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Evaluation of a cut-off value for normal sperm morphology using strict criteria to predict fertilization after conventional in-vitro fertilization and embryo transfer in asthenozoospermia.

The predictive value of sperm morphology evaluation using strict criteria (MEUSC) on fertilization and pregnancy rates was evaluated in 60 couples undergoing in-vitro fertilization (IVF) and embryo transfer. In all, 47 of the male partners had a progressive sperm motility < 50% with a density > 20 x 10(6)/ml (asthenozoospermia) and 13 had normozoospermia. MEUSC was performed on the same semen sample that was used for IVF on at least 100 spermatozoa after Papanicolaou staining at x 1250 magnification. Defects in the head, mid-piece and tail were counted separately, borderline forms were classified as abnormal and the teratozoospermia index was calculated for each sample. Of the spermatozoa in the asthenozoospermic group, 13.0% were judged as normal on MEUSC versus 19.8% in normozoospermia (P = 0.0013). The number of normal spermatozoa on MEUSC correlated with the progressive motility in asthenozoospermia (rho = 0.41, P = 0.0043). Defects in the mid-piece (P = 0.0004) and tail (P = 0.025) were more common, and the teratozoospermia index (P = 0.015) was higher in asthenozoospermic than normozoospermic samples. The parameters of MEUSC did not correlate with the fertilization rate. In asthenozoospermia, differences in fertilization rates were calculated for each cut-off value between 4 and 10% normal spermatozoa on MEUSC. A cut-off value of 6% gave the best statistical power. If > 6% of spermatozoa were normal, a median of 66.7% of ova were fertilized, compared with a median fertilization rate of 35.4% if < or = 6% of spermatozoa were normal (P = 0.022). The highest cut-off value still giving significant discrimination was 8% (normal MEUSC > 8%, median fertilization rate 66.7%; < or = 8%, median fertilization rate 35.7%, P = 0.028), and this may be used as the critical value for normal morphology in asthenozoospermia. It is concluded that despite a significant linear relationship between morphology using strict criteria and progressive motility, MEUSC is still useful as a predictor for fertilization in asthenozoospermia. In cases of asthenozoospermia with < or = 8% normal spermatozoa on MEUSC, micromanipulation should be considered.

Adult↗

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.

Acid Phosphatase↗

Sperm mitochondria of patients with normal sperm motility and with asthenozoospermia: morphological and functional study.

Studies were performed on ejaculated human spermatozoa (32 subjects with normal sperm motility and 25 subjects with low sperm motility). Morphology of sperm midpiece was evaluated in light, fluorescent and transmission or scanning electron microscope. Changes in mitochondrial membrane potential (delta(psi)m) and mass of mitochondria were analysed by flow cytometry using mitochondrial specific probes JC-1 and Mito Tracker Green FM. Moreover, oxidoreductive capability of sperm mitochondria was assessed using cytochemical reaction for NADH-dependent dehydrogenases. In flow cytometry analysis of JC-1-stained spermatozoa, two asthenozoospermic subpopulations were distinguished: patients with a high percentage (76 +/- 11%, 13 subjects) and patients with a low percentage (29 +/- 14%,12 subjects) of spermatozoa with functional-polarized mitochondria with high delta(psi)m. Our microscopic investigations of spermatozoa of seven asthenozoospermic patients reveal that the deformed and unusually thickened sperm midpieces (50-70% of cells), occasionally with persistent cytoplasmic droplet, contain supernumerary mitochondria with normal substructure, full oxidoreductive capability and high delta(psi)m. The midpiece deformations cause nonprogressive movement or immotility. They can also appear in smaller number of spermatozoa (5-35% of cells) in patients with normal sperm motility. Moreover, in three cases of asthenozoospermia midpiece malformations were accompanied by abnormal morphology of outer dense fibers and axoneme. The cytochemical, fluorescence and SEM studies showed the absence of midpieces in many (60-80%) spermatozoa in some other cases of asthenozoospermia. The morphological observations corresponded with flow cytometry analysis of Mito Tracker Green FM-stained spermatozoa. Our results suggest that in some cases of asthenozoospermia the sperm mitochondria can be functionally active and display high delta(psi)m in large number of cells. The results may suggest that asthenozoospermia does not necessarily result from energetic disturbances of sperm mitochondria. The low sperm motility may be associated with deformations of the mitochondrial sheath containing functional mitochondria. The combination of fluorescence microscopy and flow cytometry with electron microscopic investigations is a sensitive, precise and comprehensive examination which helps discover sperm abnormalities responsible for asthenozoospermia.

