Copper: a biphasic regulator of caprine sperm forward progression

Abstract

Copper is essential for spermatogenesis and its presence has been demonstrated in male and female reproductive fluids in several mammalian species. However, little is known about the physiological significance of this trace element in the regulation of forward progression of mammalian sperm cells which is essential for sperm fertility potential in vivo. The purpose of this investigation was to determine the physiological role of the bivalent copper ion (Cu²⁺) on mammalian sperm forward motility using a chemically-defined medium and caprine cauda epididymal sperm model. Sperm forward motility was significantly enhanced by Cu²⁺ in a dose-dependent manner; maximal activation (approx 20%) was noted at the 5?µM level of the metal. Above 10?µM Cu²⁺ sperm motility decreased, showing that Cu²⁺ exerts a biphasic regulation on sperm motility. These findings have been confirmed using a spectrophotometric motility assay, an objective method of motility analysis. At lower concentrations (up to 5?µM), copper enhanced sperm membrane lipid peroxidation as well as the level of intra-sperm cyclic adenosine mono phosphate (c-AMP), but at a higher level it caused marked inhibition of both of the biochemical parameters. The observed correlation of Cu²⁺-dependent biphasic modulation of sperm membrane lipid peroxidation and intrasperm c-AMP with sperm forward motility is consistent with the view that Cu²⁺ regulation of sperm motility is mediated by membrane lipid peroxidation, which in turn modulates the level of intra-sperm c-AMP, a well-known activator of sperm motility.     

Xenoestrogens at picomolar to nanomolar concentrations trigger membrane estrogen receptor-alpha-mediated Ca2+ fluxes and prolactin release in GH3/B6 pituitary tumor cells

Xenoestrogens (XEs) are widespread in our environment and are known to have deleterious effects in animal (and perhaps human) populations. Acting as inappropriate estrogens, XEs are thought to interfere with endogenous estrogens such as estradiol (E2) to disrupt normal estrogenic signaling. We investigated the effects of E2 versus several XEs representing organochlorine pesticides (dieldrin, endosulfan, o',p'-dichlorodiphenylethylene), plastics manufacturing by-products/detergents (nonylphenol, bisphenol A), a phytoestrogen (coumestrol), and a synthetic estrogen (diethylstilbestrol) on the pituitary tumor cell subline GH3/B6/F10, previously selected for expression of high levels of membrane estrogen receptor-alpha. Picomolar to nanomolar concentrations of both E2 and XEs caused intracellular Ca2+ changes within 30 sec of administration. Each XE produced a unique temporal pattern of Ca2+ elevation. Removing Ca2+ from the extracellular solution abolished both spontaneous and XE-induced intracellular Ca2+ changes, as did 10 microM nifedipine. This suggests that XEs mediate their actions via voltage-dependent L-type Ca2+ channels in the plasma membrane. None of the Ca2+ fluxes came from intracellular Ca2+ stores. E2 and each XE also caused unique time- and concentration-dependent patterns of prolactin (PRL) secretion that were largely complete within 3 min of administration. PRL secretion was also blocked by nifedipine, demonstrating a correlation between Ca2+ influx and PRL secretion. These data indicate that at very low concentrations, XEs mediate membrane-initiated intracellular CCa2+ increases resulting in PRL secretion via a mechanism similar to that for E2, but with distinct patterns and potencies that could explain their abilities to disrupt endocrine functions.     

SPERM-IMMOBILIZING ANTIBODIES BLOCK CAPACITATION IN HUMAN SPERMATOZOA

Sperm-immobilizing antibodies block human fertilization by interfering with the acrosome reaction (AR). To clarify the mechanism of blockage of AR by sperm-immobilizing antibodies, the authors examined their effects on the increase of intracellular free Ca 2+ concentration induced by follicular fluids (Ca 2+ influx) in spermatozoa and on their capacitation. Sperm-immobilizing antibodies did not suppress Ca 2+ influx induced by follicular fluid, but they inhibited capacitation of human spermatozoa. Namely &#106 %AR (%AR after addition of an AR inducer - %AR before treatment) induced by progesterone was significantly ( p <. 0001) lower when spermatozoa were incubated in human tubal fluid medium cotaining antibody-positive serum (1.2%), compared to that when incubated in control medium (19.2%). Furthermore, the proportion of both spermatozoa that became capacitated and ones that had become capacitated decreased significantly ( p <. 0001) after 2, 4, and 6 h of incubation in medium containing antisperm antibody-positive serum, compared to those of spermatozoa incubated in control medium. In conclusion, sperm-immobilizing antibodies may be closely related to their blockage of capacitation.     

