Antioxidant Capacity Of Seminal Plasma Research Articles

Increased Seminal Plasma Lead Levels Adversely Affect Semen Function in Painters Exposed to Lead Containing Paints.

Lead, a major contaminant around the world, is highly used in paint manufacturing due to its high anticorrosive properties. Recent reports had clearly indicated high lead content among India paints used for commercial purposes. Painters are continuously exposed to these high lead containing paints during painting of both commercial as well as house hold buildings. Lead is well known to reduce fertility in both animals as well as in humans also; we therefore aimed to measure blood lead level and its possible effect on semen functions in painters. Blood lead (BPb), activity of delta-aminolevulinc acid dehydratase (ALAD), seminal lead (SPb), parameters of semen quality, activities of antioxidants enzyme and reproductive endocrine functions in seminal plasma were measured in 103 healthy male workers, 20-40 years of age. The group contained 53 painters with slide to moderate occupational exposure to lead and 50 references subjects. All of the subjects lived in Lucknow, India. Significant (P<0.05) correlations of blood lead, delta-aminolevulinc acid dehydratase and seminal fluid lead with reproductive parameters indicated a lead related decreased in sperm density, mainly in counts of motile and viable sperm, in the percentage and count of progressively motile sperm, in antioxidant status of seminal plasma i.e. super oxide dismutase (SOD), catalase (CAT), reduced glutathione (GSH) and total antioxidant capacity of seminal plasma (TAC) where as simultaneously we also observed significant increase in abnormal sperm head and tail morphology, ROS, lipid peroxidation products (TBARS) and serum testosterone. Moreover, these associations were further confirmed by the results of multiple regression, which also showed significant (P<0.05) influence of blood lead with decreased sperm motility, antioxidant enzyme activity, TAC and increased ROS, serum testosterone. However no significant blood lead related influence was found on semen volume and liquefaction time. The seminal concentration of lead had a highly significant (P<0.01) correlation with blood lead levels (r=0.621). The overall study results indicate that even low exposure to lead (BPb<400 μg/L) can significantly reduced human semen quality with showing conclusive evidence of impairment of male reproductive endocrine functions. The use of lead in paints in India is completely unregulated and routine surveillance of paints for lead content is still lacking therefore serious consideration should be given to the urgent inclusion of regulations and bans on the use of lead in paints. (poster)

Assessment of sperm factors possibly involved in early recurrent pregnancy loss

To evaluate and compare standard sperm parameters, lipid peroxidation of sperm plasma membranes, antioxidant capacity of seminal plasma, and sperm chromatin integrity in ejaculates from men whose partners have a history of recurrent pregnancy loss and from a control group of men with recent fertility. Descriptive study. Reproduction Program, University of Antioquia, Medellín, Colombia. Twenty-three couples with history of recurrent pregnancy loss and 11 men with recent fertility. Semen samples from control men and men whose partner had a history of recurrent pregnancy loss were examined for differences in semen parameters, lipid peroxidation, antioxidant capacity, DNA fragmentation index, and sperm preparation. Migration-sedimentation method was used to collect motile spermatozoa from fresh ejaculates for examination for semen parameters. Men from the control group had spermatozoa with higher percentage of normal sperm morphology, concentration, progressive motility, and antioxidant capacity compared with men from the recurrent pregnancy loss group, who had spermatozoa with higher teratozoospermia and higher lipid peroxidation. Motile sperm fractions from both groups had spermatozoa with better sperm parameters compared with freshly ejaculated sperm. This study strengthens the current literature associating sperm quality with recurrent pregnancy loss, and emphasizes the importance of evaluating male factor by tests such as lipid peroxidation and measuring antioxidant capacity of seminal plasma in addition to conventional sperm parameters.

