The cryoprotective potential of propolis supplemented in frozen-thawed bull semen; biochemical and physiological findings.

In this study, the cryoprotective effect of different doses of propolis (P) on bull semen, which has solid pharmacological properties thanks to its rich phenolic components, was investigated biochemically and physiologically. Semen samples were collected from Simmental breed bulls via the artificial vagina and pooled. After dividing into five groups, control (C: no additive) and four different P (200, 100, 50, and 25 µg/mL) groups, the final concentration was diluted to 16×106 per straw. Semen samples were equilibrated at 4°C for approximately 4 hours, then placed in French straws and frozen. After thawing, sperm motility and kinetic parameters, DNA integrity by single-cell gel electrophoresis, sperm abnormalities by liquid fixation, and lipid peroxidation levels by the colorimetric method was analyzed by Computer-Assisted Semen Analyzer. P added to the diluent showed no effect on motility and kinetic parameters at P25 and P50 (p>0.05), while P100 and P200 had a negative effect (p⟨0.001). The addition of P (25 and 50) showed a treatment effect on tail abnormality compared to C (p⟨0.05). Especially P50 had a positive effect on tail length, tail DNA, and tail movement, while P100 and P200 caused DNA damage (p⟨0.001). MDA levels increased in all P dose groups compared to C (p⟨0.001). This study has clearly demonstrated that P25 and P50 supplements could be used therapeutically to treat sperm tail abnormalities and prevent DNA damage in post-thawed bull sperm.

Supplementation of Rosmarinic Acid Has Reduced Oxidative Stress on Bull Spermatozoa Following the Freeze Thawing Process.

BACKGROUND: Cryopreservation has a side effect on the motility, chromatin integrity and viability of sperm cells. OBJECTIVE: The present study investigated the effects of supplementation with rosmarinic acid (RA) Tris extender on sperm quality parameters, plasma and acrosome membrane damage, antioxidant enzyme activity and chromatin integrity following the freeze thawing process on bull spermatozoa. MATERIALS AND METHODS: Ejaculates were split into five aliquots and diluted to a final concentration of 15x106 spermatozoa per ml with the Tris extender containing RA (25, 50, 100 and 200 microgram per ml) and (control) and then frozen at a controlled rate. RESULT: Treatments did not give better results on the percentages of sperm progressive, total motility and sperm motion characters (P >0.05); however, RA25 and RA50 exhibited favourable chromatin integrity. In conclusion, RA25 and RA50 increased total antioxidant activity. As a consequence, the amount of MDA and chromatin damage were reduced in sperm cells.

Supplementation of quercetin for advanced DNA integrity in bull semen cryopreservation.

The aim of this study was to identify the effects of adding quercetin (Q) to Tris extender in order to identify levels of oxidative stress in bull sperm after freeze thawing. Ejaculates were collected via artificial vagina from Holstein bulls. Semen was divided into five tools and diluted to a final concentration of 15 × 106 spermatozoa/ml with the Tris extender containing Q (25, 50, 100 and 200 µg/ml) and no additive (control; C). All examples were equilibrated at 4°C during 4 hr then were loaded into 0.25-ml straws and frozen using a controlled rate. Sperm motility and motility characteristics were determined using the computer-assisted semen analyser. Sperm membrane integrity was assessed using the hypoosmotic swelling test. Sperm chromatin integrity was investigated using the single cell gel electrophoresis. Total antioxidant capacities were performed colorimetrically. Q supplementation used as an antioxidant did not produce better results in the proportion of sperm progressive and total motility, plasma membrane integrity and sperm abnormalities. Q supplementation exhibited the favourable tail length, tail DNA and tail moment. In conclusion, when whole parameters are considered, Q25 can be added to the Tris extender due to its positive effect on sperm DNA integrity and no adverse effect on the progressive and total motilities of sperm.

The clinical significance of endometrial fluid collection in asymptomatic postmenopausal women.