POTASSIUM INCREASES INTRACELLULAR CALCIUM SIMULATING PROGESTERONE ACTION IN HUMAN SPERM

Progesterone (P) and zona pellucida are known to induce acrosome reaction in human sperm by increasing cytosolic calcium. High concentrations of potassium ions (K⁺) improve the rate of acrosome reaction in human sperm in vitro. This article determined whether the effect of K⁺ on the acrosome in human sperm is mediated by increasing intracellular calcium ([Ca²⁺]i). The effect of K⁺ on [Ca²⁺]i was examined by using Fura 2 as the fluorescent indicator. The effect of K⁺ and P on [Ca²⁺]i in sperm and the involvement of ion channels was compared. Motile sperm were collected by the swim-up method from semen of healthy volunteers and capacitated overnight in BWW containing 0.5% BSA. Incubation of capacitated sperm with different concentrations of potassium chloride (1.25-20 mM) resulted in dose-dependent increase in [Ca²⁺]i similar to that observed with P. The increase in [Ca²⁺]i by K⁺ and P was blocked by the addition of EGTA, a Ca²⁺ chelator. K+-induced change in [Ca²⁺] was not altered by the addition of dihydropyridine derivatives. The combined treatment of K+ (20 mM) and P (0.75 mug/mL) caused an additive effect on the increase in [Ca²⁺]i. It would appear that human sperm plasma membrane possess different Ca2+ channels responsive to P and K⁺.     

USEFULNESS OF SPERM QUALITY ANALYZER-V (SQA-V) FOR THE ASSESSMENT OF SPERM QUALITY IN INFERTILE MEN

The aim of this study was to evaluate the correlation of new of Sperm Quality Analyzer (SQA-V) with the computer-assisted sperm analysis (CASA) and manual semen analysis estimates. One hundred five fresh semen samples were analyzed using SQA-V and CASA and manual semen analysis. Significant correlations of sperm concentration (p < 0.0001), sperm motility (p < 0.0001), and normal morphology (p < 0.0001) were observed between SQA-V variables and manual semen analysis estimates. There also were significant correlations of sperm concentration (p < 0.0001), sperm motility (p < 0.0001), and sperm velocity (p = 0.0235) between SQA-V variables and CASA estimates. Meanwhile, it did not correlate with amplitude of lateral head displacement, beat cross frequency, lineality assessed by CASA. The value of the sperm concentration and the sperm motility measured by SQA-V showed high correlations with the value of those measured by CASA and manual semen analysis. In addition, velocity and sperm morphology may also be evaluated to some extent using SQA-V.     

Fucoidin inhibits the zona pellucida-induced acrosome reaction in human spermatozoa

We recently reported that fucoidin (a polymer of predominantly sulfated L-fucose) significantly inhibits tight binding of human sperm to the human zona pellucida in vitro and that several oligosaccharides obtained after acid hydrolysis possess sperm-zona pellucida binding inhibitory activity equal to the original fucoidin. This inhibition may be specific to sperm-zona interactions or may be the consequence of the interruption of capacitation, a series of biochemical and physiological events leading to final sperm maturation, that must occur for successful fertilization. Completion of capacitation is most often determined by assessing two end-points of the process: acquisition of hyperactivated motility and ability to complete the acrosome reaction. Here, we examined the effects of fucoidin on these two end-points of capacitation in vitro. Fucoidin did not affect the proportion of sperm with hyperstimulated motility. Neither did fucoidin cause an increase in sperm that had spontaneously acrosome-reacted at 4.5 hours compared to controls as evaluated by indirect immunofluorescence using the acrosomal marker, monoclonal antibody, T-6. Comparable percentages of sperm had completed the acrosome reaction when exogenously stimulated by calcium ionophore A23187 with and without the addition of fucoidin. However, in the presence of fucoidin, stimulation of the acrosome reaction by acid solubilized human zonae pellucidae was significantly inhibited. These data indicate that fucoidin does not impede the normal progression of capacitation. These results provide strong evidence to support the hypothesis is that the inhibitory effect of fucoidin is at the level of the sperm membrane since inhibition can be bypassed by increasing intracellular calcium directly with a calcium ionophore.     

