Sperm density measurement: should this be abandoned?

Sperm density measurement by means of the Makler chamber was performed on a single semen specimen from 2 groups: 104 proven fertile men and 53 infertile men; 11.5% of the fertile population had a sperm density less than 20 million/ml (median 84, mean 91.3, SD 60.5) and 33% of the infertile population had a sperm density greater than 20 million/ml (median 10, mean 26.9, SD 34.8). There was a statistically significant difference in median between the 2 groups, but when a discriminant analysis was applied, sperm density could predict fertile status with only 68% accuracy at optimum cut-off. Sperm density is valueless in distinguishing fertile from infertile men and is neither useful in diagnosis nor in monitoring the progress of treatment for male infertility. It should be abandoned as a quotable measurement of male fertility.     

Sperm morphology: unstained and stained smears in fertile and infertile men

Two semen specimens from 34 fertile donors and 82 male partners of infertile couples were evaluated for semen quality. Sperm morphology was evaluated with unstained specimens and following Papanicolaou staining. A significant difference (p less than 0.001) was found in the percentage of morphologically normal spermatozoa in stained and unstained semen smears. Fertile semen donors had a significantly (p less than 0.001) higher percentage of normal sperm than men of unproven fertility. When compared to the fertile donors, 78% of the infertile men had increased teratospermia. Papanicolaou staining appears to enhance sperm morphology and allow improved discrimination of sperm head abnormalities.     

Diagnostic value of sperm function tests and routine semen analyses in fertile and infertile men

The results of routine semen analyses, the zona-free hamster oocyte penetration test, the hypoosmotic swelling test, and semen adenosine triphosphate levels were studied in 66 fertile and 130 infertile men. Multivariate discriminant analysis demonstrated that routine semen parameters including semen volume, sperm count, percent sperm motility, and percent normal spermatozoa in combination could predict the fertility of these patients with 70.4% accuracy. Of the three sperm function tests evaluated, the zona-free hamster oocyte penetration test and the hypoosmotic swelling test were selected by the multivariate discriminant analysis as variables capable of providing significant information on the fertility status of the patients. However, the addition of the results of these two tests to the routine semen analysis did not significantly improve the predictability of fertility. The overall correct prediction rate was 77.6% after incorporation of the results of these two sperm function tests. In this group of subjects, the presently available sperm function tests did not predict the fertility status of a patient with a high degree of accuracy.     

精索静脉曲张不育患者的精液质量和精子形态学观察

目的:观察不育伴精索静脉曲张(VC)患者的精液质量和精子形态学变化。方法:98例不育伴VC患者精液按WHO标准常规分析并对精子形态学进行评价。130例正常供精者精液检测结果作为对照。结果:VC患者正常形态精子和前向运动精子明显低于对照组(P<0.001),精子畸形的类型以梨形、锥形和不定型头部畸形为主。结论:VC可导致精子畸形率升高,后者可能是男性生育力受损的重要标志之一,经染色后的精子形态学分析是判定VC患者精子受损的一个敏感指标。     

Time-related decline in sperm motility patterns in men with cytotoxic antibodies

Sperm motility patterns in semen from 10 fertile nonautoimmune men (fertile control group) and 33 infertile men with various titers of cytotoxic sperm antibodies in their seminal plasma (group 1: titers less than or equal to 16, n = 6; group 2: titers 64 to 512, n = 12; group 3: titers greater than or equal to 1024, n = 15) were evaluated every 2 hours for 12 hours and finally at 24 hours. A significant decline in sperm swimming speed and linearity was observed at 6 hours in semen from 27 infertile men with sperm antibodies. Beginning at 8 hours, semen from sperm antibody-positive men in group 2 showed a significant decline in percentage motile sperm (p less than 0.001) compared to the fertile controls. The percentage motility in semen of donors in groups 1 and 3 was significantly lower than that in semen of fertile donors at 10 hours (p less than 0.05), 12 hours (p less than 0.01), and 24 hours (p less than 0.001). The mean velocity in groups 2 and 3 was significantly less than that in fertile controls at 10 and 12 hours (p less than 0.05). The linearity of sperm motility started to decline 4 hours after semen samples were obtained from sperm antibody-positive groups 2 and 3 in contrast to sperm antibody-negative fertile or infertile groups (p less than 0.05). It is concluded that the presence of cytotoxic sperm antibodies in the seminal plasma adversely affects sperm linearity as early as 4 hours after semen collection.(ABSTRACT TRUNCATED AT 250 WORDS)     

