Immature rat testis sustained long-term development using an integrative model

BACKGROUND: Xenotransplantation has been primarily performed using fresh donor tissue to study testicular development for about 20 years, and whether the cultured tissue would be a suitable donor is unclear. In this study, we combined testicular culture and xenotransplantation into an integrative model and explored whether immature testicular tissue would survive and continue to develop in this model. METHODS: In the new integrative model group, the testes of neonatal rats on postnatal day 8 (PND 8) were cultured for 4 days ex vivo and then were transplanted under the dorsal skin of castrated nude mice. The xenografted testes were resected on the 57th day after xenotransplantation and the testes of rats in the control group were harvested on PND 69. The survival state of testicular tissue was evaluated from morphological and functional perspectives including H&E staining, immunohistochemical staining of 8-OH-dG, immunofluorescence staining, TUNEL assay, ultrastructural study, gene expression and protein analysis. RESULTS: (a) We found that complete spermatogenesis was established in the testes in the new integrative model group. Compared with the control in the same stage, the seminiferous epithelium in some tubules was a bit thinner and there were vacuoles in part of the tubules. Immunofluorescence staining revealed some ACROSIN-positive spermatids were present in seminiferous tubule of xenografted testes. TUNEL detection showed apoptotic cells and most of them were germ cells in the new integrative model group. 8-OH-dG immunohistochemistry showed strongly positive-stained in the seminiferous epithelium after xenotransplantation in comparison with the control group; (b) Compared with the control group, the expressions of FOXA3, DAZL, GFRα1, BOLL, SYCP3, CDC25A, LDHC, CREM and MKI67 in the new integrative model group were significantly elevated (P < 0.05), indicating that the testicular tissue was in an active differentiated and proliferative state; (c) Antioxidant gene detection showed that the expression of Nrf2, Keap1, NQO1 and SOD1 in the new integrative model group was significantly higher than those in the control group (P < 0.05), and DNA methyltransferase gene detection showed that the expression of DNMT3B was significantly elevated as well (P < 0.05). CONCLUSION: The new integrative model could maintain the viability of immature testicular tissue and sustain the long-term survival in vivo with complete spermatogenesis. However, testicular genes expression was altered, vacuolation and thin seminiferous epithelium were still apparent in this model, manifesting that oxidative damage may contribute to the testicular development lesion and it needs further study in order to optimize this model.

Reduced semen quality in patients with testicular cancer seminoma is associated with alterations in the expression of sperm proteins / 亚洲男科学杂志(英文版)

Testicular cancer seminoma is one of the most common types of cancer among men of reproductive age. Patients with this condition usually present reduced semen quality, even before initiating cancer therapy. However, the underlying mechanisms by which testicular cancer seminoma affects male fertility are largely unknown. The aim of this study was to investigate alterations in the sperm proteome of men with seminoma undergoing sperm banking before starting cancer therapy, in comparison to healthy proven fertile men (control group). A routine semen analysis was conducted before cryopreservation of the samples (n = 15 per group). Men with seminoma showed a decrease in sperm motility (P = 0.019), total motile count (P = 0.001), concentration (P = 0.003), and total sperm count (P = 0.001). Quantitative proteomic analysis identified 393 differentially expressed proteins between the study groups. Ten proteins involved in spermatogenesis, sperm function, binding of sperm to the oocyte, and fertilization were selected for validation by western blot. We confirmed the underexpression of heat shock-related 70 kDa protein 2 (P = 0.041), ubiquinol-cytochrome C reductase core protein 2 (P = 0.026), and testis-specific sodium/potassium-transporting ATPase subunit alpha-4 (P = 0.016), as well as the overexpression of angiotensin I converting enzyme (P = 0.005) in the seminoma group. The altered expression levels of these proteins are associated with spermatogenesis dysfunction, reduced sperm kinematics and motility, failure in capacitation and fertilization. The findings of this study may explain the decrease in the fertilizing ability of men with seminoma before starting cancer therapy.