The inhibitory effect of some ionophores on human sperm motility

The inhibitory effect of five ionophores, namely, A23187, nonactin, nigericin, monensin and m-chlorocarbonyl cyanide phenylhydrazone, on human sperm motility were measured with a trans-membrane migration method. The concentrations of A23187 and nigericin that decreased sperm motility to 50% of control were 20 microM (10.5 mg/l) and 8 microM (5.8 mg/l), respectively. Because these two ionophores were more potent than previously reported membrane-active sperm-immobilizing agents, we propose that ionophores could be a new category of vaginal contraceptive if a pharmaceutical preparation that is safe to be administered in vivo can be developed.     

Increase of Intracellular free [CA2+] in Single Human Motile Spermatozoa Treated with Human Follicular Fluid

The intracellular free [Ca²⁺] concentration ([Ca²⁺]_i) in individual human sperm was measured using a fluorescent Ca²⁺ indicator. In 18 of 23 motile sperm (78.3%), [Ca²⁺]_i increased significantly and promptly after addition of 20% human follicular fluid (hFF), but in the others it did not increase. The mean resting [Ca²⁺]_i level of sperm in which [Ca²⁺]_i increased after addition of 20% hFF (the influx group) was significantly lower than those in which it did not increase (112.8 ± 40.1 nM vs. 156.9 ± 13.5 nM,p <. 05). After addition of 20% hFF, the mean [Ca²⁺]_i in the influx group reached a peak value of 210.7± 24.7 nM within 30 s and then decreased slowly; the mean [Ca²⁺]_i values 1, 5, 10, and 15 min after addition of 20% hFF were 179.3 ± 31.4, 174.3 ± 30.2, 172.5 ± 27.8, and 175.1 ± 27.2 nM, and all values were significantly higher than the resting level (p <. 01). The frequency distribution of [Ca²⁺]_i after addition of 20% hFF was shifted toward higher concentrations (p <. 01). However, the addition of 20% hFF did not increase the percentage of live acrosome reaction (before 3.8 ± 0.9% vs. after 2.9 ± 0.5%, respectively). Thus, hFF increased [Ca²⁺]_i in about 80% of the motile sperm. Relatively high [Ca²⁺]_i levels persisted for at least 10–15 min after its addition. However, hFF did not trigger a rapid response in acrosome reaction.     

Validation of an amphibian sperm inhibition toxicological test method using zinc

Analysis of sperm has been investigated as a possible method to examine the toxicity of environmental contaminants. The amphibian sperm inhibition toxicological test (ASITT) method examines the effects of contaminants on Xenopus laevis (African clawed frog) sperm motility and path trajectories. As part of a preliminary validation of the method, the effects of increasing divalent metal ion, zinc (Zn2+), on X. laevis sperm motility were examined. We hypothesized that Zn2+ concentration would have significant inhibitory effects on percent sperm motility, velocities, and trajectories. The Zn2+ was added to a control solution in concentrations from 0 to 1,417 microg/L. Sperm cells were videotaped at 30 frames per second under x 400 microscope, and percent motility was recorded and paths were mapped by marking the change in position of the sperm head over a period of 1 s. Sperm motility was categorized as progressive, hyperactivated, idle, or nonmotile, and velocities and trajectories were calculated on the basis of x,y coordinates. Increasing Zn2+ concentrations caused a significant exponential decay in percent total motility and progressive motility. Straight-line velocity increased with increasing Zn2+ concentrations. Overall, results suggest that Zn2+ may be interfering with cellular processes, such as cellular respiration, flagellar bending, or ion exchange, thereby inhibiting sperm motility.     

