Significance of malondialdehyde concentration in seminal plasma of infertile men

Objective: To determine the concentration of malondialdehyde (MDA), product of lipid peroxidation, in seminal plasma and evalu ate its significance. Methods: Ninety-three cases of infertile pa tients were divided into the obstructive azoospermic (12 cases), the non-obstructive azoospermic (15 cases), the oligozoospermic (21 cases), the asthenozoospermic (19 cases), the oligoastheno zoospermic (16 cases) and the oligoasthenoteratozoospermic (10) groups. Eighteen fertile males served as the controls. MDA con centration was assessed by high-performance liquid chromatogra phy (HPLC). Results: With the exception of the obstructive azoospermic group, the MDA concentration in the seminal plasma was significantly different between the control group and the infer tile groups (P<0.01) as well as between the different infertile groups. Conclusion: Determination of MDA concentration in the seminal plasma is helpful to the diagnosis of male infertility induced by overproduction of reactive oxygen species.     

Studies on relationship between semen viscosity and other semen parameters, Ureaplasma urealyticum infection and seminal plasma antisperm antibody

Objective: To study the relationship between semen viscosity and other semen parameters, Ureaplasma urealyticum (UU) infection and seminal plasma antisperm antibody (AsAb) in male infertiles. Methods: Semen parameters, Ureaplasma urealyticum (UU) infection and antisperm antibody (AsAb) were measured and analyzed in 4337 infertile men. Results: The seminal viscosity was higherr than normal in 65.02 % of 4337 male infertiles. The sperm motility and grade (a, b) motile sperm were significantly lower in the high viscosity group than in the normal viscosity group (P<0.05-0.01). The rate of abnormal morphology sperm was higher and duration of semen liquefaction was longer in the high viscosity than in the normal viscosity group (P<0.01). The seminal volume, sperm concentration and semen pH were not significantly different between the two groups. The semen viscosity is significantly higher in subjects with higher seminal WBC (>5/ HP) than in those with lower WBC (<5/HP). The positive AsAb and UU infection rates     

Do malignant diseases affect semen quality? Sperm parameters of men with cancers

The advent of modern treatments together with the improvement of the surgical techniques has significantly increased 5‐year survival rates of young patients with cancer. Although the deleterious effects of chemotherapy and radiation are well documented, controversies exist about the effect of cancer itself on semen parameters before treatment. We collected data on 236 patients representative of different types of cancers reoffered at our institution for sperm cryopreservation with the aim to correlate the pre‐freeze semen parameters with type of cancer, disease stage and with semen quality of 102 fertile and healthy men. The median baseline semen parameters of all our patients with cancer are placed above the 5th percentile of the World Health Organization reference value, but the type of cancer may impact the sperm parameters. In testicular tumours and in Hodgkin lymphoma, we show a semen concentration statistically lower than in the fertile population, while in patients with other cancers, there is no difference with the healthy men. We found no correlation between semen quality and disease stage. Eighty‐six per cent of our patients do not have children at the time of semen cryopreservation, and the only established clinical option for preserving fertility of these men is cryopreservation of spermatozoa.     

Studies on sperm forward motility-elated proteins in seminal plasma: isolation and activity detecion

Employing techniques of the affinity chromatography coveredwith ConA beads and protein ultrafiltration, certain proteins wereisolated and concentrated from the heated human seminal plasma.The results of computer assistant semen analyses (CASA) revealedthat the forward motility index of spermatozoa from bovine epididy-mal caput incubated with proteins and theophylline was significantlyhigher than that incubated with theophylline only (P < 0.01). Us-ing the methods of PAGE, SDS-PAGE and Coomassie blue stain-ing, the protein bands with high molecules in the gel of PAGEchanged into low molecules in the gel of SDS-PAGE. The majorbands of the proteins in the gel of SDS-PAGE were 46.8 and 57.5kDa. The proteins related to the forward motility of spermatozoa inmammalian seminal plasma play vital roles during sperm maturationin the epididymis and fertilization. (Chin J Androl 2001; 2: 87-91)     

Does sperm morphology affect the outcome of intrauterine insemination in patients with normal sperm concentration and motility?

