Systemic sperm autoimmunity in spinal-cord injured men

Historically, spinal-cord injured men have been considered virtually sterile because of ejaculatory dysfunction commonly resulting from their injury. Assisted ejaculatory techniques, however, have overcome the problem of sperm transport and have allowed both the establishment of pregnancy through artificial insemination and the assessment of their semen quality. Most studies have noted the presence of asthenozoospermia in the setting of normal sperm concentration following electroejaculation or vibratory stimulated ejaculation. Thus far, little attention has been given to the basis for the frequent finding of asthenozoospermia, and the possibility of sperm autoimmunity in this group has not been adequately studied. In nine spinal-cord injured men, reproductive evaluation was performed consisting of hormonal measurements, testicular biopsy, and indirect immunobead tests for sperm autoimmunity. A mean sperm concentration was 144 +/- 185 x 10(6)/ml. However, the mean motile concentration was 33 +/- 62 x 10(6)/ml. Indirect serum immunobead showed positive IgG or IgA titers in 3 of 8 patients. Because of the disproportionately high incidence of an immunologic factor in spinal-cord injured men compared to able-bodied infertile men, sperm autoimmunity should be considered among the important causes underlying seminal dysfunction following spinal cord injury.     

Serum inhibin B as a marker for spermatogenesis

Inhibin B generated by Sertoli cells provides negative feedback on FSH secretion. In men, inhibin B seems to be the physiologically important form of inhibin. Serum inhibin B was measured by two-site immunoenzymatic assay in 40 normal men (27 years of age) with sperm concentrations 100 +/- 9.2 x 10(6)/mL, 51 subfertile men (31 years of age) with sperm concentrations 6.8 +/- 0.8 x 10(6)/mL, 16 men with varicocele with sperm concentrations 54.3 +/- 0.8 x 10(6)/mL (31 years of age), men with hypogonadotrophic hypogonadism, men with Klinefelter syndrome, and men with obstructive and non-obstructive azoospermia. In men with normal sperm concentrations (>20 x 10(6) mL) serum inhibin B was 201 +/- 17 pg/mL and FSH 4 +/- 0.5 IU/L. Varicocele patients showed normal sperm concentrations > 20 x 10(6)/mL, normal serum inhibin B (173 +/- 21 pg/mL), and normal FSH levels (4.6 +/- 0.6 IU/L). In patients with sperm concentrations < 20 x 10(6)/mL the inhibin B level was 118 +/- 14 pg/mL and the FSH level was 10 +/- 1.1 IU/L. In all patients, except those with hypogonadotrophic hypogonadism and Klinefelter syndrome. inhibin B and FSH were inversely correlated (r = -.41, p > 0.01). There was a positive correlation between inhibin B and sperm concentrations (r = .34, p < .01). In varicocele men there was a correlation of r = .574, p < .05. Inhibin B may be a marker of exocrine testicular function and may offer an improved diagnosis of testicular dysfunction.     

Evaluation of Hamilton-Thorn automated semen analysis system.

Sperm concentration and percentage motility values generated by the HT M-2030 system (CASA) were compared with those obtained by subjective semen analysis (SSA). Three calibrations arbitrarily designated as A, M, and D for gates and discriminators and three calibrations arbitrarily designated LI, MI, and HI for default pixel count (DPC) and default intensity (DI) were evaluated. The best correlation between CASA and SSA was observed utilizing M calibration (n = 114) with which sperm concentration was +0.3 x 10(6)/mL (r = .96) and motility was -6.3% (r = .89) compared to the values obtained by SSA. It was found that 35.9% of sperm concentration values and 34.2% of sperm motility values were within 10% of the values obtained by SSA. When sperm concentration was between 50 and 100 x 10(6)/mL the difference in motility was reduced (-3.2%) while the difference in sperm concentration was reversed (-2.6 x 10(6)/mL). LI calibration (DPC = 4, DI = 86) gave acceptable results with M calibration for sperm concentration (+2.1 x 10(6)/mL) and motility (-6.9%) compared to the values obtained by SSA. In the presence of sperm clumping, the difference between CASA and SSA was reversed for sperm concentration (+0.56 x 10(6)/mL for normal samples vs. -2.2 x 10(6)/mL for samples with clumping) and was reduced for sperm motility (-7.14% vs. -4.55%, respectively). HT M-2030 under proper calibration can be used as a rapid, objective, and reliable alternative to conventional semen analysis in routine and for research purposes.     