FERTILIZATION RATES OF SMALL-HEAD SPERM IN CONVENTIONAL IVF AND ICSI

Five men produced 23 ejaculates in 23 assisted reproductive technology cycles for semen analysis. In 11 of the 14 ICSI cycles and in 5 of the 9 IVF cycles, small-head sperm were found in more than 70% of the cells, having a length of <3.5 µm and a width of <2 µm. In 6 of the 14 ICSI cycles, the embryologist who performed the ICSI was not alerted to the presence of small-head sperm. Subsequent fertilization rate was significantly lower than a) the fertilization rate of the remaining 5 ICSI cycles that acted as a control, in which the embryologist was alerted to the presence of small-head sperm, and b) was lower than the 3 ICSI cycles with normal sperm head ( p <. 05). The 8 cell embryo and blastocyst rates in the former group were also decreased, although not significantly, compared to the latter groups. Likewise, in the 5 IVF cycles with small-head sperm, the fertilization rate was significantly lower than in the 4 IVF with normal sperm head ( p <. 05). Embryologists should be alerted when a high percentage of small-head sperm are detected during routine semen analysis because they may be associated with reduced fertilization and embryo outcome.     

INFLUENCE OF ACETAZOLAMIDE ON AQP1 GENE EXPRESSION IN TESTIS AND ON SPERM COUNT/MOTILITY IN EPIDIDYMIS OF RATS

Acetazolamide (Ace) is a putative inhibitor of carbonic anhydrase (CA), an enzyme that catalyzes the equilibration of carbon dioxide and carbonic acid and plays a key role in HCO 3 - and water reabsorption and acid secretion. Aquaporins (AQPs) are channel-forming membrane glycoproteins that mediate water reabsorption by the renal tubules and other organs of mammals. AQP1 and CAII or CAIV share many common biological properties. Previous studies have shown that AQP1 and CA are located at the same sites in cells of the male reproductive tract. In the present study, Ace at a dose of 40 mg/kg/d &#50 14, administered per os, suppressed AQP1 gene expression and inhibited CA activity in rat testis. On day 7 of treatment the epididymal sperm motility was significantly reduced, while on day 14 a decrease in sperm count occurred. Ace caused a marked downregulation of AQP1 gene expression; significant suppression occurred on days 7 and 14. Moreover, CA activity was totally blocked throughout the treatment period. The present findings suggest that the reduction of rat sperm motility and count by Ace can be attributed to its capacity to downregulate AQP1 water channel gene expression.     

Effects of mechanical stresses on sperm function and fertilization rate in mice

In this study, we investigated whether any of the observed changes in mouse sperm function tests secondary to mechanical stresses (centrifugation and pipetting) correlate with sperm fertilization ability. Chinese Kunming mice were used as sperm and oocyte donors. Sperm samples were allocated evenly into centrifugation, pipette, and control groups. Sperm plasma membrane integrity (PMI), mitochondrial membrane permeability (MMP), baseline and stimulated intracellular ROS, and sperm fertilization ability were measured by hypo-osmotic swelling, flow cytometry, and fertilization tests. Parallel studies were conducted and all tests were repeated six times. Our results showed that after centrifugation, the progressive motility, average path velocity, and overall sperm motility and PMI decreased significantly (p < 0.05). In addition, the MMP level decreased significantly in viable sperm when the centrifugation condition reached 1,400 g × 15 minutes (p < 0.05). When pipetting was performed two or more times, progressive motility, average path velocity, and overall sperm motility decreased significantly (p < 0.05); when it was performed four or more times, sperm membrane integrity and intracellular basal ROS level of viable sperm was also significantly decreased (p < 0.05). In conclusion, various mechanical stresses seem to affect sperm function, however this does not appear to alter fertilization rate. Laboratory handling steps should be minimized to avoid unnecessary mechanical stresses being applied to sperm samples.     