In vitro differentiation of spermatogonial stem cells using testicular cells from Guangxi Bama mini-pig

In this study, we attempted to establish a culture system for in vitro spermatogenesis from spermatogonial stem cells (SSCs) of Bama mini-pig. Dissociated testicular cells from 1-month-old pigs were co-cultured to mimic in vivo spermatogenesis. The testicular cells were seeded in minimum essential medium alpha (α-MEM) supplemented with Knockout serum replacement (KSR). Three-dimensional colonies formed after 10 days of culture. The colonies showed positive staining for SSC-associated markers such as UCHL1, PLZF, THY1, OCT4, Dolichos biflorus agglutinin, and alkaline phosphatase. Induction of SSCs was performed in α-MEM + KSR supplemented with retinoic acid, bone morphogenetic protein 4, activin A, follicle-stimulating hormone, or testosterone. The results showed that STRA8, DMC1, PRM1, and TNP1 were upregulated significantly in the colonies after induction compared to that in testis from 1-month-old pigs, while expression levels of those genes were significantly low compared to those in 2-month-old testis. However, upregulation of ACROSIN was not significant. Replacement of α-MEM and KSR with Iscove's modified Dulbecco's medium and fetal bovine serum did not upregulate expression of these genes significantly. These results indicate that SSCs of Bama mini-pig could undergo differentiation and develop to a post-meiotic stage in α-MEM supplemented with KSR and induction factors.

Value of sperm acrosin activity detection in selecting the method of assisted reproduction for patients with unexplained infertility / 中华男科学杂志

Objective@#To investigate the clinical significance of sperm acrosin activity detection in selecting the method of assisted reproduction for patients with unexplained infertility (UI).@*METHODS@#This retrospective study included 49 UI couples treated by IVFET (49 cycles) after three failures in intrauterine insemination (IUI) and another 95 couples with uterine tube obstruction (UTO) treated by IVF (131 cycles). We analyzed the laboratory data, clinical outcomes and sperm acrosin activity in the two groups of patients. According to the level of sperm acrosin activity of the males, we further divided the UI patients into two subgroups, a 0.05). The sperm acrosin activity was remarkably lower in the UI than in the UTO patients (36.03 vs 61.98 IU/106, P < 0.01), and so was the fertilization rate in the < 36 IU/106 than in the ≥36 IU/106 sperm subgroup (47.7% vs 80.3%, P < 0.01).@*CONCLUSIONS@#The low fertilization rate caused by decreased sperm acrosin activity may be the main cause of infertility and the potential factor of UI. When sperm acrosin activity is < 36 IU/106 sperm, IVF plus shortterm fertilization by remedial ICSI should be preferred to IUI.

Decreases of progressive motility, total motility, and acrosin activity of sperm from oligoasthenoteratospermia males at different time points after sperm activation / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the progressive motility, (PR), total motility (progressive + non-progressive motility, PR + NP), and acrosin activity of sperm from normal and infertile men at different time points after sperm activation.</p><p><b>METHODS</b>Based on the 5th edition of the WHO Laboratory Manual for the Examination and Processing of Human Semen and the results of modified Papanicolaou staining, we divided the semen samples into groups A (normal, n = 28), B (oligoasthenoteratospermia, n = 30), and C (asthenoteratospermia, n = 32). At 1, 24, and 48 hours after sperm activation, we detected sperm PR and PR + NP by CASA and chemical colorimetry, and determined sperm acrosin activity using the modified Kennedy method.</p><p><b>RESULTS</b>Sperm PR and PR + NP were significantly decreased in all the three groups at 1-24 hours and even more significantly at 24-48 hours after sperm activation as compared with the baseline (P < 0.05). Sperm acrosin activity showed remarkable reduction in group A (P = 0. 013) , even more significant at 1-24 hours than at 24-48 hours after sperm activation, but not in groups B and C (P = 0.519 and 0.979).</p><p><b>CONCLUSION</b>Sperm PR, PR + NP, and acrosin activity are all decreased with the extension of time after sperm activation, each in a specific manner. Examination of sperm acrosin activity should be applied as a routine tool in the assessment of male fertility.</p>

Sperm DNA damage and sperm-nucleoprotein transition correlate to acrosin activity and seminal parameters / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the correlation of sperm DNA damage and sperm-nucleoprotein transition with acrosin activity and seminal parameters.</p><p><b>METHODS</b>We collected 535 semen samples, assessed sperm DNA damage by sperm chromatin dispersion test, and analyzed the correlation of sperm DNA damage and sperm-nucleoprotein transition with acrosin activity and seminal parameters according to the WHO criteria.</p><p><b>RESULTS</b>Statistically significant differences were observed in sperm DNA damage among sperm-nucleoprotein transition, acrosin activity, sperm concentration and the percentage of grade a + b sperm (P < 0.01). Sperm DNA damage was positively correlated with age, sperm-nucleoprotein transition, sperm concentration and the percentage of grade d sperm (P < 0.01 or P < 0.05), but negatively correlated with acrosin activity (P < 0.001). Stepwise linear regression analysis demonstrated that age, sperm concentration, the percentage of grade d sperm, sperm-nucleoprotein transition and acrosin activity were independent variables related to the DNA fragmentation index (DFI). The abnormality rates of sperm-nucleoprotein transition, acrosin activity, sperm concentration and graded a + b sperm were significantly higher in the sperm DNA damage group (DFI > or = 30%) than in the normal control (DFI < 30%) (P < 0.01).</p><p><b>CONCLUSION</b>Sperm DNA damage is closely related with sperm-nucleoprotein transition, acrosin activity and seminal parameters, which may become another important independent parameter for the evaluation of sperm quality.</p>