OBJECTIVE: This study assessed the clinical significance of endometrial fluid collection (EFC) in asymptomatic postmenopausal women. MATERIALS AND METHODS: A total of 564 postmenopausal females were enrolled into this retrospective study, of whom 141, with EFC, comprised the study group; the remaining 423 postmenopausal women (the three consecutive patients) who had been admitted to the menopause outpatient clinic for their routine annual check formed the control group. Data, including age, gravidity, parity, body mass index, hypertension, diabetes mellitus, menopause duration, endometrial thickness and histopathological endometrium results, were compared between groups using the SPSS for Windows software package (version 15.0; SPSS Inc., Chicago, IL, USA). RESULTS: There were no significant group differences in gravidity, parity, body mass index, rates of hypertension and diabetes mellitus, endometrial thickness or histopathological endometrium results. However, there were group differences in age (62.0 ± 7.6 vs. 57.2 ± 8.9 years, p < 0.001) and menopause duration (13.4 ± 7.8 vs. 7.9 ± 5.4 years, p < 0.001). CONCLUSION: The results indicate that EFC in asymptomatic postmenopausal women is a benign condition, influenced by older age and commensurately longer menopause duration due to a lack of hormonal stimulation. The presence of EFC by itself does not affect the decision to provide further evaluation; that is determined by the presence of confirmed risk factors (e.g. thickened endometrial stripe). However, further studies are needed to assess any long-term prognostic impact of this finding.

Comparison of cryoprotective effects of lycopene and cysteamine in different cryoprotectants on bull semen and fertility results.

The objectives of this study were to compare glycerol and ethylene glycol at different concentrations as cryoprotectants and lycopene or cysteamine (with/without) as antioxidants in Tris extender for bull semen. Twenty-four ejaculates were obtained from three bulls. 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 added using both of the cysteamine 5 mm or lycopene 500 µg/ml, and control (without additives). The addition of 7% glycerol with cysteamine, 5% ethylene glycol with cysteamine and 3% ethylene glycol with cysteamine groups gave the lowest CASA motility than the other groups. However, 7% glycerol and 7% glycerol with lycopene resulted in a better rate of CASA progressive motility compared with that of other groups. Generally, all the lycopene groups signed better protective effects on acrosome and total morphology than the other groups. Glycerol 7% and 3% ethylene glycol with lycopene groups yielded to slight higher percentages of membrane integrity assessed by HOST than that of the other groups, but 7% glycerol with cysteamine and 3% ethylene glycol with cysteamine showed the worst percentages of membrane integrity. Glycerol 7% and 5% glycerol with lycopene gave rise to a higher value of VAP, VSL and VCL compared with that of the other groups. On the contrary, adding to 5% glycerol with cysteamine showed negative effect for VAP, VSL, VCL and ALH values. All cryoprotectant groups with lycopene decreased chromatin damage than the other groups. Ethylene glycol 3% led to lower non-return rates of inseminated cows. However, this result was not considered to be statistically important.

Fertility in dairy cows after artificial insemination using sex-sorted sperm or conventional semen.

The aim of this study was to compare pregnancy per artificial insemination (P/AI) after timed AI with sex-sorted sperm (SS) or conventional semen (CS) in lactating dairy cows. Cyclic cows (n = 302) were synchronized by Ovsynch and randomly assigned into two groups at the time of AI. Cows with a follicle size between 12 and 18 mm and clear vaginal discharge at the time of AI were inseminated with either frozen-thawed SS (n = 148) or CS (n = 154) of the same bull. A shallow uterine insemination was performed into the uterine horn ipsilateral to the side of probable impending ovulation. Pregnancy per AI on Day 31 tended (p = 0.09) to be less for SS (31.8%) than CS (40.9%). Similarly, P/AI on Day 62 was less (p = 0.01) for cows inseminated with SS (25.7%) compared with CS (39.0%). The increased difference in fertility between treatments from Days 31 to 62 was caused by the greater (p = 0.02) pregnancy loss for cows receiving SS (19.2%) than CS (4.8%). Cow parity (p = 0.02) and season (p < 0.01) when AI was performed were additional factors affecting fertility. Primiparous cows had greater P/AI than multiparous cows both on Day 31 (41.7% vs 25.0% in SS and 53.0% vs 31.8% in CS groups) and on Day 62 (33.3% vs 20.5% in SS and 48.5% vs 31.8% in CS groups). During the hot season of the year, P/AI on Day 31 was reduced (p = 0.01) in the SS group (19.6%) when compared with the rates during the cool season (38.1%). In conclusion, sex-sorted sperm produced lower fertility results compared to conventional semen even after using some selection criteria to select most fertile cows.

