Effect of cholesterol loaded cyclodextrin on semen cryopreservation of Aksaray Malakli shepherd dogs of different ages.

The objective of the study was to determine the effect of cholesterol-loaded cyclodextrin (CLC) on the quality parameters of semen from Aksaray Malakli Shepherd dogs of different age groups. Forty-eight male dogs were divided into 3 groupings according to their ages (young age (Y): ≤3 years, n: 20; middle age (M): 4-6 years, n: 20; old age (O): ≥7 years; n: 8). The sperm-rich portion of the ejaculate from each dog was divided into four aliquots and extended with either tris as a control (C) or tris loaded with 0.5, 1.0, and 1.5 mg/120 × 106 CLC as low (L), intermediate (I), and high (H) doses, respectively. Following equilibration for at least half an hour, the straws were frozen in nitrogen vapor and then stored in liquid nitrogen at least for 48 h. Later, the frozen straws were thawed in a water bath for spermatological evaluation. Significant differences were observed between different age groups in terms of the spermatological parameters (p < 0.05). The evidence suggests that increasing age is associated with poor in-vitro spermatological parameters and CLC was able to protect the acrosome integrity from cryo-damage during the freeze-thawing process. Better semen freezability characteristics were obtained at young ages, considering the overall parameters.

Metal pollution in biotic and abiotic samples of the Büyük Menderes River, Turkey.

The Büyük Menderes River (BMR) is one of the largest rivers in Turkey. This river irrigates efficient farmlands and includes tributaries of other rivers and streams and many populated towns within its limits in the Ege region. Both the estuary and Isikli Lake serve as a sanctuary for various waterbirds. Therefore, the BMR plays a critical role both for the inhabitants and for the ecosystem organisms in its environs. In the present study, we analyzed levels of metals including iron, barium, zinc, vanadium, cobalt, chromium, cadmium, copper, nickel, aluminum, arsenic, manganese, antimony, silver, selenium, boron, mercury, titanium, and lead in river water, sediment, fish (Cyprinus carpio; common carp), and in various waterbird (Fulica atra, Euroasian coot; Larus michahellis, yellow-legged gull; Ardea cinerea, grey heron; Larus melanocephalus, Mediterranean gull; and Pelecanus crispus, pelican) samples. Analyses were performed using an inductively coupled plasma-mass spectrometry (ICP-MS) instrument after sample preparation. Comparing metal concentrations among different sample types, it was found that barium, aluminum, and zinc are the major metals in river water, and zinc in common carp muscle, while iron, aluminum, and manganese are the major metals in sediments. Iron, zinc, copper, and aluminum were the highest in waterbird muscle tissue. Iron and barium were found to be the major metals in eggshell, while iron and zinc are the major metals in egg samples. A simple "worst-case scenario" model of risk assessment revealed that some of the analyzed metals may pose a risk for human health through consuming fish.

Comparing ethylene glycol with glycerol and with or without dithiothreitol and sucrose for cryopreservation of bull semen in egg-yolk containing extenders.

There are few studies performed for investigating the roles of different ratio and cryoprotectants with dithiothreitol or sucrose on sperm motility characteristics and antioxidant capacities of post-thawed bull spermatozoa. The objectives of this study were to compare glycerol (G) and ethylene glycol (EG) at different concentrations as cryoprotectants and dithiothreitol (D) or sucrose (S) (with/without) as antioxidants in Tris extender for cryopreservation of bull semen. Twenty-four ejaculates obtained from three bulls were included in the study. Each ejaculate was split into four equal aliquots and diluted using both of the Tris extenders with glycerol (5% or 7%) or ethylene glycol (3% or 5%). After that, each extenders were split into three equal aliquots and diluted using both of the dithiothreitol 5mM or sucrose 25 mM, and control (without additives) was cooled to 4 °C and frozen in 0.25-ml French straws. when compared to control, different doses cryoprotectants and antioxidants addition no significantly increased the percentages of post-thaw sperm progressive and motitilities, acrosome abnormality and plasma membrane integrity (P>0.05). However, EG3+S yielded the greatest percentages of the total abnormality (P<0.05). As regard to antioxidant activities G7 and EG5 led to lowest MDA activity with or without D or S but, these results were not supported to the GPx activity (P<0.01). The sperm motion characteristics such as VAP, VCL, ALH and BCF gave significantly different results (P<0.05). When compared the DNA integrity, different doses cryoprotectants without antioxidants addition significantly increased the percentages of the tail intensity and tail moment (P<0.05). There were no significant differences observed in non-return rates among all treatment groups (P>0.05).