Adult↗

Effect of sperm morphology and motile sperm count on outcome of intrauterine insemination in oligozoospermia and/or asthenozoospermia.

Eighty-six couples with long-standing infertility and poor postcoital test, due to oligozoospermia and/or asthenozoospermia (68 cases) or mucus hostility (18 cases), were treated by 411 intrauterine inseminations (IUI) with motile sperm suspensions from the husband's semen. The pregnancy rate per couple in the group with abnormal semen was lower than in the group with mucus hostility (22% versus 38.9%). Influence of seminal and other parameters on outcome of IUI was assessed by discriminant analysis, and a significant correlation with pregnancy rate was found for motile sperm count and sperm morphology. Teratozoospermia (normal morphology less than 50%) affected the outcome of IUI both when associated with moderate oligozoospermia and/or asthenozoospermia (motile sperm count greater than or equal to 5 X 10(6)/mL) (success rate per couple: 11.1%), and, even more, when associated with severe oligozoospermia and/or asthenozoospermia (motile sperm count less than 5 X 10(6)/mL), where no pregnancy was achieved. In the absence of teratozoospermia, the success rate per couple both in severe and in moderate oligozoospermia and/or asthenozoospermia had similar results (33.3% versus 35.7%). In conclusion, the absence of teratozoospermia appears to be an effective criterion for selecting couples with infertility due to oligozoospermia and/or asthenozoospermia who may benefit from IUI.

Adult↗

Identification of a Novel Splice-Site variant in TACR3 (c.888&#x2009;+&#x2009;1G&#x2009;>&#x2009;A) Associated with Asthenozoospermia and Hypogonadotropic Hypogonadism in an Iranian Family.

BACKGROUND: TACR3 encodes the receptor for neurokinin B, a key regulator of the hypothalamic-pituitary-gonadal axis. Disruption of this pathway can impair gonadotropin release and male reproductive function. Given the genetic heterogeneity of male infertility, this study aimed to identify novel variants in TACR3 that may underlie asthenozoospermia and related hormonal abnormalities. METHODS: Fifteen infertile men with confirmed asthenozoospermia were enrolled. Whole-exome sequencing (WES) was performed on genomic DNA from peripheral blood, and the candidate variant was validated by Sanger sequencing. Functional predictions were made using PolyPhen-2, SIFT, MutationTaster, and REVEL. TACR3 mRNA expression levels were assessed by real-time PCR in available samples. RESULTS: A novel splice-site variant, TACR3 (NM_001059.3:c.888&#x2009;+&#x2009;1G&#x2009;>&#x2009;A), was detected and found to segregate with infertility in one family, appearing homozygously in two infertile brothers and heterozygously in the proband with severe asthenozoospermia. The variant was absent in public and local genomic databases, suggesting its extremely rare frequency. Furthermore, RT-PCR showed a dramatic reduction or complete loss of TACR3 expression in affected individuals, confirming its deleterious effect on splicing and mRNA stability. CONCLUSION: We identified a previously unreported splice-site mutation in TACR3 (c.888&#x2009;+&#x2009;1G&#x2009;>&#x2009;A) that likely causes familial infertility by disrupting the neurokinin B/NK3R signaling pathway. While the heterozygous proband exhibited severe asthenozoospermia, the homozygous brothers displayed hormonal profiles typical of hypogonadotropic hypogonadism. These findings extend the mutational landscape of TACR3 and highlight its essential contribution to male reproductive endocrinology.