The aim of this study was to assess the correlation of sperm morphology with the intrauterine insemination (IUI) outcome in patients with normal sperm concentration and motility. About 412 couples who underwent 908 IUI cycles were involved in the present study. A total of 110 clinical pregnancies were achieved with a pregnancy rate of 12.11% per cycle. The pregnancy rates per cycle were 7.60%, 12.67%, 13.62% and 13.13% in patients with <5%, 5–9%, 10–14% and >14% normal forms, respectively. The lowest pregnancy rate (7.60%) was obtained in the group with normal forms below 5%. However, this rate was not significantly different from other subgroups. Moreover, no pregnancies occurred in women >35 years old with normal sperm forms below 5%, in comparison with that in other subgroups of the same age. For women younger than 35 years old, no significant difference in pregnancy rate was observed in terms of different level of morphologically normal sperm. Our results show that for patients with normal sperm concentration and motility, IUI is recommended for first‐line treatment when the woman is younger than 35 years, or morphologically normal sperm is ≥5%. IVF/ICSI should be performed when the normal forms are <5% and female age is >35 years.     

Seminal plasma anti-Müllerian hormone level correlates with semen parameters but does not predict success of testicular sperm extraction (TESE)

AIM: To assess seminal plasma anti-Müllerian hormone (AMH) level relationships in fertile and infertile males. METHODS: Eighty-four male cases were studied and divided into four groups: fertile normozoospermia (n = 16), oligoasthenoteratozoospermia (n = 15), obstructive azoospermia (OA) (n = 13) and non-obstructive azoospermia (NOA) (n = 40). Conventional semen analysis was done for all cases. Testicular biopsy was done with histopathology and fresh tissue examination for testicular sperm extraction (TESE) in NOA cases. NOA group was subdivided according to TESE results into unsuccessful TESE (n = 19) and successful TESE (n = 21). Seminal plasma AMH was estimated by enzyme linked immunosorbent assay (ELISA) and serum follicular stimulating hormone (FSH) was estimated in NOA cases only by radioimmunoassay (RIA). RESULTS: Mean seminal AMH was significantly higher in fertile group than in oligoasthenoteratozoospermia with significance (41.5 +/- 10.9 pmol/L vs. 30.5 +/- 10.3 pmol/L, P < 0.05). Seminal AMH was not detected in any OA patients. Seminal AMH was correlated positively with testicular volume (r = 0.329, P = 0.005), sperm count (r = 0.483, P = 0.007), sperm motility percent (r = 0.419, P = 0.021) and negatively with sperm abnormal forms percent (r = -0.413, P = 0.023). Nonsignificant correlation was evident with age (r = -0.155, P = 0.414) and plasma FSH (r = -0.014, P = 0.943). In NOA cases, seminal AMH was detectable in 23/40 cases, 14 of them were successful TESE (57.5%) and was undetectable in 17/40 cases, 10 of them were unsuccessful TESE (58.2%). CONCLUSION: Seminal plasma AMH is an absolute testicular marker being absent in all OA cases. However, seminal AMH has a poor predictability for successful testicular sperm retrieval in NOA cases.     

Does folic acid and zinc sulphate intervention affect endocrine parameters and sperm characteristics in men?

We evaluated pre- and post-intervention endocrine and semen parameters in a double-blind, placebo-controlled intervention study to investigate the underlying mechanism of increased sperm concentration after folic acid and zinc sulphate intervention. A total of 47 fertile and 40 subfertile males participated in a 26-week intervention study consisting of a daily treatment with folic acid (5 mg/day) and zinc sulphate (66 mg/day), or placebo. Pre- and post-intervention semen parameters, serum folate, zinc, follicle-stimulating hormone (FSH), testosterone and inhibin B concentrations were measured. The results indicated that intervention treatment significantly increased sperm concentration in subfertile males. Other semen and endocrine parameters were not affected by intervention treatment. At baseline, positive correlations were found between serum zinc and sperm concentration, motility and inhibin B. Serum zinc and FSH were inversely correlated. As (already) well known from previous research, inhibin B positively correlated with sperm concentration, motility and morphology, and was inversely correlated with FSH. The latter was positively correlated with testosterone. In addition, testosterone and inhibin B were inversely correlated. After intervention, the correlations with zinc disappeared. We conclude that the increase in sperm concentration after folic acid and zinc sulphate intervention is not the result of alterations in FSH, testosterone or inhibin B concentrations. Although zinc and folate have several effects on spermatogenesis, the underlying mechanisms involved are not clear.     