Effect of α-tocopherol supplementation during boar semen cryopreservation on sperm characteristics and expression of apoptosis related genes

Boar semen is extremely vulnerable to cold shock and sensitive to peroxidative damage due to high content of unsaturated fatty acids in the phospholipids of the plasma membrane and the relatively low antioxidant capacity of seminal plasma. The present study evaluated the influence of α-tocopherol supplementation at various concentrations in the boar semen extender during cryopreservation on post-thawed sperm motility characteristics (total sperm motility, MOT; local motility, LCM; curvilinear velocity, VCL; straight linear velocity, VSL; and average path velocity, VAP), sperm qualities (viability, acrosomal integrity and apoptosis), expression of stress protein (HSP70), and the expression of pro-apoptotic ( Bax and Bak) and anti-apoptotic ( Bcl-2l and Bcl-xl) genes. Semen collected from 10 Duroc boars was cryopreserved in lactose-egg yolk buffer supplemented with various concentrations of α-tocopherol (0, 100, 200, 400, 600 and 800 μM) using the straw-freezing procedure and stored at −196 °C for a minimum period of one month. In frozen-thawed groups, sperm motility was significantly ( P < 0.05) lower than that of fresh sperm. In fresh sperm, HSP70 immunoreactivity expression was observed in the equatorial region, but in frozen-thawed groups, expressions were mostly observed in the sperm head. Higher apoptosis rates were observed in 600 and 800 μM α-tocopherol supplemented frozen-thawed groups. In α-tocopherol supplemented frozen-thawed groups immediately after thawing, the expression was similar to that of fresh group. But after incubation at 37 °C for 3 h, the expression in 200 and 800 μM α-tocopherol supplemented groups was higher than that of others. Expression of pro-apoptotic genes was significantly higher and anti-apoptotic genes was significantly ( P < 0.01) lower in α-tocopherol supplemented frozen-thawed groups compared to fresh sperm group. In conclusion, α-tocopherol, supplemented at 200 μM concentration in boar semen extender during cryopreservation had a positive effect on post-thawed sperm survivability.

Influence of polymorphism of glutathione S-transferase T1 on Chinese infertile patients with varicocele

To investigate the influence of glutathione S-transferase T1 (GSTT1) gene polymorphism on Chinese infertile patients with varicocele, 63 infertile patients with varicocele and 54 healthy fertile controls were recruited in this case-control study. Our results show that oxidative damage may be the cause of infertility in patients with varicocele, and GSTT1 null genotype predisposes to over oxidative damage to spermatocytes of infertile patients with varicocele.

Attenuation of Oxidative Stress After Varicocelectomy in Subfertile Patients With Varicocele

We examined changes due to oxidative damage to spermatozoa and alterations in antioxidant capacity in subfertile patients with varicocele before and after varicocelectomy in a prospective study. A total of 30 young subfertile male patients with varicocele were recruited in this study. Varicocele was diagnosed by physical examination and Doppler ultrasound. Semen analysis was performed in the 30 patients before and 6 months after varicocelectomy using a computer assisted semen analyzer. The parameters for evaluating oxidative stress changes were 4977 bp deletion of mitochondrial DNA in sperm, as detected by polymerase chain reaction, the 8-OHdG (8-hydroxy-2'-deoxyguanosine) content in spermatozoa DNA, as measured by a high performance liquid chromatography electrochemical method, and seminal plasma protein thiols and ascorbic acid, as measured by spectrophotometric methods. Semen quality, including motility, morphology and sperm density, was improved in 22 patients (73.3%) after varicocelectomy. The incidence of 4977 bp deletion of mitochondrial DNA in sperm was 40% (12 of 30 patients) and 13.3% (4 of 30) before and after surgery, respectively. Mean +/- SD 8-OHdG content in sperm DNA, and seminal plasma protein thiols and ascorbic acid were 10.27 +/- 2.24/10(5) 2'-deoxyguanosine, 0.77 +/- 0.75 nmole/ml and 1.87 +/- 0.40 mg/dl before operation, and 5.95 +/- 1.46/10(5) 2'-deoxyguanosine, 3.00 +/- 1.17 nmole/ml and 3.12 +/- 0.94 mg/dl after surgery, respectively. The incidence of 4977 bp deletion of mitochondrial DNA in sperm and the level of 8-OHdG in sperm DNA were decreased, and seminal plasma protein thiols and ascorbic acid were increased significantly in all 30 patients after varicocelectomy. Also, in the 8 patients in whom semen quality did not improve after surgery a significant decrease in 8-OHdG in sperm DNA, and a significant increase in seminal plasma protein thiols and ascorbic acid were observed. Subfertile patients with varicocele had a significant decrease in oxidative damage in sperm DNA and an increase in antioxidant capacity in seminal plasma after varicocelectomy, indicating that surgery is effective treatment in such patients.