Humans↗

Subzonal sperm insemination and total or extreme asthenozoospermia: an effective technique for an uncommon cause of male infertility.

OBJECTIVE: To assess the value of subzonal insemination (SUZI) in cases of total or extreme asthenozoospermia. DESIGN: The results of IVF after SUZI were assessed in six cycles (four couples) with documented asthenozoospermia. SETTING: Institut de Médecine de la Reproduction, Marseille, France. PATIENTS: Four couples underwent six retrieval cycles. In all cases asthenozoospermia was total (100% of immotile spermatozoa) or extreme (5% nonprogressive motility). Oligozoospermia and teratozoospermia were ruled out in all cases. In two cases electron microscopy revealed the absence of the central singlet in the flagellum axonemal complex and in one case a major dysplasia of the fibrous sheath in the flagellum principal piece. RESULTS: The overall fertilization rate was 45%. The diploid embryo rate was 33%. Embryo quality was satisfactory. In five of six cycles, three embryos were transferred. Two triplet pregnancies were obtained corresponding to an implantation rate per transferred embryo of 40%. CONCLUSION: Total or extreme asthenozoospermia seems to be a good indication for SUZI.

Adult↗

Coenzyme Q(10) supplementation in infertile men with idiopathic asthenozoospermia: an open, uncontrolled pilot study.

OBJECTIVE: To clarify a potential therapeutic role of coenzyme Q(10) (CoQ(10)) in infertile men with idiopathic asthenozoospermia. DESIGN: Open, uncontrolled pilot study. PATIENT(S): Infertile men with idiopathic asthenozoospermia. INTERVENTION(S): CoQ(10) was administered orally; semen samples were collected at baseline and after 6 months of therapy. MAIN OUTCOME MEASURE (S): Semen kinetic parameters, including computer-assisted sperm data and CoQ(10) and phosphatidylcholine levels. RESULT(S): CoQ(10) levels increased significantly in seminal plasma and in sperm cells after treatment. Phosphatidylcholine levels also increased. A significant increase was also found in sperm cell motility as confirmed by computer-assisted analysis. A positive dependence (using the Cramer's index of association) was evident among the relative variations, baseline and after treatment, of seminal plasma or intracellular CoQ(10) content and computer-determined kinetic parameters. CONCLUSION(S): The exogenous administration of CoQ(10) may play a positive role in the treatment of asthenozoospermia. This is probably the result of its role in mitochondrial bioenergetics and its antioxidant properties.

Adult↗

Clinical characteristics and light and transmission electron microscopic sperm defects of infertile men with persistent unexplained asthenozoospermia.

OBJECTIVE: To determine, with the use of transmission electron microscopy (TEM), the proportion of patients with permanent unexplained asthenozoospermia (<30% motility) who have an abnormality of sperm axonemal and periaxonemal structures. DESIGN: Retrospective study. SETTING: A university-affiliated public hospital. PATIENT(S): Sixty-one infertile men whose semen was submitted to TEM analysis because of persistent unexplained asthenozoospermia. MAIN OUTCOME MEASURE(S): The results of quantitative TEM analysis of the tails of the spermatozoa. INTERVENTION(S): None. RESULT(S): Based on a comparison with the axonemal anomalies observed in nine fertile control patients, the infertile population was divided into three groups: group I, with no detectable axonemal defects (26.2%); group II, with axonemal anomalies in either the midpiece or the principal piece (29.5%); and group III, with axonemal anomalies in both the midpiece and the principal piece (44.3%). However, defects in the mitochondrial sheath, fibrous sheath, and sperm head (acrosomic and postacrosomic cap) were observed in at least 50%, 30%, and 50%, respectively, of the patients in each group. The proportion of dense fiber anomalies of the midpiece increased significantly from group I to group III. No differences were observed between the three groups in sperm characteristics, anamnesis information, or clinical data. CONCLUSION(S): In patients with persistent unexplained asthenozoospermia, the frequent association of periaxonemal anomalies with axonemal deficiencies strongly suggests that axonemal deficiencies are not the unique cause of decreased motility.

