High total antioxidant capacity of the porcine seminal plasma (SP-TAC) relates to sperm survival and fertility.
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ABSTRACT:
SUBMITTER: Barranco I
PROVIDER: S-EPMC4685244 | biostudies-other | 2015 Dec
REPOSITORIES: biostudies-other
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Project description:The escalated use of artificial insemination in swine has increased the importance of determining fertility of a semen sample before it is used. Multiple laboratory assays have been developed to assess fertilizing potential but they have yielded inconsistent results. This experiment sought to determine the relationship between in vitro competitive zona binding ability and in vivo fertility based on heterospermic inseminations and paternity testing. The zona pellucida binding ability and fertility of sperm from 15 boars was assessed by comparing sperm from one boar with sperm from other individual boars in a pairwise fashion using four ejaculates. The relationship of zona binding ability to the mean number of piglets sired per litter for each boar as well as historic fertility data (litter size and farrowing rate) was assessed. The in vitro competition assay consisted of labeling sperm from each boar of the pair with a different fluorophore and incubating an equal number of sperm from each boar in the same droplet with porcine oocytes. The competitive assay was highly effective in ranking boars by zona binding ability (R2=0.94). Paternity testing using microsatellite markers was used to determine the mean number of piglets sired per litter for each boar during heterospermic inseminations. The pairwise heterospermic insemination assay was effective in ranking boar fertility (R2=0.59). Using historical data from these boars, average litter size and farrowing rate were correlated (r=0.81, p<0.001). However, zona binding ability was not significantly correlated with historic farrowing rate data or historic average litter size. Boar sperm zona binding ability was also not correlated significantly with the mean number of piglets sired per litter following heterospermic insemination. But the number of piglets sired by each boar was related to a combination of zona binding ability, sperm motility, normal morphology, acrosomal integrity, and the presence of distal droplets (R2=0.70). These results suggest that zona binding ability is not an accurate predictor of fertilizing ability when used alone; however, when coupled with other sperm assessments, fertility may be predicted successfully.
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Project description:We found that factor H (FH) exists in porcine seminal plasma. Purified FH strongly inhibited serum alternative pathway complement activation against lipopolysaccharide. The molecular weight, pI, and heparin-binding activity of the purified protein were different from those of purified FH from porcine serum. The complement regulatory activity of seminal plasma FH was approximately 2-fold stronger than that of serum FH. Treatment of purified serum FH with sialidase and N-glycosidase F gave almost the same results as those of seminal plasma FH. The deletion of sialic acid from the carbohydrate chains of both FHs contributed to heparin-binding and complement regulatory activities. Results of reverse transcriptase-PCR, Western blot analysis, and immunohistochemistry showed that seminal plasma FH is mainly secreted from epithelial cells of the seminal vesicle in male genital tracts. FH was also detected in the outer acrosomal region of ejaculated sperm by immunofluorescence staining, and found that the purified FH from the sperm membrane has the same complement regulatory activity as that of seminal plasma FH. The ejaculated sperm possessing FH in the outer acrosomal region considerably evaded complement attack. We also found that there is strong complement activity in fluids from female genital tract ducts. These findings indicate that FH bound to the outer acrosomal region and soluble FH play important roles in protecting sperm against complement attack in male and female genital tracts.
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Project description:A combined Genotyping By Sequencing (GBS) and methylated DNA immunoprecipitation (MeDIP) protocol was used to identify-in parallel-genetic variation (Genomic-Wide Association Studies (GWAS) and epigenetic differences of Differentially Methylated Regions (DMR) in the genome of spermatozoa from the porcine animal model. Breeding boars with good semen quality (n = 11) and specific and well-documented differences in fertility (farrowing rate, FR) and prolificacy (litter size, LS) (n = 7) in artificial insemination programs, using combined FR and LS, were categorized as High Fertile (HF, n = 4) or Low Fertile (LF, n = 3), and boars with Unknown Fertility (UF, n = 4) were tested for eventual epigenetical similarity with those fertility-proven. We identified 165,944 Single Nucleotide Polymorphisms (SNPs) that explained 14-15% of variance among selection lines. Between HF and LF individuals (n = 7, 4 HF and 3 LF), we identified 169 SNPs with p ≤ 0.00015, which explained 58% of the variance. For the epigenetic analyses, we considered fertility and period of ejaculate collection (late-summer and mid-autumn). Approximately three times more DMRs were observed in HF than in LF boars across these periods. Interestingly, UF boars were clearly clustered with one of the other HF or LF groups. The highest differences in DMRs between HF and LF experimental groups across the pig genome were located in the chr 3, 9, 13, and 16, with most DMRs being hypermethylated in LF boars. In both HF and LF boars, DMRs were mostly hypermethylated in late-summer compared to mid-autumn. Three overlaps were detected between SNPs (p ≤ 0.0005, n = 1318) and CpG sites within DMRs. In conclusion, fertility levels in breeding males including FR and LS can be discerned using methylome analyses. The findings in this biomedical animal model ought to be applied besides sire selection for andrological diagnosis of idiopathic sub/infertility.