Antioxidant capacity of seminal plasma measured by TAS Randox®

An imbalance between reactive oxygen species (ROS) generation in sperm and antioxidant capacity of seminal plasma has been linked to male infertility. The antioxidant power of biological fluids can be evaluated either by measurement of individual antioxidants or total antioxidant capacity (TAC). The aim of this study was to assess whether TAS Randox can also be used for seminal plasma antioxidant capacity estimation. Freshly thawed aliquots of seminal plasma and blood serum of 38 males from infertile couples and 24 healthy normospermic controls were simultaneously assayed using TAS Randox reagents on the Hitachi Modular P800 Analyzer. Semen analysis was performed according to WHO guidelines. ROS in fresh sperm suspension in phosphate buffered saline was measured by chemiluminescence immediately after separation of seminal plasma. Semen analysis showed that in our study group only 14 males were normospermic and 24 males had mostly combined pathologies. The medians for ROS production were similar in both the study and the control groups (4850 and 5450 RLU/min, resp). Seminal plasma TAS levels were significantly lower (p<0.02) in the study group while blood serum TAS levels were similar in both groups. A significant positive correlation (p<0.05) between TAS in seminal plasma and serum was found, seminal plasma levels being on average 1.4 times higher. The TAS Randox kit may be used for clinical studies intended to identify decreased antioxidant power in the seminal fluid of infertile men.

The effects of dietary lycopene supplementation on human seminal plasma

To investigate whether lycopene levels in blood and seminal plasma increase after dietary supplementation with a natural source of the compound, and whether any potential increase of lycopene levels in semen translates into increased free-radical trapping capacity in the seminal plasma. Reactive oxygen species are detrimental to the health and function of spermatozoa. Semen contains enzymatic and non-enzymatic defence mechanisms to combat such species, and lycopene, a dietary antioxidant, forms part of the non-enzymatic arm. Immuno-infertile men have significantly lower levels of lycopene in their semen, and oral lycopene therapy can improve various seminal variables in idiopathic infertility. Whether this improvement is a direct consequence of increased lycopene levels in semen, resulting in an increased radical scavenging ability, remains unknown. Blood and seminal lycopene levels were measured in healthy volunteers, using high-performance liquid chromatography, before and after a period of dietary supplementation. The antioxidant capacity of seminal plasma was also assayed to determine if supplementation results in a measurable increase in seminal radical scavenging ability. There were statistically significant increases in blood and seminal plasma lycopene levels after dietary supplementation. The increase in seminal and blood lycopene levels showed a strong positive correlation (r = 0.84, P < 0.05). There was no measurable increase in the total radical scavenging capacity of semen. This study confirms the presence of lycopene in human semen, the levels of which can be significantly increased after dietary supplementation with a natural source of lycopene. Further studies to establish whether this would also be the case in infertile men, with possible associated improvements in their seminal quality, are warranted.

Increased Total Antioxidant Capacity in Seminal Plasma of Varicocele Patients: A Multivariate Analysis

To investigate seminal antioxidant systems and their correlation with hormonal pattern in varicocele patients, we studied 33 varicocele (VAR) patients (12 oligozoospermic, 21 normozoospermic) and 34 non-VARs (10 idiopathic oligozoospermic, 24 normozoospermic). Non-enzymatic total antioxidant capacity (TAC) was measured using H(2)O(2)-metamyoglobin, which generates the radical form, spectroscopically detectable, of the chromogen 2,2',-azinobis-3-ethylbenzothiazoline-6-sulphonate (ABTS): time till appearance (Lag) of ABTS*(-) signifies antioxidant concentration. Lag was significantly longer in VARs than controls suggesting ineffective utilization of antioxidants. A significant direct correlation (r = 0.65, p < 0.01) of Lag with sperm count was observed in non-VARs, while in VARs it was inverted, as well as with hematic FSH levels. A multivariate analysis including FSH, Lag, progressive spermatozoa, oligozoospermia and varicocele indicated a strong inverse correlation between FSH and motility (r(2) = 0.31, p > F = 0.0007), not modified by Lag (r(2) = 0.31, p > F = 0.002). Their inverse correlation with Lag may suggest that higher FSH levels, improving sperm antioxidant efficiency, counterbalance varicocele-induced dyspermia.