Can linoleic acid improve the quality of frozen thawed bull sperm?

BACKGROUND: Cryopreservation is known to have a detrimental effect on the motility, viability and membrane integrity of sperm cells. OBJECTIVE: The aim of this study was to investigate the effect of various amount of linoleic acid supplementation to the Tris extender, on bull sperm parameters, DNA integrity and oxidative stress after freeze-thawing. METHODS: Ejaculates were split into five aliquots and extended to a final concentration of 18x10(6) spermatozoa per ml with the base extender containing different doses of linoleic acid 0.125 ml, (L125); 0.250 ml (L250); 0.5 ml (L500), 1 ml (L1000) and no additive (control; L0). The extended samples were equilibrated slowly to 4 degree C for 4 h and then froze using a digital freezing machine. Frozen straws were thawed individually in water bath at 37 degree C for 30 s to analyse progressive motility and sperm motion characteristics as well as membrane integrity. Biochemical assays were performed in a spectrophotometer using commercial kits. DNA damage was evaluated by Comet Assay. RESULT: The addition of various linoleic acid did not improve the sperm subjective, CASA and progressive motilities, sperm motility characteristics and DNA integrity (P>0.05). L500 exhibited the greatest values for membrane integrity than that of the other groups (P<0.001). All supplementation groups led to lower percentages of tail abnormalities in comparison to the control (P<0.001). L500 and L1000 significantly decreased total abnormalities. In conclusion, our findings showed that L500 linoleic acid supplementation in semen extender was of great beneficial effect on frozen-thawed bull semen in terms of morphology and plasma membrane integrity.

Effects of smoking during pregnancy on DNA damage and ROS level consequences in maternal and newborns' blood.

Some of the genotoxic/carcinogenic substances or metabolites in cigarette smoke are capable of passing through the placenta and harming a newborn's health. Smoking is also known as a factor in the formation of oxidative damage and the main mechanism involved in the carcinogenic process. Predetermining this genotoxic risk can be successfully achieved by measuring certain parameters of oxidative stress. The comet assay is considered an important biomarker for the evaluation of genotoxic substances and is effective for detecting DNA damage caused by smoking. This study examined third trimester bloods and the cord blood of 28 actively smoking and 22 non-smoking mothers in terms of DNA damage and oxidative stress parameters. Cu/Zn superoxide dismutase (CuZn-SOD), malondialdehyde (MDA), catalase (CAT), plasma nitrite/nitrates (NO2-/NO3-), selenium-dependent glutathione peroxidase (Se-GPx), Cu, and Zn levels were measured as indicators of oxidative damage. There were no significant increases in DNA damage of the actively smoking pregnant group in comparison with the non-smoking pregnant group, either in the third trimester or cord blood. Oxidative stress parameters of smoker and non-smoker groups were statistically different for MDA (p<0.05), CuZn-SOD (p<0.01), Se-GPx (p<0.05) values while the difference was not significant for NO2-/NO3-, CAT, Zn, and Cu values. The same values were also investigated in cord blood,and only NO2/NO3-(p<0.01), Se-GPx (p<0.01 and CAT (p<0.001) values were found statistically different.Smoking mothers may have been exposed to more oxidative stress than non-smoking mothers

Assessment of individual susceptibility to baseline DNA and cytogenetic damage in a healthy Turkish population: evaluation with lifestyle factors.

BACKGROUND: Cytogenetic biomarkers are most frequently used well-established endpoints in human population studies with their sensitivity for measuring exposure to genotoxic agents. They have an important role as early predictors of cancer risk. Identification of individual genotypes of metabolic gene polymorphisms helps to understand the modulation of cancer susceptibility by environmental exposures, such as cigarette smoking and other lifestyle factors. AIM: To evaluate individual susceptibility to chemicals, we determined individual DNA damage related to glutathione S-transferase (GST) genotypes (GSTM1, GSTT1, and GSTP1) in a Turkish population. METHODS: Peripheral blood lymphocytes (PBL) and DNA samples of 127 subjects were analyzed for the presence of DNA damage, using single-cell gel electrophoresis (the Comet assay), and for cytogenetic parameters (chromosomal aberrations [CAs], bleomycin-induced CA, and a cytokinesis-blocked micronucleus assay), and the polymerase chain reaction/restriction fragment length polymorphism method, respectively. RESULTS: Individuals carrying a GSTT1-null allele showed higher frequencies of CA and micronucleus (MN) (p=0.026, p=0.003, respectively), whereas the GSTM1-null and GSTP1 mutant genotypes did not show any differences in cytogenetic parameters. Our findings demonstrated that none of the lifestyle factors (smoking, alcohol drinking, dietary habits, vitamin intake, and physical activity), except for vitamin intake (p=0.002), were significantly associated with the studied cytogenetic parameters. CONCLUSION: Our results suggest that the GSTT1 gene polymorphism may influence the baseline cytogenetic frequency in a healthy population.