Effects of Genistein Isoflavone (4',5',7-Trihydroxyisoflavone) and Dexamethasone on Functional Characteristics of Spermatozoa

Caudal epididymal spermatozoa were used to study the influence of genistein isoflavone and dexamethasone (dxm) on the functional characteristics of spermatozoa. The effects of genistein alone and in combination with dxm on sperm motility, sperm morphology, spontaneous acrosome reaction (AcR), and ionophore A23187-induced AcR were investigated. The FITC-PSA/Hoechst 33258 staining procedure was used to assess sperm cell viability and AcR status and thus to differentiate between true AcR and acrosome degeneration. The overall results indicated that (1) lower doses of genistein alone, or in combination with dxm, did not significantly influence sperm motility or sperm morphology; (2) ionophore A23187 induced AcR in rat spermatozoa; (3) there appeared to be no direct correlation between sperm motility and AcR, (4) higher doses of genistein, alone or in combination with dxm, significantly interfered with percentage sperm motility and caused significant detachment of sperm heads but did not cause morphological defects; and (5) higher doses of genistein caused significant decrease in sperm acrosome reactivity with long duration of exposure. In view of the fact that sperm capacitation and AcR are physiological prerequisites for successful fertilization of oocytes, the findings suggest that chronic exposure of spermatozoa to high doses of genistein could be associated with infertility problems through suppression/inhibition of AcR and sperm motility. Dexamethasone did not appear to influence the effect of genistein on the functionality of postspermatogenic spermatozoa.     

EFFECTS OF SEMEN CHARACTERISTICS ON IUI COMBINED WITH MILD OVARIAN STIMULATION

To determine the influence of sperm parameters inseminated on the outcome of intrauterine insemination (IUI) in patients undergoing ovarian stimulation with clomiphen citrat (CC) or human menopausal gonadotropin (HMG) therapy, a retrospective review was performed for 2 years on data from the IUI program. 190 couples underwent a total of 268 IUI cycles in which CC or HMG was used for ovulation induction. The initial sperm concentration (mil/ml), motility (percent), preprocessing total motile sperm (TMS) count (million), fast motile sperm (percent) and postprocessing sperm concentration (mil/ml), motility (percent), TMS count, fast motile sperm (percent), sperm morphology, hypoosmotic swelling (HOS) scores, semen leuocytes, and bacteria were analyzed. 268 inseminations were followed by a pregnancy rate of 12% and couple pregnancy rate of 17%. On multivariable logistic regression analysis, total motile sperm (TMS) count, percent motility, and percent of fast motile sperm were independent prognostic factors of fertility. The impact of the preprocessing and postprocessing sperm parameters on pregnancy outcome after IUI was evaluated. There was a trend toward an increasing percent of conception with increasing TMS count, motility, and percent of fast motile sperm. The TMS count, motility and percent of fast motile sperm independently predict success with IUI. Patients with original sperm motility ≥30% had a higher cumulative pregnancy rate (74%) than patient with motility <30% (p < 0.005). Pregnancy rate increased 4 times with motility of ≥30%.     

CHANGES IN SEMEN QUALITY IN JERUSALEM BETWEEN 1990 AND 2000: A CROSS-SECTIONAL AND LONGITUDINAL STUDY

The aim of the present study was to investigate whether sperm quality has changed during the years 1990-1999 among men residing in Jerusalem, Israel, who were involved in treatment by intrauterine insemination (IUI). Both cross-sectional and longitudinal analyses were performed. A total of 2638 male partners in couples that underwent treatment by IUI participated in the cross-sectional investigation. Of them, 417 men (16%) were included in the longitudinal study. Total sperm counts and percent motility were evaluated on an annual basis to assess changes over 10 years from 1990 through 1999. A significant downward trend in sperm count and motility was demonstrated in the cross-sectional study. Sperm count decreased by 5.2×10 6 ±0.9×10 6 ( p <. 0001) each year and percent motility declined by 0.50±0.14% ( p =.0003). Similar changes were found in the longitudinal evaluation, but they were not statistically significant. These data suggest that during the last decade in Jerusalem, sperm count and motility declined significantly among men involved in infertile relationships and treated by IUI.     