Acrosin release and acrosin activity during incubation in capacitating media using fresh and frozen-thawed dog sperm

We evaluated the effect of time and temperature on acrosin release from the acrosomal cap and the activity of this enzyme during in vitro capacitation in fresh and frozen/thawed dog sperm. Sperm-rich fractions of six ejaculates from three dogs were processed as fresh and frozen samples. Each sperm sample was incubated in canine capacitation medium (CCM) for 0, 1, 2 and 3 h at 20°C and at 37°C. After incubation, the samples were assessed by the indirect immunofluorescent staining technique. The probability of having unlabeled sperm (PUS), indicating acrosin loss, was modelled by a binomial distribution using logistic regression. There was a linear relationship between PUS and time at both temperatures (p<0.001); however, a major percentage of unlabeled sperm was observed in frozen/thawed samples soon after incubation, indicating that the release of acrosin was affected by capacitation time, mainly in frozen samples. Temperature influenced acrosin release only in cryopreserved sperm (p<0.05). Acrosin activity was measured by digestion halos on slides coated with gelatin-substrate film during each time period; a significant increase in the number of large halos was observed in fresh samples throughout the experiment, whereas frozen/thawed sperm showed a decreased rate of halo diameters during culture. Thus, there appears to differences between fresh and frozen dog sperm in terms of acrosin release and the level of acrosin activity in the course of in vitro capacitation.

Protease activity involvement in the passage of mammalian sperm through the zona pellucida

The interaction between acrosome-reacted sperm and zona pellucida proteins is not yet fully understood. Serine protease acrosin and its zymogen proacrosin have been proposed to fulfill this function due to their capacity to bind zona pellucida glycoproteins. However, the molecular mechanism underlying this interaction has been merely speculative. Here we show that fucoidan (a sulfated polysaccharide) and solubilized zona pellucida glycoproteins, but not soybean trypsin inhibitor, are able to detach bound spermatozoa, which suggests that live sperm binds to the zona pellucida in a non-enzymatical way. Interestingly, mild proteolytic digestion with acrosin or trypsin does not modify the structure of the zona pellucida, but rather results in fewer spermatozoa binding to the zona. These results agree with a model where the active site of acrosin digests the zona pellucida and binds through the polysulfate-binding domain through a three-dimensional zona structure rather than a single ligand.

Nandeshi: a powerful inhibitor of human acrosin activity / 中华男科学杂志

<p><b>OBJECTIVE</b>To evaluate the inhibitory effect of Nandeshi, an acrosin inhibitor, on human acrosin activity.</p><p><b>METHODS</b>We collected sperm samples from 10 healthy fertile men and cultured them with Nandeshi at 30 degrees C for 5 minutes at the concentrations of 0. 100, 0.120, 0.144, 0.173, 0.207, 0.249, 0.299, 0.358 and 0.430 mmol/L, with the controls treated with a well-known acrosin inhibitor N-alpha-p-tosyl-L-lysine chloromethylketone (TLCK) at 150.0, 189.8, 213.6, 240.3, 270.3, 304.1 and 342.1 mmol/L. Then we determined the residual activity of human acrosin by improved Kennedy assay.</p><p><b>RESULTS</b>The residual activity of acrosin was negatively correlated with the Nandeshi concentration, and Nandeshi exhibited an inhibition rate about 800 times that of TLCK.</p><p><b>CONCLUSION</b>Nandeshi has a powerful inhibitory effect on human acrosin, and improved Kennedy assay is a simple, practical and highly sensitive technique for the detection of human acrosin activity.</p>