1063 Automated measurement of total antioxidant capacity of seminal plasma

1063 Automated measurement of total antioxidant capacity of seminal plasma

Coenzyme Q10 effects on non-enzymatic total antioxidant capacity in seminal plasma of varicocele patients

In order to investigate antioxidant systems in varicocele (VAR), we studied the correlation between non-enzymatic total antioxidant capacity (TAC) and seminal parameters and subsequently the effect of supplementation of antioxidant, coenzyme Q10 (CoQ10). TAC was determined in seminal plasma of 27 VAR patients (10 oligospermic and 17 normospermic) and 24 non-VAR controls (7 with idiopathic oligospermia and 17 normospermic). We measured the interaction of system H 2O 2-metamyoglobin with chromogen ABTS, whose radical cation is spectroscopically detectable. The length of latency phase (Lag) in ABTS accumulation is proportional to antioxidant concentration. Lag was significantly greater in all VAR vs. control subjects; a multivariate analysis indicated a very high inverse correlation between FSH and the percentage of motile progressive cells, which was not further modified by addition of Lag length to the statistical model. CoQ10 treatment (100 mg/daily for 3 months) significantly increased the Lag phase. We maintain that greater FSH levels, achieved by inducing a more efficient utilisation of antioxidants by spermatozoa, may counterbalance VAR-induced spermatozoa deficit.

Relationship between seminal ascorbic acid and sperm DNA fragmentation in infertile men

Oxidative stress, a well known cause of abnormal semen parameters, may also impact sperm DNA fragmentation. Ascorbic acid has been reported to have a protective effect on sperm DNA fragmentation induced by exogenous oxidative stress in vitro. But, there is no report of beneficial effects of ascorbic acid on sperm DNA fragmentation in infertile men. The aim of this study is to investigate the correlation between measures of seminal plasma antioxidant capacity and sperm DNA fragmentation in infertile men.

Varicocelectomy reduces reactive oxygen species levels and increases antioxidant activity of seminal plasma from infertile men with varicocele.

Several theories have been advanced to explain the mechanisms by which varicocele impairs male fertility. These theories include scrotal hyperthermia, retrograde flow of adrenal or renal metabolites, Leydig cell dysfunction and hypoxia. Varicocele is reported to be associated with elevated reactive oxygen species (ROS) production in spermatozoa and diminished seminal plasma antioxidant activity. The aim of this study was to investigate whether surgical correction of varicocele might reduce ROS or increase the antioxidant capacity of seminal plasma from infertile patients with varicocele. The study group consisted of 68 infertile males, selected from patients scheduled for varicocelectomy at Cairo University Hospital during the year 1999. Seminal plasma levels of two ROS [malondialdehyde (MDA), hydrogen peroxide (H2O2)] and one ROS radical [nitric oxide (NO)] were estimated as well as six antioxidants [superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), vitamin C (Vit C), vitamin E (Vit E), albumin) on the day prior to varicocelectomy. For comparison, the same parameters were measured again 3 and 6 months post-operatively. A statistically significant reduction in the 3 month post-operative levels of MDA, H2O2 and NO was observed when compared with the pre-operative values. A further significant reduction took place during the following 3 months. Four of the six antioxidants tested (SOD, CAT, GPx, and Vit C) showed a significant increase in seminal levels when comparing 3-month post-operative with pre-operative values. A further significant increase of the four antioxidant levels took place during the following 3 months. No significant change in the level of seminal plasma albumen took place during the first 3 months after varicocelectomy, however, a significant increase was noted during the next 3 months. In contrast to other antioxidants, seminal plasma levels of Vit E showed a significant decrease when comparing 3-month post-operative with pre-operative values. A further significant decrease took place during the following 3 months. It is concluded that varicocelectomy reduces ROS levels and increases antioxidant activity of seminal plasma from infertile men with varicocele.

Impaired seminal antioxidant capacity in human semen with hyperviscosity or oligoasthenozoospermia.