EFFECT OF SEMINAL PLASMA CALCITONIN LEVELS ON SPERM MOTILITY

This study investigated the effect of the seminal and blood plasma calcitonin levels on the sperm motility in idiopathic infertile patients. The number of sperm cells and their motility were evaluated in the spermiograms of 52 idiopathic infertile patients. The levels of seminal plasma calcitonin were studied with double antibody technique using a DPC kit. Fifty-two patients were divided into 2 groups according to the motility rates of sperm and 20 healthy volunteers were assigned to a control group. The difference between the groups was evaluated by using Kruskall-Wallis and Mann-Whitney U tests, and the correlation of seminal and blood calcitonin levels with sperm motility were determined. The difference in motility rates between the 3 groups was statistically significant ( p =. 000, p <. 05). Blood plasma calcitonin levels were in normal ranges in all cases and no significant difference was found among the 3 groups ( &#104 2 = 2.7219, p =. 2589, p >. 05). While sperm motility was correlated with seminal calcitonin levels ( r =. 8581), blood calcitonin levels did not show a correlation with sperm motility rate ( r = -.0265). Moreover, there was no correlation between seminal and blood plasma levels of calcitonin ( r = -.0010). Motility rates decreased in the patients with low seminal calcitonin levels and seminal calcitonin levels had a significant effect on sperm motility.     

Effect of density gradient centrifugation on reactive oxygen species in human semen

Density gradient centrifugation can separate motile sperm from immotile sperm and other cells for assisted reproduction, but may also remove antioxidants from seminal plasma, resulting in oxidative stress. Therefore, we investigated reactive oxygen species (ROS) concentrations and distribution in semen before and after density gradient centrifugation. We assessed semen volume, sperm concentration, sperm motility, and ROS levels before and after density gradient centrifugation (300 x g for 20 minutes) in 143 semen samples from 118 patients. The ROS removal rate was evaluated in ROS-positive samples and ROS formation rate in ROS-negative samples. Thirty-eight of 143 untreated samples (26.6%) were ROS-positive; sperm motility was significantly lower in these samples than in ROS-negative samples (p < 0.05). After density gradient centrifugation, only seven of the 38 ROS-positive samples (18.42%) exhibited a ROS-positive lower layer (containing motile sperm) with a ROS removal rate of 81.58%, whereas the upper layer was ROS-positive in 24 samples (63.16%). In the ROS-negative group (n = 105), ROS was detected in 19 samples after centrifugation (18.10%, ROS generation rate), of which 18 were ROS-positive only in the upper layer or interface and the other was ROS-positive in both layers. Density gradient centrifugation can separate motile sperm from immotile sperm as well as remove ROS (including newly generated ROS). This data supports the view that density gradient centrifugation can select motile spermatozoa without enhancing oxidative stress.

Abbreviations: ROS: reactive oxygen species; SOD: superoxide dismutase; GPx: glutathione peroxidase; DNA: deoxyribonucleic acid; DGC: density gradient centrifugation; IUI: intrauterine insemination; IVF: in vitro fertilization; HEPES: 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid; EDTA: ethylenediaminetetraacetic acid; HTF: HEPES-buffered human tubal fluid; IMSI: intracytoplasmic morphologically selected sperm injection; SMAS: sperm motility analyzing system; CASA: computer-assisted semen analyzer; WHO: World Health Organization     

Inhibition of the metabolism and motility of human spermatozoa by various alkaloids

The ability of several alkaloids to inhibit the metabolism and motility of human spermatozoa has been investigated.Of the agents tested, chloroquine was the most effective in inhibiting sperm metabolism (production of carbon dioxide and lactic acid) and motility. It was active at a concentration of 3.6 × 10^?5M. Quinine and Quinacrine were active at concentrations of 5 × 10^?4M and emetine required concentrations as high as 3.6 × 10^?3M to achieve an inhibitory effect. Detailed studies with emetine showed that the time needed for inhibition of sperm motility was inversely proportional to the drug concentration and directly related to the sperm density. In addition, the inhibition was shown not to be reversible.     