Sperm acrosin activity helps predict IVF-ET outcome / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the effect of sperm acrosin activity on the IVF-ET outcome.</p><p><b>METHODS</b>We analyzed sperm parameters, morphology and acrosin activity for 909 infertile husbands by computer-assisted self-assessment (CASA), modified Papanicolaou staining and N-alpha-benzoyl-DL-arginine-p-nitroanilide (BAPNA), respectively, and detected the rates of fertilization, cleavage, quality embryos, embryo cryopreservation, implantation, clinical pregnancy and abortion. The wives were identified as normal or with mere oviduct problems.</p><p><b>RESULTS</b>The rate of normal sperm morphology and sperm motility, vitality, rapid progressive velocity and concentration were significantly lower in the abnormal acrosin activity group than in the normal one (P < 0.01). Significant positive correlations were observed between acrosin activity and the above-mentioned semen parameters (P < 0.01). There were no significant differences in the number of retrieved eggs, the rates of cleavage, quality embryos, embryo cryopreservation, non-embryo transfer cycles and miscarriages, and the number of transferred embryos between the two groups (P > 0.05). The fertilization rate, the percentage of transfer cycles with only 1 embryo and the rate of implantation and clinical pregnancy were notably higher in the normal acrosin activity group than in the abnormal one (P < 0.01).</p><p><b>CONCLUSION</b>Sperm acrosin activity is closely related with semen parameters, and it helps to predict the sperm fertilizing capacity and IVF-ET outcome.</p>

Effect of some antioxidants on acrosin amidase activity of chilled stallion spermatozoa

The effect of some antioxidants on acrosin amidase activity of chilled stallion spermatozoa was investigated in this study. Nine ejaculates were collected from six Arabian horses. A comparison between four antioxidants namely, sodium pyruvate [0.5 mg/ml], sodium thiosulfate [STS, 1.0 mg/ml], bovine serum albumin [BSA, 5.0 mg/ ml], zinc chloride [0.15 mg/ml] and a mixture of them was studied in a chemically-defined stallion semen extender [Tris-egg yolk] at 5°C. The comparison was based on sperm viability, acrosin amidase activity and changes in the levels of extra-cellular alanine aminotransferase [ALT]. Results of the present experiments revealed that sodium pyruvate and the mixture of antioxidants were most effective for improving viability and acrosin amidase activity of stallion spermatozoa. Low values of ALT in the extracellular medium were coincided with high values of acrosin amidase activity of equine stallion spermatozoa during storage at 5°C

Assessment of released acrosin activity as a measurement of the sperm acrosome reaction / 亚洲男科学杂志(英文版)

<p><b>AIM</b>To develop a method for assessing sperm function by measuring released acrosin activity during the acrosome reaction (AR).</p><p><b>METHODS</b>Human semen samples were obtained from 24 healthy donors with proven fertility after 3-7 days of sexual abstinence. After collection, samples were liquefied for 30 min at room temperature. Standard semen parameters were evaluated according to World Health Organization (WHO) criteria. Calcium ionophore A23187 and progesterone (P4) were used to stimulate the sperm to undergo AR. After treatment, sperm were incubated with the supravital dye Hoechst33258, fixed in a glutaraldehyde-phosphate-buffered saline solution, and the acrosomal status was determined by fluorescence microscopy with fluorescein isothiocyanate-labeled Pisum sativum agglutinin (FITC-PSA). The percentage of sperm undergoing AR (AR%) was compared to sperm acrosin activities as assessed by spectrocolorimetry. The correlation between AR% and acrosin activity was determined by statistical analysis.</p><p><b>RESULTS</b>The AR% and released acrosin activity were both markedly increased with A23187 and P4 stimulation. Sperm motility and viability were significantly higher after stimulation with P4 versus stimulation with A23187 (P < 0.001). There was a significant positive correlation between released acrosin activity and AR% determined by FITC-PSA staining (r=0.916, P < 0.001).</p><p><b>CONCLUSION</b>Spectrocolorimetric measurement of released acrosin activity might serve as a reasonable alternative method to evaluate AR.</p>

Reproductive toxic effects in the testes of BALB/c mice exposed to total diesel exhaust