Antioxidant capacity of seminal plasma was evaluated in 120 semen samples subdivided into asthenozoospermic and oligoasthenozoospermic specimens with normal consistency and into asthenozoospermic and oligoasthenozoospermic specimens with hyperviscosity. Semen samples (n = 25) from normozoospermic donors were used as a control group. Scavenger antioxidant capacity of reactive oxygen species was evaluated by superoxide dismutase and catalase activity measurements, whereas the chain-breaking antioxidant efficiency was detected by total antioxidant status assessment. In semen with normal viscosity, unaltered enzymatic and nonenzymatic antioxidant capacity was revealed in the asthenozoospermic specimens, whereas low superoxide dismutase activity was detected in oligoasthenozoospermic samples. On the contrary, impairment of both the scavenger and chain-breaking antioxidative systems was revealed in asthenozoospermic and oligoasthenozoospermic hyperviscous ejaculates, regardless of sperm count. Catalase activity and total antioxidant status values were also reduced in the 2 subgroups of hyperviscous ejaculates compared with their respective matched controls, whereas similar superoxide dismutase activities were detected in oligoasthenozoospermic samples with normal and high consistencies. These results suggest that asthenozoospermia could be related to an antioxidant deficiency only in combined ejaculate pathologies, and that a severe impairment of the low and high molecular weight seminal antioxidative capacities could be associated with semen hyperviscosity.

Oxidative damage to proteins and decrease of antioxidant capacity in patients with varicocele

To examine oxidative damage to blood proteins in the spermatic vein and seminal plasma antioxidant capacity of patients with varicocele, 30 young male patients with varicocele (group 1), 25 young male patients with subclinical varicocele (group 2), and 15 normal young males without varicocele (group 3) were recruited in this study. Varicocele and subclinical varicocele were confirmed by physical examination and Doppler ultrasonography. Blood samples were drawn from peripheral and spermatic veins before varicocelectomy. Plasma protein carbonyls were measured by a spectrophotometric assay after reacting with 2,4-dinitrophenylhydrazine. Protein thiols and ascorbic acid of seminal plasma were measured by spectrophotometric methods. We found that plasma protein carbonyls in the spermatic veins were significantly higher than those of corresponding peripheral veins in all 30 patients in group 1 and 12 patients in group 2 receiving varicocelectomy. Protein carbonyls in the spermatic veins of patients with varicocele (3.72 ± 0.56 nmole/mg protein) and patients with subclinical varicocele (3.50 ± 0.30 nmole/mg protein) were found to be higher than those of the control (2.35 ± 0.33 nmole/mg protein). Protein thiols were 0.97 ± 0.96, 1.50 ± 0.89, and 3.49 ± 0.81 nmole/ml, and ascorbic acid levels were 1.87 ± 0.42, 2.13 ± 0.24, and 2.38 ± 0.07 mg/dl, in seminal plasma of the patients in groups 1, 2, and 3, respectively. Seminal plasma protein thiols and ascorbic acid levels in group 1 were significantly lower than those in groups 2 and 3, respectively. These results indicate that oxidative stress in the patients with varicocele and subclinical varicocele was higher than that of the control. We suggest that plasma protein carbonyls, and protein thiols and ascorbic acid of seminal plasma are useful markers for the assessment of oxidative stress in patients with varicocele and subclinical varicocele.

VARICOCELECTOMY IMPROVES INTRAUTERINE INSEMINATION SUCCESS RATES IN MEN WITH VARICOCELE

We determined whether varicocele treatment before intrauterine insemination significantly affects intrauterine insemination success rates. A total of 58 infertile couples, of whom the women had normal evaluations and men had abnormal semen analyses and a history of varicocele, were included in this study. They were identified after reviewing the charts of all women undergoing intrauterine insemination for male factor infertility at our center. Of the men 24 participated in 63 intrauterine insemination cycles without varicocele treatment, while in the remaining 34 varicocele was treated before a total of 101 intrauterine insemination cycles. Variables associated with pregnancy or live birth were analyzed using repeat measures logistic regression with generalized estimating equation techniques. An initial stepwise generalized estimating equation was performed without including varicocele treatment status. Subsequently varicocele treatment status and the significant associated factors were included in analysis. The semen characteristics of untreated and treated varicocele groups were compared with repeat measures analysis of variance. On pre-wash semen analysis patients with untreated varicocele had significantly higher mean motility plus or minus standard error than patients whose varicoceles were treated (48.6% +/- 2.3% versus 38.1% +/- 1.8%, p = 0.02). However, no statistically significant difference was noted in the mean post-wash total motile sperm count in the treated and untreated groups (7.2 +/- 1.0 versus 14.8 +/- 2.6, p = 0.1). Despite these findings the pregnancy and live birth rates per cycle were significantly higher in patients in whom varicocele was treated than in those without varicocele treatment (11.8% versus 6.3%, p = 0.04 and 11.8% versus 1.6%, p = 0.007, respectively). Varicocele treatment may not improve semen characteristics in all men but it appears to improve pregnancy and live birth rates among couples undergoing intrauterine insemination for male factor infertility. A functional factor not measured on routine semen analysis may affect pregnancy rates in this setting. Men should be screened for varicocele before intrauterine insemination is initiated for male factor infertility.