Effects of different extenders and centrifugation/washing on postthaw microscopic-oxidative stress parameters and fertilizing ability of Angora buck sperm.

The main objective of the current study was to evaluate the effects of extender type and centrifugation/washing prior to cryopreservation on the postthaw sperm parameters, lipid peroxidation, and superoxide dismutase activity of Angora buck (Capra hircus ancryrensis) sperm. Ejaculates collected from three Angora bucks were used in this study. Two consecutive ejaculates from each buck were pooled and split into equal parts in four Falcon tubes. Two tubes were diluted at 37 degrees C and then centrifuged to remove semen plasma. After centrifugation, two sediment parts were diluted with a Tris-based extender and commercial Bioxcell extender, respectively. The remaining two parts, which were not centrifuged/washed, were diluted with the above-mentioned extenders, respectively. Diluted samples were cooled to 5 degrees C and frozen in 0.25-mL French straws to be stored in liquid nitrogen. Frozen straws were thawed individually at 37 degrees C for 20 sec in a water bath for evaluation. The semen part with centrifugation/washing in the Bioxcell extender (BC) demonstrated a higher rate of subjective motility (58.1+/-3.0%) compared with that of groups with (TC) or without (T) centrifugation/washing in the Tris-based extender (P<0.01). Angora buck sperm frozen with (BC) or without (B) centrifugation/washing in the Bioxcell extender demonstrated higher percentages of motility (60.6+/-2.7% and 54.3+/-4.8%, respectively) compared with that of groups T and TC. The postthaw progressive motility rate (22.3+/-2.7%) was significantly greater for semen parts diluted in B compared with that of other groups. BC gave rise to a lower value of average path velocity (90.0+/-5.2 microm/sec) compared with that of other groups (P<0.01). For straight linear velocity and linearity index, the highest values (103.2+/-4.7 microm/sec, 47.5+/-1.6% and 94.8+/-3.0 microm/sec, 44.8+/-1.1%, respectively) were obtained from B and TC (P<0.001). For sperm acrosome and total abnormalities, TC gave the highest values (11.2+/-0.6% and 26.6+/-1.5%, respectively, P<0.01). In the group frozen in BC, the percentage of membrane integrity assessed by hypo-osmotic swelling test was higher (61.2+/-2.2%) than that of the other groups (P<0.001). With respect to fertility results based on 35-d pregnancy rates, BC gave a higher rate (76.5%) than that of TC (27.8%, P<0.05). Malondialdehyde formation was found to be lower (1.64+/-0.26 nmol/L) in BC than in the other groups after the freeze-thawing process (P<0.001). In the semen part frozen in BC, superoxide dismutase activity was higher (0.18+/-0.02 U/mg protein) compared with that of the other groups (P<0.05). Further studies are required to obtain more precise results for the characterization of oxidative stress parameters and fertilizing ability in cryopreserved buck spermatozoa.

Prevalence and factors affecting the formation of second molar distal caries in a Turkish population.

The aim of this study was to evaluate the prevalence of second molar distal caries in a Turkish population and to determine the factors that affect it. Clinical records and panoramic radiographs of partially erupted mandibular third molars were reviewed in this retrospective study. The analysis outcome measures were the patients' age, second molar distal caries, third molar angulation and second and third molar contact point localization. Prevalence of second molar distal caries in the population was 20%. This prevalence was 47% when the third molar had an angulation of 31-70 degrees (majority of mesioangular third molars) and 43% at 70-90 degrees (all horizontal third molars). The contact point on the second molar amelocemental junction and increasing age had significant effects on caries formation. The results revealed that second molar distal caries justifies prophylactic third molar removal and partially erupted third molars that have an angulation of 30-90 degrees with a contact point on the amelocemental junction should be removed to prevent second molar distal caries.