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Project description:Glutathione S-transferases Mu 3 (GSTM3) is an essential antioxidant enzyme whose presence in sperm has recently been related to sperm cryotolerance, quality and fertility. However, its role in seminal plasma (SP) as a predictor of the same sperm parameters has never been investigated. Herein, cell biology and proteomic approaches were performed to explore the presence, origin and role of SP-GSTM3 as a sperm quality and in vivo fertility biomarker. GSTM3 in SP was quantified using a commercial Enzyme-Linked Immunosorbent Assay (ELISA) kit specific for Sus scrofa, whereas the presence of GSTM3 in testis, epididymis and accessory sex glands was assessed through immunoblotting analysis. Sperm quality and functionality parameters were evaluated in semen samples at 0 and 72 h of liquid-storage, whereas fertility parameters were recorded over a 12-months as farrowing rate and litter size. The presence and concentration of GSTM3 in SP was established for the first time in mammalian species, predominantly synthesized in the epididymis. The present study also evidenced a relationship between SP-GSTM3 and sperm morphology and suggested it is involved in epididymal maturation rather than in ejaculated sperm physiology. Finally, the data reported herein ruled out the role of this antioxidant enzyme as a quality and in vivo fertility biomarker of pig sperm.
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Project description:In mammals, sperm migrate through the female reproductive tract to reach the egg; however, our understanding of this journey is highly limited. To shed light on this process, we focused on defining the functions of seminal vesicle secretion 2 (SVS2). SVS2(-/-) male mice produced sperm but were severely subfertile, and formation of a copulatory plug to cover the female genital opening did not occur. Surprisingly, even when artificial insemination was performed with silicon as a substitute for the plug, sperm fertility in the absence of SVS2 remained severely reduced because the sperm were already dead in the uterus. Thus, our results provide evidence that the uterus induces sperm cell death and that SVS2 protects sperm from uterine attack.
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Project description:Although there is some evidence that total dietary antioxidant capacity (TDAC) is inversely associated with the presence of obesity, no longitudinal studies have been performed investigating the effect of TDAC on comprehensive measures of body composition over time. In this study, we included 4595 middle-aged and elderly participants from the Rotterdam Study, a population-based cohort. We estimated TDAC among these individuals by calculating a ferric reducing ability of plasma (FRAP) score based on data from food-frequency questionnaires. Body composition was assessed by means of dual X-ray absorptiometry at baseline and every subsequent 3-5 years. From these data, we calculated fat mass index (FMI), fat-free mass index (FFMI), android-to-gynoid fat ratio (AGR), body fat percentage (BF%) and body mass index (BMI). We also assessed hand grip strength at two time points and prevalence of sarcopenia at one time point in a subset of participants. Data were analyzed using linear mixed models or multinomial logistic regression models with multivariable adjustment. We found that higher FRAP score was associated with higher FFMI (0.091 kg/m2 per standard deviation (SD) higher FRAP score, 95% CI 0.031; 0.150), lower AGR (-0.028, 95% CI -0.053; -0.003), higher BMI (0.115, 95% CI 0.020; 0.209) and lower BF% (-0.223, 95% CI -0.383; -0.064) across follow-up after multivariable adjustment. FRAP score was not associated with hand grip strength or sarcopenia. Additional adjustment for adherence to dietary guidelines and exclusion of individuals with comorbid disease at baseline did not change our results. In conclusion, dietary intake of antioxidants may positively affect the amount of lean mass and overall body composition among the middle-aged and elderly.
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Project description:Pulses are valued for their protein content and are good source of bioactive phytochemicals. The objectives of this study were to estimate total phenolic content (TPC), flavonoid content and total antioxidant capacity (TAC) through six different assay methods from twenty seven different varieties of pulses and split pulses commonly consumed in India. TPC and flavonoid content ranged between 38.6-542.7 mg GAE/100 g and 18.3-344.7 mg RE/100 g respectively. TAC measured by FRAP (90.6-2773.5 mg TE/100 g), RPA (190.8-2128.4 mg TE/100 g), DPPH radical scavenging assay (RSA) (42.9-571.1 mg TE/100 g), ABTS RSA (4.5-194.9 mg TE/100 g), hydroxyl RSA (0.0-106.6 mg AAE/g) and nitric oxide (NO) RSA (11.9-43.7 mg AAE/100 g) varied widely among pulses and split pulses analysed in the present study. TAC measured using different methods showed positively strong and significant correlation with TPC as well as flavonoid content except NORSA, indicating that phenolic compounds are the major contributors to the antioxidant properties of these plants. Food composition data obtained in this study would enrich the composition database at global level, which in turn will guide in improving food and nutrition security of the population. These can be used for further research, diet formulation and epidemiological studies. From the results it can be concluded that cow pea (red, small) had the highest TPC and TAC measured by all the parameters except NORSA.