Analyis of Protamine mRNA in sperm of men,rats and mice

Aim: To investigate the existence of the protamine mRNA insperm of men, rats and mice. Methods: By means of RT-PCRtechniques, protamine cDNA fragments were obtained from totalRNA of the mature sperm of men, rats and mice. Results:mRNA of protamine gene was present in the mature sperm of men,rats and mice. The protamine cDNA obtained by RT-PCR in ratsperm with an abnormal head was much less in number than that inthe normal rat sperm. Conclusion: mRNA in the sperm mightrepresent the condition of corresponding gene expression duringspermatogenesis. (Reprod Contracep 2001; 21: 200 - 5)     

Does varicocelectomy improve semen in men with azoospermia and clinically palpable varicocele?

The effectiveness of varicocelectomy in nonobstructive azoospermia is controversial. The current study assessed the efficacy of microsurgical subinguinal varicocelectomy in nonobstructive azoospermic men with palpable varicocele and to evaluate predictive parameters of outcome. We reviewed the records of 723 patients who had microsurgical varicocelectomy and diagnostic testicular biopsy between 2012 and 2016 at a tertiary medical centre. Data pertaining to the physical, laboratory (semen analysis and hormonal profile) and histopathology features were examined, exploring the predictors of improvement in semen analysis post-varicocelectomy. In total, 42 patients with mean age 35.71 ± 6.35 years were included. After a mean varicocelectomy follow-up of 6.7 months, motile spermatozoa in the ejaculate could be observed in 11 patients (26.2). Out of all the factors examined, only testicular histopathology significantly predicted post-varicocelectomy outcome, where 8/11 patients exhibited hypospermatogenesis, and 3/11 Sertoli cell-only regained spermatozoa in semen. Microsurgical varicocelectomy in nonobstructive azoospermic men with clinically palpable varicocele can result in sperm appearance in the ejaculate with the highest success expected in hypospermatogenesis.     

Seminal plasma anti-Müllerian hormone level correlates with semen parameters but does not predict success of testicular sperm extraction （TESE）

Aim： To assess seminal plasma anti-Müllerian hormone （AMH） level relationships in fertile and infertile males. Methods： Eighty-four male cases were studied and divided into four groups： fertile normozoospermia （n = 16）, oligoastheno- teratozoospermia （n = 15）, obstructive azoospermia （OA） （n = 13） and non-obstructive azoospermia （NOA） （n = 40）. Conventional semen analysis was done for all cases. Testicular biopsy was done with histopathology and fresh tissue examination for testicular sperm extraction （TESE） in NOA cases. NOA group was subdivided according to TESE results into unsuccessful TESE （n = 19） and successful TESE （n = 21）. Seminal plasma AMH was estimated by enzyme linked immunosorbent assay （ELISA） and serum follicular stimulating hormone （FSH） was estimated in NOA cases only by radioimmunoassay （RIA）. Results： Mean seminal AMH was significantly higher in fertile group than in oligoasthenoteratozoospermia with significance （41.5±10.9 pmol/L vs. 30.5±10.3 pmol/L, P 〈 0.05）. Seminal AMH was not detected in any OA patients. Seminal AMH wascorrelated positively with testicular volume （r = 0.329, P = 0.005）, sperm count （r = 0.483, P = 0.007）, sperm motility percent （r = 0.419, P = 0.021） and negatively with sperm abnormal forms percent （r = -0.413, p = 0.023）. Nonsignificant correlation was evident with age （r = -0.155, P = 0.414） and plasma FSH （ r = -0.014, P = 0.943）. In NOA cases, seminal AMH was detectable in 23/40 cases, 14 of them were successful TESE （57.5%） and was undetectable in 17/40 cases, 10 of them were unsuccessful TESE （58.2%）. Conclusion： Seminal plasma AMH is an absolute testicular marker being absent in all OA cases. However, seminal AMH has a poor predictability for successful testicular sperm retrieval in NOA cases.     