Adult↗

Application of Poisson distribution theory to the zona-free hamster oocyte penetration test to assess sperm function of men with asthenozoospermia.

Assessments of the penetrating potential of human spermatozoa were carried out using the zona-free hamster oocyte penetration test on 4 groups of subjects exhibiting normal fertility, idiopathic asthenozoospermia(less than 40% motility), asthenozoospermia associated with varicocele and oligoaesthenozoospermia (less than 20 X 10(6) spermatozoa/ml and less than 40% motility). When the Poisson model was used to correct the results of the in-vitro penetration experiments for differences in motile sperm concentration, significant differences were apparent between the normal fertile controls and all 3 categories of asthenozoospermic patient. Furthermore, the penetrating ability of the motile spermatozoa from patients presenting with a varicocele or oligoaesthenozoospermia was significantly less than that for the group in which asthenozoospermia was the only detectable defect. These results emphasize the practical significance of the Poisson model in the analysis of male fertility and demonstrate that the asthenozoospermic condition is associated with a significant reduction in the fertilizing potential of the motile spermatozoa.

Animals↗

[Therapeutic application of kallikrein in idiopathic oligo- and asthenozoospermia (author's transl)].

The addition of kallikrein to frozen semen after thawing increases the sperm motility in cases of moderate asthenozoospermia. In native semen maximal increase of motility is found two hours after the addition of kallikrein. In vivo an increase of sperm motility was noted in cases of oligozoospermia and asthenozoospermia during treatment with 600 KE kallikrein/day. The wives of 9 patients with asthenozoospermia became pregnant during kallikrein therapy.

Female↗

Correlation of asthenozoospermia with increased antisperm cell-mediated immunity in men from infertile couples.

To investigate whether cell-mediated immunity (CMI) against sperm and/or antisperm circulating antibodies are associated with poor semen quality, a leukocyte migration inhibition factor (LMIF) assay and an enzyme-linked immunosorbent assay (ELISA) were performed in groups of men from infertile couples, men from fertile couples and sperm donors. Twenty-five of 102 men (25%) revealed positive CMI against sperm and 10 (10%) had positive antisperm antibody titers in their sera. Fifteen of 28 asthenozoospermic men (53%) from infertile couples revealed positive antisperm CMI. The incidence of antisperm CMI was significantly increased (P < 0.05) in the infertile men with asthenozoospermia compared with the men from the other two groups (men from fertile couples and sperm donors). No significant differences between migration indices were seen when such a comparison was done for oligoasthenoterato- and teratozoospermics. The results indicate that increased antisperm CMI is associated with asthenozoospermia in a significant number of men from infertile couples. The importance of these findings is discussed.

Autoantibodies↗

Treatment of asthenozoospermia with zinc sulphate: andrological, immunological and obstetric outcome.

OBJECTIVES: The role of zinc therapy on infertility caused by defective sperm quality is examined. STUDY DESIGN: In this study, 100 men with asthenozoospermia were randomised into two groups--250 mg twice daily zinc therapy for 3 months and no therapy. The patients were followed up for another 6 months. Sperm parameters, circulating antisperm antibodies, sex hormones and T helper cytokines were evaluated before and after treatment for the two groups. RESULTS: There was significant improvement in the sperm quality; sperm count (P<.02), progressive motility (P<.05), fertilising capacity (P<.01) and a reduction in the incidence of antisperm antibodies (P<.01). Zinc/cadmium was higher in the zinc therapy group (P<.02). T helper cytokine, interleukin-4 level was significantly higher after zinc therapy (P<.02) while TNFalpha showed a significant decrease (P<.05). CONCLUSIONS: Zinc therapy has a role in improving sperm parameters in men with asthenozoospermia, probably through its membrane stabilising effect as an antioxidant and its effect on cellular and humoral immunity by reducing the levels of antisperm antibodies and TNFalpha and increasing that of IL-4.