ASTHENOZOOSPERMIA: ANALYSIS OF A LARGE POPULATION

Spermatozoa travel a long distance to meet and fertilize the oocyte, so sperm motility is a requisite for normal fertilization. Asthenozoospermia, or low sperm motility, is a common cause of human male infertility. This is a retrospective study (1992-1999) to document the prevalence of this pathology in infertile men and to clarify the probable factors associated to its etiology. The prevalence was 18.71% for asthenozoospermia and 63.13% for asthenozoospermia associated with oligo- and/or teratozoo-spermia; thus, 81.84% of the studied samples had altered motility. Leukocytospermia, the ratio of germ cells/sperm, anti-sperm antibodies, consistency, biochemical markers of accessory sex glands, and sperm response after swim-up were studied in normospermic (N), asthenozoospermic (A), and combined asthenozoospermic (C) samples. No significant difference was found in the frequency of leukocytospermia among groups. The rate of germ cells/(spermatozoa + germ cells) between C and N ( p <.01), and C and A ( p <.01) was statistically different, while no difference was found on comparing N and A. MAR-test over 40% was found in 6% of the A samples and 7.6% of the C, while no positive values were observed in the N group. The percentage of hyperviscous samples was higher in the low sperm motility samples than in the normal group. Data on concentration of the biochemical markers seem to be decreased in asthenozoospermia. Pure and combined asthenozoo-spermia showed different behavior in sperm recovery after swim-up. Two different asthenozoospermias could be defined: the pure one where sperm environment is involved (immunological factor, hyperviscosity, and secretory gland function) and the combined, where the testis is comprised.     

Associations between biochemical components of human semen with seminal conditions

The aim of the study was to assess whether abnormal levels of seminal biochemical components could be associated with semen alterations and infertility. In this study, 92 human ejaculates from selected men were analyzed. Albumin, estradiol, ferritin, total proteins (TP), folic acid (FA), vitamin B12, alkaline phosphatase (ALP), creatine kinase (CK), gamma-glutamyl transpeptidase (GGT), lactate dehydrogenase were evaluated. Semen parameters and biochemical components of the 92 samples were correlated bySpearman’s rho coefficient. Albumin showed a negative correlation with sperm progressive motility and vitality (P < 0.05), CK with sperm concentration and vitality (P < 0.05), ferritin with sperm morphology (P < 0.05). FA negatively correlated with sperm concentration (P < 0.05) and GGT with sperm motility (P < 0.05). The values of biochemical components were compared for each semen parameters (concentration, motility, morphology, vitality) in samples ≤5th percentile with those >5th percentile and in patients with/without leukocytospermia, presence/absence of germ cells, increased/normal viscosity by Mann Whitney U test. The albumin (P < 0.001) and TP (P < 0.05) levels and the GGT activity (P < 0.001) were significantly higher in patients with sperm motility ≤5th percentile. Patients with sperm vitality ≤5th percentile showed increased albumin concentration (P < 0.01) and the CK activity (P < 0.001). The presence of germ cells in semen was concomitant with high values of ferritin (P < 0.01); the ALP activity (P < 0.01) and FA level (P < 0.001) were decreased in hyperviscous semen. The FA and estradiol levels were significantly decreased in the smoker group compared to those measured in the non-smoker group. Subjects were grouped in infertile patients and men with unknown reproductive potential. Infertile patients albumin and ferritin were significantly increased (P < 0.05). This study suggests that some biochemical components may be associated with human seminal pathological conditions.

Abbreviations: ALP: alkaline phosphatase; LDH: lactate dehydrogenase; GGT: γ-glutamyl transferase; CK: creatine kinase; ACP: acid phosphatase; ALB: albumin; TP: total proteins; FERR: ferritin, E: estradiol; FOL: folic acid; B12: vitamin B12; FSH: follicle stimulating hormone; LH: luteinizing hormone; T: testosterone; BMI: body mass index; WHO: World Health Organization.