Phospholipid composition of human sperm and seminal plasma in relation to sperm fertility

The phospholipid and fatty acid composition of sperm was studied in 8 healthy and 16 infertile men. Infertile men randomly formed from the patients with normal semen parameters according to WHO criterion. Therefore, all semen parameters of infertile patients were similar to the same characteristics of the semen of healthy men, except the abnormal forms. The amount of abnormal forms in infertile men was significantly higher than in healthy men. Sperm from infertile men show a drastic loss of phosphatidyl ethanolamine. At the same time, the significant increase of phosphatidyl serine in the sperm and seminal plasma of sterile patients was found. Lysophosphatidyl serine in the sperm of the infertile men was detected. Fatty acid composition of the semen of infertile men was altered. The levels of stearic and n-3 polyunsaturated fatty acids (eicosopentaenoic and docosahexaenoic acids) was dramatically lowered, but the values of some n-6 polyunsaturated fatty acids (linolenic and docosatetraenoic) acids increased. There was significant positive correlation between docosahexaenoic acid and sperm motility (r = .82, p < .001) and negative correlation between linolenic acid and spermatozoa motility (r = -0.58. p < .05). Infertility of men with normal semen quality can originate from the disorder of sperm lipid metabolism. The drastic loss of phosphatidyl ethanolamine and n-3 polyunsaturated fatty acids with simultaneous enhancement of phosphatidyl serine and some n-6 polyunsaturated fatty acids in sperm could be an important cause of male infertility.     

Male factors and the likelihood of pregnancy in infertile couples. I. Study of sperm characteristics

A prospective study of 394 infertile men was conducted over 3 years following a primary semen analysis. The cumulative pregnancy rate was 43 and 64% after 1 and 3 years, respectively. The pregnancy rate was significantly higher in the secondary infertile group. The study of various sperm factors and the occurrence of pregnancy showed that they were not of equal significance in predicting male fertility potential. The percentage of pregnancies decreased significantly only when the sperm concentration was less than 5 x 10(6)/ml. The pregnancy rate increased significantly with the percentage of motile sperm. The percentage of sperm with normal morphology was also found to be significantly higher when a pregnancy occurred than when the couple remained infertile (43.6% vs 37.7%). In a detailed morphological analysis of the sperm, six abnormalities (microcephaly, double head, amorphous head, cytoplasmic droplet, bent tail and coiled tail) were found to be significantly more frequent when a pregnancy did not occur. The most predictive value was given by the Multiple Anomalies Index (MAI), which is the mean number of abnormalities observed per abnormal sperm. The pregnancy rate was significantly lower after both 1 and 3 years when the MAI was greater than 1.6. Multivariate analysis showed that the best prognostic indicator of fertility was given by the percentage of motile sperm and the MAI, particularly in patients with primary infertility.     

A critical method of evaluating tests for male infertility

Considerable uncertainty surrounds the selection of test values that separate infertile from fertile men in the evaluation of male infertility. We herein describe an objective method of determining these values, referred to as threshold values, for different infertility tests. Using test results from fertile men threshold values were chosen such that 96 per cent of the semen samples from the fertile men were scored as fertile. These threshold values then were used to evaluate 100 semen samples from 74 men presenting for evaluation of infertility. Using this method we constructed infertility profiles on each of the 100 semen samples presented for infertility evaluation and found that the zona pellucida-free hamster egg penetration test (a measure of a spermatozoon's ability to undergo capacitation and penetrate an egg) identified 66 per cent of these samples as infertile, while multiple exposure photomicrography (a quantitative measure of sperm motility) identified 54 per cent of these samples as infertile. This compares with results from routine semen analyses using the same method, which identified none of the samples as infertile by sperm motility grade, 1 per cent by semen pH, 4 per cent by the percentage of motile sperm, 7 per cent by the total count of motile sperm, 10 per cent by the total sperm count, 11 per cent by the semen leukocyte concentration, 12 per cent by the concentration of motile sperm, 13 per cent by ejaculate volume, 16 per cent by sperm concentration and 27 per cent by sperm morphology. This method of analyzing infertility test results provides insight into the potential causes of male infertility and offers a critical approach towards understanding the complex problem of male fertility dysfunction.     