Adult↗

Distinct expression pattern of cytokines in semen of men with genital infection and oligo-terato-asthenozoospermia.

PROBLEM: The objective of this study was to evaluate the possible relevance of cytokines in seminal plasma (SP) of patients with accessory gland infection and oligoterato-asthenozoospermia. METHOD OF STUDY: Semen samples were obtained by masturbation from 90 men and were examined for the presence of interleukin (IL)-2, IL-6, IL-8, IL-11 and soluble CD23 (sCD23) by enzyme-linked immunosorbent assay. Five groups were included: (1) fertile men (n = 20), (2) infertile men with varicocele and oligo-teratoasthenozoospermia (V-OTA, n = 20), (3) infertile men with genital infection and OTA (INF-OTA, n = 20), (4) infertile men with idiopathic testicular lesion and OTA (ITL-OTA, n = 20) and (5) infertile men with azoospermia (AZOO, n = 10). RESULTS: We found that the mean level of IL-2 was higher in SP from infertile men compared with SP from fertile men (P < 0.05). Mean levels of IL-6, IL-8, IL-11 in SP of INF-OTA were higher than that of all other groups (P < 0.05, P < 0.05, P < 0.001, respectively). However, no significant differences could be detected between other groups. A significant increase was noted in sCD23 levels in SP from men with ITL-OTA compared with all other groups (P < 0.01). We have not observed any correlations between IL-2, IL-6, IL-8, IL-11 and sCD23 levels in SP and semen parameters. Spearman's correlation coefficient revealed that there was a significant association between IL-6, IL-8, IL-11 levels in men with INF-OTA. CONCLUSION: The measurement of each cytokine separately in the SP of men with INF-OTA, in spite of the existing significant differences, does not have a diagnostic value in male infertility. However, a combined determination of IL-6, IL-8, IL-11 in the SP of men with genital infection and oligo-terato-asthenozoospermia may provide clinically useful information for the diagnosis of male accessory gland infection.

Adolescent↗

[Microinjection of spermatozoa into the oocyte. An experimental model for assisted conjugation in oligo-asthenozoospermia as a cause of sterility].

This study investigates the possibilities to treat oligo-asthenozoospermia, a cause of sterility, by assisted conjugation. After completing acrosome reaction, normal motile spermatozoa are microinjected into mouse oocytes. After 2 hours of incubation time in medium with lactate and albumins, spermatozoa are put into dibutyryl 3',5'-guanosine monophosphate (dbcGMP) and imidazole for the completion of an acrosome reaction. One motile spermatozoon each was injected microsurgically into the perivitelline space of an oocyte. The survival rate of the oocytes after injection was around 85%. The oocytes with two pronuclei and a second polar body were considered as fertilized. Of those injected oocytes 21.5% were fertilized with acrosome reacted spermatozoa, whereas only 5.6% were fertilized with spermatozoa without dbcGMP treatment. Thus, motile and acrosome reacted spermatozoa essentially fertilize oocytes at microinjection into the perivitelline space very well. This technique may become a hopeful procedure to treat human oligo-asthenozoospermia in an IVF/ET-programme.

Animals↗

Laboratory methods for the diagnosis of asthenozoospermia.

Human sperm samples from 46 men were evaluated for standard semen parameters by two trained technicians, according to WHO criteria. A computer-aided semen analyzer (CASA, HTM-IVOS) was employed. The overall mean coefficient of variation for the 2 participants and the 46 samples studied was 17% for the percentage of progressive motile spermatozoa (r = 0.77). Twenty-nine (63%) samples were classified as asthenozoospermia by both methods. From the studied samples, 12 (26%) were classified as asthenozoospermia by the subjective method and 2 (4.3%) by CASA (p =.01). The two methods do not provide directly comparable data.