Evaluation of DNA damage in common carp (Cyprinus carpio L.) by comet assay for determination of possible pollution in Lake Mogan (Ankara).

Contamination of the aquatic environment with various concentrations of pollutants results in unexpected threats to humans and wildlife. The consequences of exposure and metabolism of pollutants/xenobiotics, especially carcinogens and mutagens, can be suitably assessed by investigating severe events, such as DNA damage; for example, DNA adducts and DNA strand breaks. One of the commonly used techniques to detect DNA damage in aquatic organisms is single-cell gel electrophoresis (comet assay). This study was carried out using Cyprinus carpio in order to identify the possible pollution in Lake Mogan, near Ankara, Turkey, where the city's sewer system and pesticides used in agriculture are believed to be the common causes of pollution. From the comet assay, the tail length (micrometer), tail intensity (%), and tail moment values of fish caught from Lake Mogan were found to be 31.10 ± 10.39, 7.77 ± 4.51, 1.50 ± 1.48, respectively, whereas for clean reference sites they were found to be 22.80 ± 1.08, 3.47 ± 1.59, 0.40 ± 0.51, respectively. The values are statistically different from each other (p < 0.0001, p < 0.0001, and p = 0.0013, respectively). These results indicate that Lake Mogan may be polluted with substances that have genotoxic effects and constitute an early warning for the lake system. Further detailed research is needed to establish the source of the pollution and the chemicals responsible.

Polychlorinated biphenyl and organochlorine pesticide levels in human breast milk from the Mediterranean city Antalya, Turkey.

In this study, organochlorine pesticides (OCPs) and polychlorinated biphenyls (PCBs) levels were determined in 100 human milk samples from the city of Antalya. The levels of seven major PCB congeners; 28, 52, 101, 118, 138, 153, 180 and nine OCPs, α-HCH, ß-HCH, γ-HCH, HCB, heptachlor epoxide, p.p'-DDT, p,p'-DDE, endosulfan-α and endosulfan-ß were determined by gas chromatography with ECD detection. The levels of analyzed compounds were as follows: ΣPCBs 27.46 ± 11.58, ΣDDT 1,407 ± 123, and ΣBHC 160 ± 490 ng/g lipid wt.basis. PCB 153 and p,p'-DDE were the dominant contaminants. The results have been discussed and compared with similar studies from other regions of Turkey.

An in vivo assessment of the genotoxic potential of bisphenol A and 4-tert-octylphenol in rats.

Bisphenol A (BPA) and octylphenol (OP) are industrial chemicals used in the manufacture of polycarbonate plastics, epoxy resins, and non-ionic surfactants. In the present study, we investigated the possible in vivo genotoxic effects of these compounds in rats using single-cell gel electrophoresis, the so-called comet assay. Adult male Wistar albino rats were divided randomly into six groups as follows: BPA125 (received 125 mg/kg bw BPA; n = 6), OP125 (received 125 mg/kg bw OP; n = 6), BPA250 (received 250 mg/kg BPA; n = 6), OP250 (received 250 mg/kg bw OP; n = 6), control (n = 5), and MMS (positive control group that received methyl methanesulfonate; n = 3). Both BPA and OP were orally administrated for 4 weeks. Controls were orally inoculated with corn oil for 4 weeks as well. Comet parameters including tail length and tail moment were evaluated for possible genotoxic effects. There were no significant differences in the OP125 and in the BPA125 compared with the control group, regarding tail length and tail moment (P > 0.05). However, there were significant differences in the OP250 and in the BPA250 compared with the control group, regarding tail length and tail moment (P < 0.05 and P < 0.01, respectively). The genotoxic potential of BPA and OP was investigated in vivo; there is a need for further studies exploring further mechanisms of the genotoxic potential of these chemicals in vivo.

Genotoxicity of anti-tumor necrosis factor therapy in patients with juvenile idiopathic arthritis.