Incidence of chromosome 8,10,X and Y aneuploidies in sperm nucleus of infertile men detected by FISH

We studied the frequency of aneuploidy in sperm nuclei of six infertile men with abnormal semen profile and normal karyotype, using fluorescence in situ hybridization (FISH) with DNA probes for chromosomes 8, 10, X and Y. The control group consisted of four healthy fertile men with normal karyotype and semen profiles. The purpose of this study was to determine whether there are differences between infertile male donors and control donors for: (1) the incidence of sex chromosome aneuploidy, and (2) the number of disomies for chromosomes 8, and 10 cosegregating with chromosomes X and Y. FISH analysis showed no significant differences of sex ratios of the sperm nuclei in and between infertile and control groups. The most significant abnormalities in the infertile group were clusters of sperm nuclei bearing XY and XYY. In addition, the incidence of disomic sperm nuclei for chromosomes 8 and 10 consegregating with sex chromosomes was not significantly different between the patient and control groups, nor within them. However, the total frequency of aneuploid sperm nuclei was significantly different between the infertile group and the control group. We observed a significant excess of sperm nuclei bearing chromosome 10 along with disomy for chromosome Y (10YY). In conclusion, our results from FISH analysis demonstrate a significantly increased frequency of aneuploidy for the sex chromosomes in sperm nuclei from infertile men. Therefore it may be concluded that infertility is a risk factor for sex chromosome aneuploidy in sperm nuclei.     

精子凋亡与男性不育关系初探

目的 :探讨精子凋亡与男性不育的关系。　方法 :应用流式细胞术 (FCM)检测正常生育与不育男性精液中精子凋亡情况。　结果 :在不同人群中精子凋亡均有一定的发生率 ,生育组凋亡率 (PAS)为 (4 2 8± 1 6 6 ) % ,不育组为 (18 6 7± 8 5 5 ) % ,两者差异极显著 (P <0 .0 1) ,且PAS与精液量、精子密度、精子前向运动率、精子正常形态率呈显著负相关 (P <0 .0 1)。　结论 :精子凋亡与男性不育有着十分密切的关系 ,FCM检测精子凋亡是一种迅速、准确、客观、可靠的评估精子功能和男性生育力的新方法     

Relationship between semen quality and seminal plasma components: alpha-glucosidase, fructose and citrate in infertile men compared with a normospermic population of Tunisian men

The aim of this study was to evaluate the correlation between the secretory function of the male accessory glands and sperm parameters in normospermic controls and infertile patients. One hundred and fifty-nine men were investigated: they were composed of two groups: normospermic ( n  = 37) and infertile ( n  = 122) men with altered sperm characteristics. These infertile men were divided into the following groups: asthenozoospermia ( n  = 38), teratozoospermia ( n  = 40) and asthenoteratozoospermia ( n  = 44). The patients underwent semen analysis and measurements of fructose, neutral alpha-glucosidase and citric acid. The level of fructose was significantly decreased in asthenozoospermic and increased in asthenoteratozoospermic men. It was significantly correlated with semen volume, sperm count, motility and morphology. The seminal alpha-glucosidase levels were significantly correlated with semen volume and pH and citric acid was significantly correlated with pH. Thus, alpha-glucosidase and citric acid levels were associated with semen pH. The significant correlation between semen parameters, accessory glands and epididymal functions highlights the relationship between semen and normal genital tract function.     

Sperm chromatin stability and zinc binding properties in semen from men in barren unions

Sperm chromatin stability and zinc binding properties were studied in semen samples from 115 men living in barren unions. Of these men, 26% had a high proportion of swelling sperm, i.e. less than 80% sperm with stable chromatin after exposure to the detergent sodium dodecyl sulphate. From 2-67% of seminal zinc was bound to high molecular weight ligands of vesicular origin (HMW). This shows that, among infertile men, liquefied seminal plasma has huge variations in zinc chelating properties. The relationship between prostatic palpatory status, the proportion of abnormal sperm, the percentage zinc bound to HMW (HMW-Zn), the time between ejaculation and analysis and chromatin stability were studied. Samples with low chromatin stability were found more frequently in men with low HMW-Zn levels in semen. The proportion of stable sperm decreased in samples with prolonged exposure to seminal plasma. Neither the proportion of stable sperm heads nor the percentage zinc bound to HMW could be used to predict the future chances of the infertile men fathering children when studied 15-180 min after ejaculation. To differentiate between initial zinc-dependent stability and superstability developed in seminal plasma, other more sensitive methods must be developed.     