Tyrosine as important contributor to the antioxidant capacity of seminal plasma

A novel post-addition method, based on the trapping of ABTS-radicals, is applied for studying the total antioxidant capacity of seminal plasma. A remarkable profile is observed, in which seminal plasma quenches radicals in a continuous, relatively slow fashion. Five putative antioxidants present in seminal plasma were studied using the same assay. Some of the compounds such as ascorbic acid, α-tocopherol and uric acid exert immediate, fast radical trapping, whereas hypotaurine and tyrosine give rise to the same slow radical trapping curve as seminal plasma. Due to this slow, continuous radical trapping, quantification of the total antioxidant capacity (expressed as trolox equivalent antioxidant capacity, TEAC) strongly depends on the chosen time point after onset of radical trapping. When determined during the slow antioxidant trapping phase, tyrosine has a powerful antioxidant capacity, which in combination with its relatively high plasma concentration makes it an important contributor to the total antioxidant capacity of seminal plasma.

ASSOCIATION OF UREAPLASMA UREALYTICUM WITH ABNORMAL REACTIVE OXYGEN SPECIES LEVELS AND ABSENCE OF LEUKOCYTOSPERMIA

Ureaplasma urealyticum is a commensal of the lower genitourinary tract of many sexually active adults. The organism is more common in partners of infertile than fertile marriages. We conducted a prospective study at our tertiary care center to confirm a possible association between U. urealyticum and abnormal sperm function parameters. A total of 50 consecutive male patients seeking general urology consultation for lower urinary tract symptoms characteristic of chronic prostatitis were evaluated. Urine and semen localization cultures were performed with additional semen cultures for U. urealyticum, Chlamydia trachomatis and Mycoplasma hominis. Specimens from 21 healthy men were used as controls. Specimens were analyzed by a computer assisted semen analyzer, and verified manually for concentration, percent motility and morphology. Leukocytospermia was measured by the Endtz test. Semen specimens were also analyzed for reactive oxygen species (ROS), acrosome reaction and mannose binding assay. Of the patients 17 had positive U. urealyticum cultures and the other cultures were negative. Patients with U. urealyticum had significantly higher ROS levels (log [ROS + 1] = 2.52 +/- 0.25) than those without U. urealyticum (1.49 +/- 0.20, p = 0.002) or controls (1.31 +/- 0.19, p = 0.002). Leukocytospermia was detected in only 1 of the 17 (6%) positive specimens and 4 (12%) negative specimens. Seminal ROS levels are elevated among patients with U. urealyticum. ROS induces lipid peroxidation, which reduces membrane fluidity and sperm fertilization capability, and may be the mechanism by which U. urealyticum impairs sperm function. Absence of leukocytospermia does not exclude U. urealyticum.

Total antioxidant capacity of seminal plasma is different in fertile and infertile men

To determine if the total antioxidant capacity of seminal plasma is different in fertile and infertile men. An enhanced chemiluminescence assay applied to seminal plasma from groups of fertile and infertile men. The Assisted Conception Unit, Royal Maternity Hospital, Belfast. Men of proven fertility whose partners had an ongoing pregnancy resulting from IVF and male partners of couples attending our subfertility clinic. Total antioxidant capacity was significantly higher in seminal plasma from fertile men than from that of infertile men with normozoospermic samples that exhibited reactive oxygen species or asthenozoospermic samples with or without reactive oxygen species activity. Seminal plasma from infertile men has lower antioxidant levels than that of fertile men, particularly of patients whose semen have poor sperm motility. The presence of reactive oxygen species activity in sperm of infertile groups also is associated with lower levels of chain-breaking antioxidants in seminal plasma.