This study used 50 male BALB/c mice divided into three groups [20 in each of test groups A and B and 10 in the control group C]. Group A and group B were exposed to total diesel exhaust [TDE]. The TDE exposures were performed in a cubic wooden box chamber [side length 50 cm] filled with TDE from a diesel fueled car. Group A was moved to the diesel filled chamber and exposed once a day for 30 minutes and group B was moved to the filled chamber and exposed once a day for 60 minutes. The control group was similarly manipulated for 60 minutes without filling the chamber with TDE. The experiment was carried out six days a week for 120 days. Four animals from group A and six animals from group B did not survive to the end of the experiment while the control animals did not have mortalities. Five of the remaining mice from each test group and 2 controls were sacrificed on the 40th day and on the 80th day. Remaining six mice in group A and four mice in group B and six mice from group C were sacrificed at the end of the experiment [120th day]. Testes and vasa efferentia were removed, testes were prepared into sections for histological, immunohistochemical staining and testicular biopsies [5 mg each] were used for Western blotting experiments to detect acrosomal proteins. Vas spermatozoa were prepared as smears for immuno-histochemical study. Down regulation of spermatogenesis reflecting structural damage of the seminiferous tubules was observed in animals sacrificed on the 40th day, this was progressive with time of exposure as seen in samples obtained on the 80th and 120th days The dependence on exposure time was also clear from comparison of sections from groups A and B, Severe oligozoospermia was detected in group A by the end of 80th days and in group B by the end of the 40th day. By the end of the experiment [120 days], the seminiferous tubules from the testes of the two test groups A and B were containing Sertoli cells only. Immunohistochemical staining of testicular sections and vas sperm suspensions using monoclonal antibodies for internal acrosomal proteins revealed a concomitant ultrastructural damage of spermatozoa in the form of defective or absent acrosome and increased proportion of abnormal sperm head morphology. The progressive decrease of sperm and spermatid -specific proteins in testicular biopsies was observed in the immuno blots. It is concluded that exposure to diesel exhaust has a massive reproductive toxicity in male mice manifested by suppression of spermatogenesis and abnormal ultra structures of vas spermatozoa. Also, the reproductive toxic effect of diesel exhaust exposure is both dose [exposure time]-dependent and duration [repeated exposures]-dependent

Progress in the study of immunocontraceptive antigens / 中华男科学杂志

Fertility management is a global issue of medical, economic, and social consequence. Although many methods have been devised to inhibit reproduction, more acceptable alternatives are still needed. Regulation by immune intervention is a promising technology as applied to human beings. The objective of this review is to indicate several immunocontraceptive antigens.

Study of the changes of acrosomal enzyme and nitric oxide synthase, superoxide dismutase of infertile patients with positive antisperm antibody in seminal plasma / 中华男科学杂志

<p><b>OBJECTIVE</b>To study the impacts of positive antisperm antibody (AsAb) in seminal plasma on acrosomal enzyme activity, nitric oxide synthase (NOS) and superoxide dismutase (SOD) levels of spermatozoa.</p><p><b>METHODS</b>Swatch from 40 infertile patients with positive AsAb in seminal plasma as experimental group, and 40 fertile men as control group. Acrosomal enzyme activity was detected by the BAEE/ADH unitive method, NOS was detected by the redoxreaction assay, and SOD level was measured xanthine oxidase method.</p><p><b>RESULTS</b>Compared with control group, acrosomal enzyme activity of spermatozoa of experimental group was significantly decreased (P <0.01), NOS activity was apparently increased (P < 0.01), and SOD level in seminal plasma was markedly decreased (P<0.01).</p><p><b>CONCLUSION</b>It may be possible that the positive AsAb in seminal plasma beget infertility through the changes of acrosomal enzyme of spermatozoa, SOD and NOS activities in seminal plasma.</p>

Analysis of relationship between semen quality and sperm acrosin activity / 中华男科学杂志

<p><b>OBJECTIVE</b>To analyze the activity of human sperm acrosin and semen parameters in male infertile patients and discuss the effect of sperm acrosin activity on semen quality.</p><p><b>METHODS</b>Activity of human sperm acrosin, PMN-elastase, fructose, alpha-glucosidase, zinc, acid phosphatase levels and HOST of 214 male infertile patients were detected. Semen analysis was performed using WLJY-9000 WeiLi colorful semen quality analyzer. There were 111 cases of normal activity of human sperm acrosin (48.2 - 218.7 microIU/10(6) sperm), 103 cases abnormal (< 48.2 microIU/10(6) sperm). Using the group of normal activity of sperm acrosin to be the control, semen parameters was analyzed and compared with those of the group of abnormal activity of sperm acrosin.</p><p><b>RESULTS</b>There were significant difference (P < 0.001) between the 2 groups (normal and abnormal) in the areas of sperm density, motile sperm rate, percentage of grade (a + b) sperm and HOST. There were also significant difference in PMN-elastase, fructose and alpha-glucosidase (P < 0.05). There was no difference among sperm volume, zinc and acid phosphatase (P > 0.05).</p><p><b>CONCLUSION</b>There was a strong correlation between the activity of human sperm sperm acrosin and semen quality. Activity of sperm acrosin is a reliable index of semen quality.</p>