Sperm DNA damage in men from infertile couples

Aim： To investigate the prevalence of high levels of sperm DNA damage among men from infertile couples with both normal and abnormal standard semen parameters. Methods： A total of 350 men from infertile couples were assessed. Standard semen analysis and sperm chromatin structure assay （SCSA） were carried out. Results： Ninety-seven men （28% of the whole study group） had a DNA fragmentation index （DFI） 〉 20%, and 43 men （12%） had a DFI 〉 30%. In the group of men with abnormal semen parameters （n = 224）, 35% had a DFI 〉 20%, and 16% had a DFI 〉 30%, whereas these numbers were 15% and 5%, respectively, in the group of men with normal semen parameters （n = 126）. Men with low sperm motility and abnormal morphology had significantly higher odds ratios （ORs） for having a DFI 〉 20% （4.0 for motility and 1.9 for morphology） and DFI 〉 30% （6.2 for motility and 2.8 for morphology） compared with men with normal sperm motility and morphology. Conclusion： In almost one-third of unselected men from infertile couples, the DFI exceeded the level of 20% above which, according to previous studies, the in vivo fertility is reduced. A significant proportion of men with otherwise normal semen parameters also had high sperm DNA damage levels. Thus, the SCSA test could add to explaining causes of infertility in cases where semen analysis has not shown any deviation from the norm. We also recommend running the SCSA test to choose the appropriate assisted reproductive technique （ART）.     

人类精子低渗试验指数分析

目的：探讨精子低渗试验指数（ＳＨＯＴＩ）对评估男性生育力的临床价值。方法：采用两种不同渗量的低渗液结合活体染色技术，对１２９例男性精液进行精子低渗试验。结果：生育组与不育组比较精子卷尾率、ｇ型肿胀精子率和精子低渗试验指数差异均显著（Ｐ＜０．０１），生育组ＳＨＯＴＩ分布范围为：０．０９６～０．３３６；不育组ＳＨＯＴＩ分布范围为０．０２５～０．２１５。结论：ＳＨＯＴＩ综合反映了精子的整体膜功能及受精能力，可作为评估男性生育力的一个新的指标。初步确定临床参考值为ＳＨＯＴＩ＜０．０９６，提示精子功能不良，不能生育；在０．０９６～０．２１５之间精子功能部分正常，有机会生育；在０．２１５～０．３４０之间预示精子功能正常，生育力良好。     

Effect of abamectin exposure on semen parameters indicative of reduced sperm maturity: a study on farmworkers in Antalya (Turkey)

Environmental exposure to pesticides may cause serious health risks including fertility and reproductive function. The aim of this study was to highlight whether there is a relationship between exposure to abamectin and male fertility parameters of farmworkers. Twenty male farmworkers who were using abamectin and 20 men not exposed to pesticides were recruited as experimental and control groups, respectively. Semen analysis, molecular markers of sperm maturity and serum reproductive hormone levels were evaluated. In experimental group, high plasma abamectin levels were detected. These men have decreased sperm motility. Moreover, diminished molecular markers of sperm maturity, such as decreased hyaluronic acid (HA) binding of sperm, increased numbers of aniline blue positive sperm and increased percentage of creatine kinase (CK) positive sperm, were observed in abamectin‐exposed men. Their serum testosterone, LH and FSH levels did not change significantly. We conclude that exposure to abamectin may impair male fertility by effecting semen quality.     

Evaluation of HSPA2 in fertile and infertile individuals

Heat‐shock protein A2 (HSPA2) is a testis‐specific member of the HSP70 family known to correlate with sperm maturity, function and fertility. The aim of this study was to compare expression of HSPA2 in fertile and infertile individuals using a recently marketed highly purified polyclonal antibody. Thus, after analysing sperm concentration, motility and morphology in semen sample of 49 individuals with male factor infertility and 47 fertile individuals according to World Health Organization guidelines, we evaluated HSPA2 by microscope florescence, flow cytometry, Western blot and RT‐PCR. We observed higher percentage of sperm expressing HSPA2 in anterior and equatorial regions in fertile than infertile individuals by microscope fluorescence. Percentage of sperm expressing HSPA2 in two conditions (fixed permeabilised and capacitated) by flow cytometry showed that percentage of HSPA2‐positive sperm in fixed permeabilised and also capacitated samples was significantly different between fertile and infertile individuals. Western blot analysis of HSPA2 in semen samples revealed high variation within the fertile and infertile population. The results of RT‐PCR revealed higher expression of HSPA2 in the fertile compared to infertile individuals, but this difference was not significant. According to the results, we suggest that HSPA2 expression is heterogeneously expressed on different part of fixed permeabilised sperm and its expression is significantly higher in fertile compared to infertile individuals. The surface expression of this protein significantly increases following capacitation in both fertile and infertile individuals. HSPA2 expression significantly correlates with sperm concentration and morphology. Therefore, aberrant HSPA2 expression may play an important role in capacitation and fertilisation processes.     

