The effect of varicocele repair on optimized sperm penetration assay.
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The aim of this study was to evaluate postoperative changes in sperm chromatin heterogeneity in varicocele patients. In 15 infertile patients with varicocele, sperm parameters including concentration, motility, and morphology were evaluated before and after surgical correction of varicocele. Sperm motion analysis using computer-assisted semen analyzer (CASA) was also performed. To analyze the sperm nuclear proteins, the acridine orange staining method was used. On semen analysis, sperm concentration and motility significantly increased after surgery (p = 0.002, p = 0.003, respectively), although sperm morphology was unaltered postoperatively. CASA parameters, including velocity, linearity, amplitude of lateral head displacement and beat cross frequency were unaltered postoperatively. On the other hand, acridine orange staining significantly increased postoperatively (p = 0.002). Varicocele influences the sperm chromatin condition, as well as sperm concentration and motility.
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ABSTRACT: Increased DNA fragmentation is found in sperm from infertile men. Varicocele is an important cause of male infertility, even though it is present in 15% of men who father children. Semen analysis does not always identify infertility in these patients. Sperm motility is strongly correlated with male fertility potential. The goal of this study was to determine the correlation between apoptosis and kinematics in the ejaculated spermatozoa of patients affected by varicocele. Fresh semen samples were obtained from 30 patients with varicocele and 15 fertile controls. These samples were compared using computer‐assisted semen analysis and were assayed to determine the degree of sperm apoptosis. The apoptotic index (AI) was calculated by dividing the number of terminal deoxynucleotidyl transferase—mediated deoxyuridine‐5′‐triphosphate nick end labeling (TUNEL) stained spermatozoa by the total number of Hoechst 33258‐stained sperm cells for 300 sperm. Five microscopic fields were analyzed to obtain 5 AIs for each individual. Results demonstrated no significant difference in semen quality and sperm motion characteristics; however, a significantly higher AI (23.05% ± 4.07%: mean difference ± SE, 95% CI, 15.06%–31.03%, P < .0001) was identified in the varicocele group than in the fertile controls. We concluded that sperm apoptosis does not seem to correlate with semen quality and sperm kinematics and that apoptosis is increased in ejaculated spermatozoa in patients with varicocele compared to normal fertile men.
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To examine and analyze semen quality and sperm ultrastructural characteristics of infertile patients with varicocele.This study included 118 infertile patients with varicocele (the VC group) and 76 normal semen donors (the control group). We obtained routine semen parameters, seminal plasma biochemical markers and the levels of reproductive hormones in the subjects, and observed the changes in sperm structure under the scanning electron microscope and transmission electron microscope.Compared with the normal control, the VC patients showed significantly decreased sperm concentration, sperm progressive motility, sperm viability (P < 0.05), but no remarkable difference in semen volume and non-progressive motility (P > 0.05). The concentrations of zinc and alpha-glycoside enzyme in the seminal plasma were markedly reduced in the VC group in comparison with the controls (P < 0.05), but there was no significant difference in the level of fructose (P > 0.05), nor in such seminal plasma biochemical markers as FSH, LH, T and E2 between the two groups (P > 0.05). The percentage of morphologically normal sperm was dramatically lower in the VC than in the control group ([56.76 +/- 15.32]% vs [12.34 +/- 6.58]%, P < 0.05), and the sperm deformities were mostly in the head and neck, mainly tapering pin head accompanied by complex abnormal differentiation.This study demonstrated that VC may lead to oligo-astheno-terato zoospermia, and hence male infertility, which may be attributed to the changes of seminal plasma microenvironment and sperm ultrastructure.
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To detect sperm mitochondrial membrane potential (MMP) of varicocele patients and investigate its clinical significance.Sixty-seven varicocele patients were divided into a VC1 (grade 1, n = 26), a VC2 (grade 2, n = 21) and a VC3 group (grade 3, n = 20). And 29 normal fertile volunteers were included in a control group ( m = 29). Conventional semen analyses were performed by computer-assisted semen analysis (CASA). Semen samples were washed, followed by JC-1 staining to evaluate the sperm MMP (JC-1+ %) by flow cytometry.The sperm MMPs of the VC1, VC2 and VC3 groups were siginificantly lower ([56.29 +/- 16.32]%, P < 0.05; [45.04 +/- 13.21]%, P < 0.01; [31.63 +/- 12.91]%, P < 0.01) than that of the control ([76.21 +/- 13. 96]%). There was a significant positive correlation between the percentage of JC-1+ and that of grade (a + b) sperm (r =0.693, P=0.000).The decreased MMP in the sperm of varicocele men might be one of the important causes of male infertility.