Mechanism of IL-6 on acrosome reaction in human sperm / 中华男科学杂志

<p><b>OBJECTIVE</b>To explore the mechanism of interleukin-6 (IL-6) on acrosome reaction (AR) in human sperm.</p><p><b>METHODS</b>Sperm acrosin activity was measured by BAEE/ADH and AR evaluated by FITC-PSA.</p><p><b>RESULTS</b>IL-6 could induce the activity of acrosin and superoxide dismutase (SOD) and enhance AR in human sperm. AR was not induced by extracellular Ca2+, and IL-6-induced AR did not occur in Ca2(+)-free medium. Calphostin C, one inhibitor of the protein kinase C (PKC), could block IL-6-induced AR in human sperm.</p><p><b>CONCLUSION</b>IL-6 could induce AR by stimulating the activity of acrosin and SOD in human sperm, which also involves the activation of PKC, and requires the presence of extracellular Ca2+.</p>

Etiologic analysis of globozoospermia: a 1-case report / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the etiologic factors of globozoospermia.</p><p><b>METHODS</b>Routine semen analysis, sperm DNA special staining and chromosomal karyotype detection of peripherical blood lymphocytes were performed in a globozoospermia patient.</p><p><b>RESULTS</b>Round-headed spermatozoa were lack of acrosome and the acrosin activity was low. Meanwhile, there was an additional band located in the Y chromosomal short arm.</p><p><b>CONCLUSION</b>Lack of acrosome, low acrosin activity and abnormality of chromosome may be the main reasons for globozoospermia.</p>

Proacrosin-acrosin activity in capacitated and acrosome reacted sperm from cryopreserved bovine semen

Acrosin activity is associated with normal fertility in human and bovine spermatozoa. The aim of the study was to determine the variation of the enzyme activity in the proacrosin-acrosin system in capacitated and acrosome recated cryopreserved bovine sperm. Enzyme activity was assessed spectrophotometrically using N-alpha-benzoyl-DL-arginine p-nitroanilide (BAPNA) as specific substrate for acrosin at pH 8. Capacitation with heparin and quercitin failed to induce conversion of proacrosin to acrosin. An increase in acrosin activity produced by the presence of progesterone, in a dose-dependent manner, was related with the induction of true acrosome reaction. The total level of acrosin activity registered showed that 96 per cent of acrosin of capacitated sperm samples and control is present in the zymogen form. Moreover, progesterone is capable of duplicating the level of active enzyme, indicating that enzyme activity changes are related to acrosome reaction, suggesting that only a minor proportion of the total of proacrosin-acrosin system is required in the exocytotic process on cryopreserved bovine sperm.

Relationship between acrosin activity of human spermatozoa and oxidative stress / 亚洲男科学杂志(英文版)

<p><b>AIM</b>To study the association between seminal oxidative stress and human sperm acrosin activity.</p><p><b>METHODS</b>It is a prospective study consisting of 30 infertile men and 12 fertile normozoospermic volunteers. A full history, clinical examination and scrotal ultrasound were done to exclude other related factors such as smoking and varicocele. Presence of white blood cells (WBCs) in semen samples was evaluated by peroxidase staining. Lipid peroxidation in spermatozoa was induced after incubating with ferrous sulphate (4 mmol/L) and sodium ascorbate (20 mmol/L). Induced peroxidation of spermatozoa was assessed by determining the production of thiobarbituric acid reactive substances (TBARS). Acrosin activity was measured using the gelatinolysis technique. The halo diameters around the sperm heads and the percentages of spermatozoa showing halo formation were evaluated. An acrosin activity index was calculated by multiplying the halo diameter by the halo formation rate.</p><p><b>RESULTS</b>A significant difference was observed in acrosin activity parameters and TBARS levels between samples with WBCs (1 multiply 10(6)/mL of ejaculate) and those without. This difference was also noted between the normozoospermic and the oligoasthenoteratozoospermic semen samples. The TBARS production by spermatozoa had a significant negative correlation with the acrosin activity index (r = -0.89, P 0.001).</p><p><b>CONCLUSION</b>The presence of oxidative stress in an individual with leukocytospermia and/or abnormal semen parameters is associated with impaired sperm function as measured by its acrosin activity.</p>