Semen parameters in men with spinal cord injury： changes and aetiology

Aim： To assess the changes in semen parameters in men with spinal cord injury （SCI） and the possible causes of these changes. Methods： The study included 45 subjects with SCI. Semen retrieval was done by masturbation （2）, vigorous prostatic massage （n = 13）, penile vibratory stimulation （n = 13） or electroejaculation （n = 17）. Results： The semen of men with SCI showed normal volume （2.3 ± 1.9 mL） and sperm count （85.0 × 10^6 ± 83.8 × 10^6/mE） with decreased motility （11.6% ± 10.1%）, vitality （18.5 % ± 15.2%） and normal forms （17.5 ± 13.4%）, and pus cells has been increased （6.0 × 10^6 ± 8.2 × 10^6/mL）. Total （13.4 ± 9.9 vs. 7.1 ± 6.8） and progressive （4.4 ± 3.9 vs. 2.2 ± 2.1） motility were significantly higher in subjects with lower scrotal temperatures. There was no statistical significant difference between electroejaculation and penile vibratory stimulation groups as regards any of the semen parameters. Subjects＇ age, infrequent ejaculation, injury duration and hormonal profile showed no significant effect on semen parameters. Conclusion： The defining characteristics of the seminogram in men with SCI are normal volume and count with decreased sperm motility, vitality and normal forms, and the increased number of pus cells. The most acceptable cause of the deterioration of semen is elevated scrotal temperature.     

Distribution of mouse interferon-β in normal and brain tumour-bearing mice

Summary The distribution of¹²⁵I-labelled recombinant mouse interferon- (rMuIFN-) in normal and glioma (203 glioma) bearing mice was studied by radioassay and macro-autoradiography at 15 and 30 min after a single intravenous injection. The level of rMuIFN- in the spleen was about 20-fold higher than in serum. Concentrations higher than the serum level was detected in the lung, liver and kidney. The concentration of rMuIFN- in the brain was 8% of the serum level and the concentration in the glioma 30 min after administration was about 10-fold higher than in normal mouse brain. Macro-autoradiographic study demonstrated a wide distribution range and selective uptake in glioma tissue. Furthermore, we found that mouse gliomas were sensitive to mouse IFN-. Our findings demonstrate that in the mouse glioma model, intravenously administered interferon reaches the tumour.     

May antioxidant therapy improve sperm parameters of men with persistent oligospermia after retrograde embolization for varicocele?

We performed a randomized, prospective, controlled, intention to treat study in order to determine the effectiveness of an antioxidant therapy in improve the quality of seminal fluid parameters and the natural pregnancies in men with persistent oligospermia (5–20 million/ml) 6 months after retrograde embolization. Forty-two subjects were enrolled and randomized in the study. Treated group (20 subjects) was assigned to receive antioxidant therapy (NAC 600 mg and vitamins–minerals). Untreated group (22 subjects) received no adjunctive medical therapy and was used as controls. Our data were analyzed with an intention to treat strategy. A statistically significant increase in sperm count after antioxidant therapy was recorded (P = 0.009). After this therapy, no statistical differences in percentage of WHO class A motile sperm (P = 0.752) and typical forms (P = 0.926) were found. The univariate logistic regression analysis showed that a man treated with antioxidant therapy presented a probability to have a normal sperm count 20-fold (OR = 20.1; CI 95% = 1.05–43.2; P = 0.014) higher than a man who was untreated. No significant impact on spontaneous pregnancies was found after antioxidant therapy. Despite this preliminary data, we show that antioxidant therapy based on a combination of NAC and micronutrient supplementation can be helpful in improve the sperm count at least in a subset of oligospermic males. However, this improving in sperm count is not associated with a significant increase in spontaneous pregnancies after 12 months.     

Combined effect of DHA and α-tocopherol supplementation during bull semen cryopreservation on sperm characteristics and fatty acid composition

The aim of this study was to investigate the effect of adding n-3 fatty acids (FA) and α-tocopherol (VE) to semen extender on freezing ability and FA composition of bull sperm. Semen was collected from 10 Iranian Holstein bulls and was pooled. In the first experiment, semen was divided into 12 groups including 4 levels of n-3 FA (0, 0.1, 1, 10 ng ml(-1) ) and 3 levels of VE (0, 0.1, 0.2 mmol). The treatment of 0.2 mmol VE and 10 ng ml(-1) n-3 FA had the best post-thawed sperm characteristics (P < 0.01). In the second experiment, lipid composition of the latest treatment and control (without FA and VE) was determined. Adding n-3 FA increased docosahexaenoic acid (DHA) percentage before freezing and after thawing. The ratio of n-3 to n-6 before freezing was higher (P < 0.05) in treated group than in control, and this ratio in the fresh sperm was greater than in the post-thawed sperm (P = 0.1). Results suggested that adding DHA accompanied with an antioxidant to an extender could improve cryosurvival of bull sperm via altering membrane lipid composition.     