Humans↗

Pentoxifylline (BL 191) by oral administration in the treatment of asthenozoospermia.

The aim of this trial was to study the effects on semen of pentoxifylline administered chronically per os to patients with normogonadotropic asthenozoospermia. Fifteen infertile men (age 22 to 44 years) were incorporated. Each subject was submitted to a treatment with pentoxifylline per os at a dose of 1,200 mg per day during no less than four months (x plus or minus SE 6.6 plus or minus 0.7 months). Assessment of the results was performed by serial spermatograms performed during and up to three months after treatment. The results showed a significant improvement of the percentages of forwardly progressive spermatozoa and of live and motile spermatozoa. Five of the patients achieved a normalization of semen (frank improvement) and seven a significant improvement in comparison to the initial values. Pregnancy was obtained by two patients. Pentoxifylline could be a useful alternative for the treatment of patients with normogonadotropic asthenozoospermia.

Administration, Oral↗

Further studies on the effects of pentoxifylline on sperm count and sperm motility in patients with idiopathic oligo-asthenozoospermia.

To further investigate the effectiveness of pentoxifylline (trental) treatment in male infertility, we studied 22 young men (mean age 28.4 years) with "idiopathic" oligo-asthenozoospermia treated for 6 months with the drug (1200 mg daily orally). Sperm concentration and sperm motility were determined before therapy, as well as after 3 and 6 months of pentoxifylline administration. Moreover, fructose concentrations in seminal fluid and sperm ATP levels were assayed before and at the end of the treatment in five semen samples. Pentoxifylline therapy significantly increased both sperm concentration and sperm motility. Sperm concentration showed a 1.5-fold increase (p less than 0.01) at the 3rd month of therapy, and a 2.0-fold increase (p less than 0.001) at the 6th month, whereas sperm motility increased by 1.8-fold (p less than 0.001) and by 2.8-fold (p less than 0.001) respectively. At the end of the treatment, fructose concentrations in seminal fluid were significantly higher (p less than 0.001) than pretreatment values; in contrast, sperm ATP levels showed a significant (p less than 0.05) fall. These results suggest that pentoxifylline, probably acting on the cAMP metabolism, may be an useful drug in the treatment of idiopathic oligo-asthenozoospermia.

Adenosine Triphosphate↗

Changes in semen quality and in vitro sperm capacitation during various frequencies of semen collection in dogs with both asthenozoospermia and teratozoospermia.

Eight male dogs with both asthenozoospermia and teratozoospermia were used in this study. In experiment 1, semen was collected 10 times at intervals of 48 hr, 24 hr and 12 hr in 4 of the 8 dogs, and the semen quality was evaluated. In experiment 2, semen was collected 5 times at 24-hr intervals in the other 4 dogs. The spermatozoa collected on day 1 and day 5 were incubated for 4-6 hr in Canine Capacitation Medium, and the percentages of hyperactivated sperm (%HA), acrosome-reacted sperm (%AR), and the zona pellucida-binding sperm count (ZP sperm count) were assessed. The results of experiment 1 showed that the percentage of motile sperm increased and the percentage of abnormal sperm decreased markedly as the intervals between semen collections became shorter. When semen was collected at 12-hr intervals, the percentage of motile sperm increased from about 65%, the value before frequent collection was started, to about 80%, and the percentage of sperm with abnormal tails decreased from 30% to 15%. In experiment 2, the percentages of HA and AR, and the ZP sperm count in specimens collected on day 5 were higher than those in specimens collected on day 1, and the differences in % HA and in ZP sperm count were significant (P<0.05). The results demonstrated that sperm motility, abnormality, and potential fertility in dogs with asthenozoospermia and teratozoospermia can be temporarily improved by frequent semen collection.

Acrosome↗