精浆游离L-肉毒碱水平及其与精子密度、活动率及活力的相关性研究

目的:研究正常生育及不育男性精浆中游离L-肉毒碱水平差异及其与精子密度、活动率(a+b+c级精子百分率)及活力(a+b级精子百分率)之间的相关性,探讨精浆中游离L-肉毒碱水平对男性生育力的影响及其在不育症检查和治疗中的作用。方法:分别采用高效液相色谱法和计算机辅助精液分析系统,测定了230例不育症患者(精子密度正常117例,少精子症81例,无精子症32例)和30例正常生育男性精浆中游离L-肉毒碱水平及精子密度、活动率、活力等参数。根据检查结果对不育症患者分组后,以SPSS12.0软件包进行统计学分析,比较各组间游离L-肉毒碱水平的差异以及游离L-肉毒碱水平与精子密度、活动率、活力之间的相关性。结果:正常生育组精浆游离L-肉毒碱水平明显高于不育组(P<0.01)。精液中精子密度越低、活力越弱,这种差异性越显著。相关性分析结果显示,精浆游离L-肉毒碱水平与精子密度呈显著正相关关系(r=0.521,P<0.01),与精子活动率和活力之间也具有正相关关系(r=0.319,P<0.01;r=0.251,P<0.01)。结论:精浆L-肉毒碱水平与精子密度、活动率和活力之间密切相关,其含量测定作为一项有用的生化指标,可为男性不育症检查及临床诊治和进行有关男性生殖功能机制研究提供参考。     

Heterologous ovum penetration test and seminal parameters in fertile and infertile men

Sperm penetration rates in the heterologous ovum penetration test were correlated with results of routine semen analysis in 30 fertile and 50 infertile men. There was no difference in penetration rates when comparing infertile men with normal and abnormal seminal parameters, nor was any difference seen between fertile (15-83%) and infertile men (8-83%). Of the 22 infertile men with normal seminal characteristics, seven had partners with no discernible reproductive dysfunction. The penetration rates of these men (38-81%) did not differ from those of fertile men. Stepwise regression analysis of seminal characteristics, with penetration as the dependent variable, indicated that sperm count and morphology are the most important seminal parameters for fertility assessment. Discrimination analysis revealed that sperm numbers and morphologic variables provide significant information for discriminating between fertile men and infertile men with normal or abnormal seminal characteristics. Sperm penetration and motility were not indicated as important factors. The present data suggest that, in unselected male patients seeking reproductive evaluation, the sperm penetration assay did not yield any additional information on the cause of infertility.     

Preoperative parameters related to the improvement of semen characteristics after surgical repair of varicocele in subfertile men

To know the parameter related to improvement of each semen characteristic (sperm density and percentage of progressive motile sperm) after high ligation of the left internal spermatic vein, 119 men from infertile couples were investigated. In each individual, the semen characteristic was considered to be different from the postoperative mean value if it was over or below 2 SD of the preoperative mean. Although no relationship was detected with the age of the patient, the duration of infertility or the preoperative semen characteristics, a definite relationship was found between the degree of varicocele and the improvement in each semen characteristic. For improvement of the sperm density, each of the volume and the score count by Johnsen's scale of the affected and unaffected testes and the serum hormone values (LH, FSH and testosterone) showed correlations. Meanwhile, for the improvement of the percentage of progressive motile sperm, correlations were found only with each of the testicular volume on the unaffected side and the serum testosterone level.     

生育与不育男性精子核DNA成熟度的比较研究

通过对１９对生育夫妇和７２对不育夫妇丈夫精液中绿色吖淀橙（ＡＯ）荧光精子（成熟精子）比率分布情况的比较分析，探讨了绿色ＡＯ荧光精子百分率与精液参数及精子授精能力之间的关系。在１～３年随访中，通过人工授精、其他治疗或在非治疗期，５４对绿色ＡＯ荧光精子≥５０％的夫妇有１８对怀孕，而１８对绿色ＡＯ荧光精子＜５０％的夫妇无１例怀孕。结果还显示了绿色ＡＯ荧光精子百分率与精液参数无关。认为该项指标从精子核ＤＮＡ成熟状态角度反映了单凭精液分析所不能反映的男性生育力。精液中绿色ＡＯ荧光精子＜５０％，是低生育力精液的评估界限