant enzymes (superoxide dismutase (SOD), glutathione peroxidase (GPx) and catalase (CAT) and malondialdehyde (MDA) levels, oxidative DNA damage (8-hydroxy-2-deoxyguanosine (8-OHdG)) and transcriptional changes of heat shock protein 70 (HSP70), and cytochromes P4501A (CYP1A) in liver tissues of rainbow trout exposed to sublethal EPM concentration (0.001 µg/L, 0.002 µg/L, 0.01 µg/L, 0.05 µg/L) for 24 h, 48 h, 72 h and 96 h. The decrease in antioxidant enzyme (SOD, CAT and GPx) activity, transcriptional changes (sod, cat, gpx, HSP70 and CYP1A genes) and increase in MDA level and activity of 8-OHdG in a dose-time-dependent manner in the liver of rainbow trout were observed. The down-regulated of antioxidant (sod, cat and gpx), HSP70 and CYP1A obviously, the severity of which increased with the concentration of EPM and exposure time. The results imply that EPM could induce oxidative damage to the liver tissue of rainbow trout. The information presented in this study is helpful to understand the mechanism of veterinary pharmaceuticals-induced oxidative stress in fishes.

Effect of Kineret® on ovarian ischemia reperfusion injury in a rat model.

AIM: To investigate the effect of Kineret® on ischemia reperfusion (IR) injury in rat ovaries. METHODS: Rats were divided into four groups: ovarian IR (IRG); 50 mg/kg Kineret® + ovarian IR (KIR-50); 100 mg/kg Kineret® + ovarian IR (KIR-100); and sham operation (SOC). KIR-50 (n = 10) and KIR-100 (n = 10) groups received an intraperitoneal injection of Kineret® at doses of 50 and 100 mg/kg, respectively. IRG and SOC (n = 10) rat groups were given distilled water as solvent using the same method. The results were compared between the groups. RESULTS: In rats in which IR occurred, oxidant parameters, such as malondialdehyde (MDA) and myeloperoxidase (MPO), were increased, the level of proinflammatory interleukin 1 beta (IL-1ß) was elevated and total glutathione (tGSH) as an antioxidant was decreased in the ovarian tissues. Administration of Kineret® at a dose of 100 mg/kg inhibited the increase of MDA, MOP and IL-1ß and a decrease in tGSH caused by IR more significantly than administration of Kineret® at a dose of 50 mg/kg. In addition, 100 mg/kg Kineret® significantly decreased severe hemorrhage, degeneration and inflammatory signs in the follicular cells, caused by IR. Kineret® at 100 mg/kg markedly ameliorated increased apoptosis in ovarian tissue with IR more significantly than 50 mg/kg kineret. CONCLUSION: Our findings indicate that Kineret® might be useful in clinical practice for the treatment of damage that may occur as a result of ovarian torsion.