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Project description:BackgroundAbnormal diet is considered to be an important risk factor for dyslipidemia. However, so far, most studies have focused on the association between single factors only, such as specific nutrients, foods, or dietary patterns, and dyslipidemia risk. This study aimed to examine the association of the joint interaction between dietary pattern and dietary total antioxidant capacity (TAC) with dyslipidemia.MethodsWe performed a dietary pattern analysis and calculated the dietary TAC based on 24-h dietary recall (DR) data from Korea National Health and Nutrition Examination Survey (KNHANES) 2007-2012, which is representative population-based cross-sectional survey in Korea. A total of 29,624 participants aged over 19?years were included for the analysis. The number of people with hypercholesterolemia, hypertriglyceridemia, and hypoHDL-cholesterolemia was 3703, 3513, and 9802, respectively. We examined the association between the joint classifications of dietary pattern score tertiles and dietary TAC level tertiles and dyslipidemia.ResultsOur results demonstrated that the "Rice & Kimchi" pattern was associated with low prevalence of hypercholesterolemia, and high prevalence of hypertriglyceridemia and hypoHDL-cholesterolemia; whereas the pattern of both "Oil, sweets, fish & other vegetables" and "Grain, bean, nuts, vegetables & fruits" were associated with low prevalence of hypertriglyceridemia. Also we demonstrated that for all dietary patterns except for the "Grain, bean, nuts, vegetables & fruits", dietary TAC was inversely associated with hypertriglyceridemia.ConclusionThis study provides basic data for the lipid-lowering effect of dietary TAC and its interaction with dietary patterns. Further study will be needed to investigate the association between dietary TAC and dietary patterns with other diseases like metabolic syndrome, cardiovascular disease, or cancer.
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Project description:Colorectal cancer is the third most common cancer worldwide. Diet has been hypothesized as involved in colorectal cancer etiology, but few studies on the influence of total dietary antioxidant intake on colorectal cancer risk have been performed.We investigated the association between colorectal cancer risk and the total antioxidant capacity (TAC) of the diet, and also of intake of selected antioxidants, in 45,194 persons enrolled in 5 centers (Florence, Naples, Ragusa, Turin and Varese) of the European Prospective Investigation into Cancer and Nutrition (EPIC) Italy study. TAC was estimated by the Trolox equivalent antioxidant capacity (TEAC) assay. Hazard ratios (HRs) for developing colorectal cancer, and colon and rectal cancers separately, adjusted for confounders, were estimated for tertiles of TAC by Cox modeling, stratifying by center.Four hundred thirty-six colorectal cancers were diagnosed over a mean follow-up of 11.28 years. No significant association between dietary TAC and colorectal cancer incidence was found. However for the highest category of TAC compared to the lowest, risk of developing colon cancer was lower (HR: 0.63; 95% CI: 0.44-0.89, P trend: 0.008). By contrast, increasing TAC intake was associated with significantly increasing risks of rectal cancer (2nd tertile HR: 2.09; 95%CI: 1.19-3.66; 3rd tertile 2.48 95%CI: 1.32-4.66; P trend 0.007). Intakes of vitamin C, vitamin E, and ß-carotene were not significantly associated with colorectal cancer risk.Further prospective studies are needed to confirm the contrasting effects of high total antioxidant intake on risk of colon and rectal cancers.
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Project description:Background: Rosehips, the fruits of Rosa species, are well known for their various health benefits like strengthening the immune system and treating digestive disorders. Antioxidant, anti-inflammatory, and cell regenerative effects are also among their health enhancing impacts. Rosehips are rich in compounds having antioxidant properties, like vitamin C, carotenoids, and phenolics. Methods: Total polyphenol content (Folin-Ciocalteu's method), and in vitro total antioxidant capacity (ferric-reducing ability of plasma, FRAP) in rosehips of four Rosa species (R. canina, R. gallica, R. rugosa, R. spinosissima) were determined and compared. Ripe fruits were harvested at two locations. Water and ethanolic extracts of dried fruit flesh were analyzed. Results:R. spinosissima had the highest total phenolic content and antioxidant capacity, significantly higher than the other investigated Rosa species. Both parameters were reported in decreasing order for R. spinosissima > R. canina > R. rugosa > R. gallica. Ethanolic extracts of rosehips showed higher phenolic content and antioxidant activity than water extracts. Antioxidant properties were influenced by the growing site of Rosa species. Conclusions: This study indicates that R. spinosissima exhibited the greatest phenolic and antioxidant content, and therefore can be used as a reliable source of natural antioxidants, and serve as a suitable species for further plant breeding activities. Furthermore, investigations of various Rosa species for their antioxidant properties may draw more attention to their potential as functional foods.
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