Association of XRCC1 and ERCC2 promoters’ methylation with chromatin condensation and sperm DNA fragmentation in idiopathic oligoasthenoteratozoospermic men

The aim of the study was to investigate whether the promoter methylation of XRCC1 and ERCC2 genes is associated with sperm DNA fragmentation and chromatin condensation in idiopathic oligoasthenoteratozoospermic men. This study involved 77 infertile men with idiopathic oligoasthenoteratozoospermia and 51 normozoospermic controls. The methylight method, TUNEL assay and aniline blue staining were used for the evaluation of XRCC1 and ERCC2 genes’ methylation, SDF and sperm chromatin condensation, respectively. SDF (p = .004) and XRCC1 methylation (p = .0056) were found to be significantly higher in men with idiopathic OAT than in the controls, while mature spermatozoa frequency was higher in controls as compared to infertile men (p < .0001). No significant association was found between SDF and methylation of XRCC1 and ERCC2 genes (p = .9277 and p = .8257, respectively). However, compared to the cut-off point obtained by receiver operating characteristic analysis, a significant association was found between SDF and XRCC1 methylation, positive and negative methylation groups, generated according to the cut-off value for XRCC1. XRCC1 methylation was found to have a significant effect on chromatin condensation (p = .0017). No significant difference was detected among ERCC2 methylation, male infertility and SDF. In conclusion, XRCC1 methylation may have a role in sperm chromatin condensation and idiopathic OAT.     

Quick recovery and characterization of cell-free DNA in seminal plasma of normozoospermia and azoospermia： implications for non-invasive genetic utilities

We established a quick and reliable method for recovering cell-free seminal DNA （cfsDNA）, by using the binding-washing-elution procedure on the DNA purification column. Low variations （below 15%） among the triplicate values of cfsDNA quantity verified the reproducibility of our cfsDNA recovery method. Similar cfsDNA yield and size distribution between seminal plasma acquired by filtration and centrifugation confirmed the presence of cfsDNA. To investigate the general characterization of cfsDNA, the quantitation and size distribution of cfsDNA from normozoospermic and azoospermic semen were analyzed by real-time PCR and electrophoresis, respectively. CfsDNA concentration in semen with normozoospermia （n = 11） was 1.34 ± 0.65 μg ·mL^-1, whereas a higher cfsDNA concentration was observed in azoospermia （2.56 ± 1.43 μg ·mL^-1, n = 9）. The continuous distribution of DNA fragments ranging from -1 kb to 15 kb and a spectrum of multiples of 180-bp fragments were observed in each normozoospermic and azoospermic sample. Distinct characteristic DNA ladder fragmentations in some azoospermic samples implicated that cfsDNA originate partly from apoptotic cells. CfsDNAs of 36 selected azoospermic patients with known information of Y chromosome microdeletion were subjected to the same microdeletion analysis by multiplex PCR and PCR amplification of sY114 （1 450 bp）. All multiplex PCR reactions with cfsDNA amplified successfully and provided the same result as leukocyte DNA. PCR amplification of sY114 gave a 1 450-bp amplicon as expected. Our data suggested the potential use of cfsDNA in search of biomarker or diagnostic procedures.     

Is there any correlation between sperm parameters and chromatin quality with embryo morphokinetics in patients with male infertility?