OBJECTIVE: To assess the possible effects of both inflammation and the anti-tumor necrosis factor agents (anti-TNF) on DNA damage with a specific assay, and their effects on the repair capacity of DNA. METHODS: From a group of 20 children with juvenile idiopathic arthritis (JIA), 16 patients who completed the study and 16 control subjects were evaluated. DNA damage and repair capacity were analyzed by the comet assay at the level of peripheral lymphocytes before anti-TNF (etanercept) injections and on the 15th, 90th, and 180th days after the first injection. RESULTS: The amount of damage as detected by the aforementioned assay was higher in patients with JIA compared with controls. On the 15th day after the initial anti-TNF injection, there was a decrease in the mean DNA tail length of JIA patients, however on the 90th day an increase was observed; thereafter, an upward trend was observed until the end of the study. JIA patients had a DNA repair capacity that was significantly less than that of controls. CONCLUSION: The results of the comet technique suggests that JIA patients already have increased basal DNA damage before anti-TNF therapy; they are more sensitive to the DNA damage produced by H(2)O(2), and have a less efficient DNA repair system in comparison with control cells. After an initial improvement at 2 weeks, parameters of genotoxicity worsened, and DNA repair was further impaired 6 months after the addition of an anti-TNF agent to treatment.

Determination of organochlorine pesticide and polychlorinated biphenyl levels in adipose tissue of infertile men.

During the recent two decades, a remarkable drop in fertility rates has been noticed almost all over the world. A series of studies have showed that environmental factors had the primary role causing the observed adverse trends in the male reproductive health problems. Especially chlorinated hydrocarbons, for instance polychlorinated biphenyls (PCB) and pesticides, could mimic or antagonize the effects of steroid hormones, like estrogens and androgens and possibly interfering with male reproductive capacity. Organochlorine pesticides (OCPs) (i.e. p.p'-DDT, endosulfan...) and PCBs are widespread used chemicals in agriculture and industry for different purposes all over the world. Although direct evidence is lacking, theoretical considerations and epidemiological evidence implicate these compounds as potential hazards to human and wildlife reproductive health. For this purpose, in this study adipose tissue samples have provided from healthy males which were diagnosed as infertile men living in Ankara, Turkey at least for 5 years. Residual levels of OCPs (alpha-BHC, beta-BHC, gamma-BHC, HCB, Endosulfan I, II, p,p'-DDE, and p,p'-DDT) and seven major persistent PCB congeners (PCB 28, 52, 101, 118, 138, 153, and 180) were measured in 25 infertile men and 21 healthy men's adipose tissue samples by GC-ECD. Levels of OCPs and PCBs in adipose tissue of infertile men compared with those provided from controls. When the concentrations of each of the OCP were compared between the fertile and infertile groups, no statistical significance was obtained. Concentrations of each of the PCB congeners were compared between the fertile and infertile groups, no statistical significance was obtained (p > 0.05), except for 2,2',5,5'-Tetrachlorobiphenyl (PCB 52) (p = 0.032) and 2,2',3,4,4',5,5'-heptachlorobiphenyl (PCB 180) (p = 0.017).

beta2 adrenoceptor (ADRB2) pharmacogenetics and cardiovascular phenotypes during laryngoscopy and tracheal intubation.

Pharmacogenetics is the study of genetic variations that cause a variable drug response characterized by alteration in drug metabolism or in pharmacodynamics. The polymorphisms in genes encoding receptors relevant to treatment cause variation in sensitivity to many drugs. beta2 Adrenoceptor genetic variation contributes to regulation of blood pressure and hemodynamic changes by mediating peripheral vasodilatation. Laryngoscopy and tracheal intubation associated with hemodynamic changes. Although there are four nonsynomic single-nucleotide polymorphisms (SNPs) of beta2 adrenoceptor gene, codon 16 (Arg16Gly) and codon 27 (Gln27Glu) SNPs are both common and functionally important. In this paper, the authors investigated the beta2 adrenoceptor Gly16 and Glu27 SNPs in response to drugs relevant to anesthesia and how these SNPs impacted upon the cardiovascular phenotypes. The authors measured arterial systolic and diastolic blood pressure, heart rate, and rate-pressure product before induction of anesthesia and 1 min following laryngoscopy and tracheal intubation. Genomic DNA was amplified and genotyped using allele-specific polymerase chain reaction (ASPCR) and restriction fragment length polymorphism (RFLP) assays, respectively. When the authors compared hemodynamic results according to genotypes, the patients with Gln homozygote allele at codon 27 exhibited significant increase of heart rate than patients with Glu allele after laryngoscope and tracheal intubation.