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The influence of trehalose, taurine, cysteamine and hyaluronan on ram semen. Microscopic and oxidative stress parameters after freeze-thawing process
Theriogenology Cilt 67 ss. 1060-1067
Scopus Toplam 341 atıf DOI
There is a lack of information regarding lipid peroxidation and antioxidant capacity in cryopreserved ram semen, and cryopreservation is associated with the production of reactive oxygen species (ROS) which lead to lipid peroxidation (LPO) of sperm membranes, resulting in a loss of motility, viability and fertility of sperm. The aim of this study was to determine the influence of certain additives and their different doses on standard semen parameters, lipid peroxidation and antioxidant activities after the cryopreservation/thawing of ram semen. Ejaculates collected from four Akkaraman rams, a native breed of sheep, were evaluated and pooled at 33 °C. Semen samples which were diluted with a Tris-based extender containing additives including trehalose (50, 100 mM), taurine (25, 50 mM), cysteamine (5, 10 mM), and hyaluronan (0.5, 1 mg/ml), and an extender containing no additives (control) were cooled to 5 °C and frozen in 0.25 ml French straws, being stored in liquid nitrogen. Frozen straws were thawed individually at 37 °C for 20 s in a water bath for evaluation. The use of a Tris-based extender supplemented with 50 mM trehalose, 25 mM taurine, and 5 and 10 mM cysteamine led to higher percentages of post-thaw motility, in comparison to the control group (P < 0.01). No significant differences were observed in the percentages of acrosome and total abnormalities, and the hypoosmotic swelling test upon the supplementation of the freezing extender with antioxidants after the thawing of semen. In biochemical assays, the addition of antioxidants did not cause significant differences in levels of malondialdehyde (MDA), glutathione (GSH), and glutathione peroxidase (GSH-Px), after thawing, when compared to groups with no additives. In this study, catalase (CAT) activities were higher in the group that was applied 25 mM taurine as an antioxidant, than in all of the other groups (P < 0.001). Compared to the controls, antioxidant treatment with 100 mM trehalose, 50 mM taurine, 5 mM cysteamine and 0.5 mg/ml hyaluronan, significantly elevated vitamin E (vit E) levels in samples (P < 0.001).
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IMMUNOFLUORESCENCE AND BIOCHEMICAL INVESTIGATION OF THE PROTECTIVE EFFECTS OF NARINGIN AND DIOSMIN ON THE FREEZABILITY OF MERINO RAM SEMEN
Cryo Letters Cilt 45 ss. 309-319
Scopus Havuzumuzda 2 atıf almış
BACKGROUND: Various antioxidant substances are added to sperm extenders to protect spermatozoa against oxidative stress and cryodamage. OBJECTIVE: To investigate the effects of the flavonoid diosmin (DIO) and a flavanone glycoside naringin (NAR) on the freezability of ram semen. MATERIALS AND METHODS: In this study, six Merino rams were used during the breeding season. The ejaculates were pooled after collection from the rams. Pooled ejaculates were divided into six groups: control, NAR 1 mM, NAR 2 mM, NAR 4 mM, DIO 2 mM, and DIO 4 mM, and then diluted with a TRIS-based diluent. The pooled semen was equilibrated, placed in 0.25 mL pipettes with 10 × 107 sperm cells in each pipette, and frozen in liquid nitrogen vapor. After 24 h, the pipettes were thawed at 37°C for 25 s and analyzed in terms of spermatological parameters. RESULTS: The highest plasma membrane integrity ratio was found in the DIO 4 mM group, whereas a statistically significant difference was found between the NAR 1 mM and NAR 2 mM groups (p<0.05). While the DIO 4 mM group had the highest acrosome integrity rate, a statistically significant difference was found between the other groups (p<0.05). Mitochondrial activity was the highest in the NAR 4 mM, DIO 4 mM and DIO 2 mM groups (p<0.05). In the analysis of the sperm membrane lipid profile, it was observed that the DIO group had the highest lipid-phospholipid ratio. In sperm membrane protein profile analysis, it was found that both additives exerted protective effects at different levels. The highest total protein content was seen in the DIO 4 mM and NAR 4 mM groups. 8-hydroxydeoxyguanosine (8-OhDG) positivity was more common in the control group than in the DIO and NAR groups. Cu-Zn superoxide dismutase (SOD) expression was lower in the control group and more intense in all other groups. Positive results were especially observed in the acrosome of the sperm cells. CONCLUSION: The addition of NAR and DIO to the ram semen extender increased the quality of sperm parameters after the freeze-thaw process.
Atıf Yapan Makale Bilgileri
Kurumlar (6)
Atatürk Üniversitesi Erzurum, Turkey
Cumhuriyet Üniversitesi Sivas, Turkey
Erciyes Üniversitesi Kayseri, Turkey
Kastamonu University Kastamonu, Turkey
Provincial Directorate of Agriculture Mus, Turkey
Selçuk Üniversitesi Selçuklu, Turkey