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Objective To explore the relationship between DNA damage and the expression of seminal plasma protein(SPP)in infertile men with varicocele. Methods Applying sperm chromatin dispersion(SCD)test,semen samples of 42 infertile men with varicocele-22 with abnormal semen analysis(group B)and 20 with normal semen analysis(group C)-and 20 men of normal fertility(group A)were detected for their sperm DNA integrity,and the expressions of SPP in each group were detected by two-dimensional gel electrophoresis(2-DE)and mass spectrometry(MS). Results Compared with group A,the sperm density and vitality in groups B and C decreased,and the sperm DNA integrity increased,the differences being significant(F=31.38-785.17,q=4.21-55.99,P0.05),a comparison between groups C and B showed that sperm density and vitality in group B declined,and sperm DNA integrity increased,the differences being significant(q=6.77-28.87,P0.05).Compared with group A,the sperm activity rate in groups B and C declined(H=45.79,P0.05),a comparison between groups C and B showed a decrease of sperm activity rate in group B(Z=-5.47,-4.03;P0.01);compared with group C,the sperm viability of group B was signi-ficantly lower(Z=-5.14,P0.01).Fifteen protein spots of interest were submitted to mass spectrometry and eight sports su-ccessfully identified,these proteins were associated with motility of sperm and capacitation,immune function,DNA damage,apoptosis and molecular transportation,of which,DNA-damage-associated proteins-SMG-1and IGFBP-3-expressed only in group B. Conclusion Sperm DNA damage due to varicocele might be an important cause of male infertility.
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Smoking and varicocele are frequent findings in the medical history and physical examination of patients attending and rological outpatient departments. However, data about their influence on human semen parameters, such as sperm concentration and motility, are contradictory. Therefore, the purpose of this study was to examine sperm function (acrosin activity and induction of the acrosome reaction) in smokers (n = 130) and varicocele patients (n = 30)compared with normal fertile donors (n = 20). The acrosomere action was detected by triple staining after 3 h ofincubation at 37°C, followed by treatment with 0.1%dimethyl sulphoxide (spontaneous acrosome reaction) and 10 μM calcium ionophore A23187 (induced acrosome reaction) for 1 h at 37°C. Acrosin activity was measured by gelatinolysis. The diameters around the sperm heads after gelatinolysis and the percentages of spermatozoa showinghalo formations were evaluated. The inducibility of the acrosome reaction was significantly lower in semen samples from smokers than in those from the fertile group (7.1 ±3.2 versus 11.2 ± 4.0%, P < 0.01), whereas no statistically significant difference was demonstrated in spermatozoa from patients with varicocele (9.3 ± 4.3%). Both the percentages of spermatozoa with halo formation (53.3 ±20.0 versus 76.6 ± 13.6%, P < 0.05) and the halo diameters (16.1 ± 6.6 versus 31.0 ± 14.5 urn, P < 0.001) were significantly lower in the varicocele group than in thesamples from fertile men. These data suggest that smoking and varicocele affect sperm function, and that the standard semen parameters alone are insufficient to evaluate the influence of both factors on human male fertility.
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Objective:To explore the levels of sperm DNA damage in infertile males with varicocele and the effect of high ligation of the spermatic vein(HLSV) on DNA damage in varicocele(VC) patients,and investigate the test of sperm DNA damage to evaluate the feasibility of the VC patients' fertility and the effect of HLSV. Methods:Semen samples from 58 patients with clinical varicocele and 30 normozoospermic donors were examined.Varicocele sperm samples were classified as normal or abnormal according to World Health Organization guidelines.Sperm DNA damage was evaluated by the sperm chromatin dispersion(SCD) text.24 VC patients underwent HLSV,each of them underwent sperm analysis and DNA damage evaluation before and 3 months after HLSV. Results:DNA damage levels were significantly greater in patients with varicocele,either with normal(23.94±6.04) or with abnormal(33.64±7.11) semen profile,compared with controls(10.78±5.01)(P0.001).Compared with pre-operation,HLSV achieved a significant decreases in the percentage of sperm DNA damage(P0.001). Conclusions:The presence of a varicocele is associated with high levels of DNA damage spermatozoa.HLSV can effectively decreases in the percentage of sperm DNA damage.For VC patients' fertility and the effect of HLSV,the test of sperm DNA damage is a good and feasibility evaluation mark.
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Spermatic Vein
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Background: Aim of current study was to determine whether the Sperm Penetration Assay (SPA) can be used as a test to discriminate the infertile male from fertile one. We have also correlated the SPA with semen analysis.Methods: Sperm characteristics namely Semen analysis and the sperm penetration assay were tested in 44 infertile and 10 fertile men. Sperm penetration assay was determined by using zona free hamster eggs.Results: With decreasing spermatozoa concentration in the semen there was significant decrease in percentage penetration of zona free Hamster eggs (p<0.001). There was decrease in Sperm penetration assay with deteriorating progressive sperm motility (p<0.05) and no consistent relationship appeared between the sperm morphology and the Sperm penetration assay (p>0.05). Conclusions: The Sperm penetration assay could discriminate the infertile group from fertile group significantly (p<0.001). The test appeared to be highly reproducible and probably identifies a truly infertile male.
Penetration (warfare)
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Penetration rate
Sperm washing
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