The main goal was to evaluate the correlation between sperm parameters and chromatin quality with embryo kinetics via time‐lapse monitoring system (TLM). A total of 40 couples involved in the ICSI program as a result of male infertility. For assessment of sperm chromatin and DNA quality, we used aniline blue, toluidine blue, chromomycin A3, acridine orange and terminal transferase‐mediated deoxyuridine triphosphate biotin end labelling assays. All mature oocytes were injected, and the generated zygotes (2PNs) were cultured in TLM. In day 3 after injection, single embryo transfer (SET) was carried out according to the morphology and morphokinetics. The patients were followed up until delivery. There were positive significant correlations between sperm count with CC2 (r = .330, p = .049), T4 (r = .329, p = .038), T6 (r = .342, p = .035) and T7 (r = .374, p = .025). Also, there were positive significant correlations between nonprogressive motility and T2 (r = 0.323, p = .042), T3 (r = .411, p = .013) and T4 (r = .418, p = .007). Regarding the sperm chromatin quality assays, there were negative significant correlations between CMA3 and CC2 (r = ?.272, p = .049) and between acridine orange and T5 (r = ?.221, p = .040). It seems that the abnormal sperm parameters and chromatin alteration affect the normal embryo kinetics in ICSI program.     

High levels of oxidation–reduction potential in frozen-thawed human semen are significantly correlated with poor post-thaw sperm quality

This study examined the relationship between oxidation–reduction potential (ORP) in frozen-thawed semen and the post-thaw sperm parameters. Levels of ORP were measured in 25 samples from men presenting for routine infertility work-up and were expressed as millivolt (mV)/10⁶ sperm/ml. Frozen-thawed samples were examined for post-thaw total motility (TM%), progressive motility (PM%), total sperm count (TSC) and ORP. The cryo-survival rate (CSR) was calculated as post-thaw TM/pre-freeze TM × 100. Data are provided as median and interquartile range (25th and 75th percentiles). The post-thaw TM% (10.0% [4.00%, 15.1%]), PM% (5.88% [2.97%, 9.33%]) and TSC (12.5 [10.0, 17.5] × 10⁶ sperm) were significantly lower than the pre-freeze TM% (45.9% [32.9%, 59.1%], PM% (31.5% [24.4%, 40.0%] and TSC (120 [90, 250] ×10⁶ sperm) (p < .001). Post-thaw ORP (2.62 [2.52, 3.13] mV/10⁶ sperm/ml) was significantly higher than pre-freeze ORP (0.73 [0.54, 1.21] mV/10⁶ sperm/ml; p < .001). The CSR was 21.7% (11.3%, 31.9%). The post-thaw seminal ORP was negatively correlated with post-thaw TM% (r = −.5; p = .02), PM% (r = −.41; p = .03), TSC (r = −.60; p = .03) and CSR (r = −.52; p = .01). Increased levels of ORP are significantly correlated with poor post-thaw sperm quality and CSR.     

Does brachytherapy of the prostate affect sperm quality and/or fertility in younger men?

OBJECTIVE: Sperm banking prior to surgical procedures which may affect fertility, such as retroperitoneal lymph node dissection, has been well documented. However, such procedures are usually performed in young men. With older men marrying later in life, or remarrying, we wanted to investigate the effects of radiation on prostate cancer patients who wanted to have children afterwards. MATERIAL AND METHODS: We encountered several patients with prostate cancer who decided to undergo brachytherapy and were planning to have more children. We performed a search using PubMed and Ovid for the period 1966-2001 using the key words "fertility", "sperm banking", "radiation effects", "prostate cancer" and "brachytherapy". RESULTS: Of the four young patients we encountered who underwent brachytherapy, we found no significant change in semen parameters post-therapy, and three of them were able to father a child subsequently without any deleterious side-effects. It has been demonstrated in several reports that external-beam radiation therapy is associated with decreased spermatogenesis due to Leydig cell dysfunction and decreased serum testosterone, as well as having a direct effect on spermatogonia. However, there is a scarcity of literature discussing the effects of prostate brachytherapy on spermatogenesis as the patients involved are usually older and usually do not desire to father any more children. As I has a half-life of 60 days, we used an exposure of 10 mR/h at the symphysis pubis and used integration to find the total dose exposed to the testis as follows: Limits 14 400 to 0, S 10e (-In2/1440.Tdt) where T = 14 400 and 20.75 R = 20.75 cGy. Therefore, the total dose was 20.75 cGy x 0.91 = 18.88 cGy. This value is considered too low to have any significant effect on testicular tissues. CONCLUSIONS: We speculate that the effects of prostate brachytherapy on spermatogenesis in prostate cancer patients are minimal. However, due to the half-life of I, we recommend that these patients should wait for at least 3-4 months before trying to conceive. Furthermore, younger men with prostate cancer may want to consider sperm banking prior to brachytherapy if they want to have children in the future.