Have sperm counts deteriorated over the past 20 years in healthy, young Japanese men? Results from the Sapporo area

Changes in semen quality of healthy men is a controversial issue throughout the world. It is suspected that many chemical endocrine disrupters may affect the quality of semen. Although exposure to them may be extensive in Japan, no evidence of changes in semen quality has been reported. In this study, changes in semen volume and sperm counts were analyzed over 20 years in the Sapporo area of Japan. Semen volume and sperm counts were measured in 254 and 457 normal, healthy volunteers who lived in the Sapporo area in 1975-1980 and 1998, respectively. Posters and handbills were used to recruit participants in both studies. Semen samples were collected by masturbation after 3 days or more of abstinence. There was no change in semen volume between 1975-1980 and 1998. Mean sperm counts were 70.9 +/- 47.3 x 10(6)/mL in 1975-1980 and 79.6 +/- 49.3 x 10(6)/mL in 1998. Sperm counts did not decline over about 20 years. No significant correlation between age and sperm counts was recognized in either study. The rates of subjects with oligozoospermia and azoospermia were the same in both studies. In the 1975-1980 study, 34 of 254 (13.4%) participants had a child, and in the 1998 study, 51 of 457 (11.2%) participants had a child. Mean sperm count was significantly (P < .02) lower in the earlier study (66.0 +/- 44.9 x 106/mL) than in the 1998 study (98.7 +/- 60.2 x 10(6)/mL). This is the first reliable report in which changes in sperm counts in Japan were studied. We conclude that there was no evidence of deterioration in sperm counts of normal healthy men who lived in the Sapporo area of Japan over 20 years. However, selection bias in the recruitment of volunteers and the issue of variable abstinence might have affected the results of these studies. Therefore, well-designed prospective studies should be performed in several different regions to extrapolate our results on sperm counts to healthy, young Japanese men in general. Key words: Fertility, endocrine disruptors, seminalysis.     

Effect of body weight on testosterone/estradiol ratio in oligozoospermic patients

To evaluate the effect of body mass on the hormonal and semen profiles of subfertile men with oligozoospemia, sperm concentration and reproductive hormone levels were compared in two body mass index (BMI) groups: underweight or normal weight patients (BMI = 25 kg/m2) vs. overweight or obese patients (BMI > 25 kg/m2). The mean BMI was 27 +/- 4.6 kg/m2. The testosterone/estradiol ratio was significantly reduced in the high BMI group as compared to the low BMI group (17 +/- 4 vs. 12 +/- 3; p < 0.05). A similar difference was found in the sperm concentration (11.2 +/- 3.16 x 10(6)/ml vs. 8.1 +/- 2.6 x 10(6)/ml). A nonsignificant difference was found in the LH/FSH ratio (1.41 +/- 0.64 vs. 1.63 +/- 0.72). We concluded that obesity and the consequent estrogen excess decrease the sperm concentration by influencing the hypothalamo-pituitary system.     

Semen parameters in men with spinal cord injury： changes and aetiology

Aim： To assess the changes in semen parameters in men with spinal cord injury （SCI） and the possible causes of these changes. Methods： The study included 45 subjects with SCI. Semen retrieval was done by masturbation （2）, vigorous prostatic massage （n = 13）, penile vibratory stimulation （n = 13） or electroejaculation （n = 17）. Results： The semen of men with SCI showed normal volume （2.3 ± 1.9 mL） and sperm count （85.0 × 10^6 ± 83.8 × 10^6/mE） with decreased motility （11.6% ± 10.1%）, vitality （18.5 % ± 15.2%） and normal forms （17.5 ± 13.4%）, and pus cells has been increased （6.0 × 10^6 ± 8.2 × 10^6/mL）. Total （13.4 ± 9.9 vs. 7.1 ± 6.8） and progressive （4.4 ± 3.9 vs. 2.2 ± 2.1） motility were significantly higher in subjects with lower scrotal temperatures. There was no statistical significant difference between electroejaculation and penile vibratory stimulation groups as regards any of the semen parameters. Subjects＇ age, infrequent ejaculation, injury duration and hormonal profile showed no significant effect on semen parameters. Conclusion： The defining characteristics of the seminogram in men with SCI are normal volume and count with decreased sperm motility, vitality and normal forms, and the increased number of pus cells. The most acceptable cause of the deterioration of semen is elevated scrotal temperature.     

Semen quality changes among 2343 healthy Slovenian men included in an IVF-ET programme from 1983 to 1996

To determine whether semen quality in Slovenians has changed over 14 years (1983-96), we analysed retrospectively the semen of 2343 healthy men with a normal spermiogram, who were partners of women with tubal infertility included in the IVF-ET programme. Age at semen collection, duration of sexual abstinence, semen volume, sperm concentration, total sperm count, percentage of spermatozoa with progressive motility, and normal morphology were determined. Multiple regression analysis was used to assess the changes in sperm characteristics according to the year of semen collection, year of the man's birth and the duration of sexual abstinence. Semen volume, sperm concentration, sperm count and total sperm motility did not change between 1983 and 1996, whereas between 1988 and 1996 rapid progressive sperm motility decreased by 0.95% per year (p < 0.0001). Semen volume, sperm concentration, and sperm count increased with duration of sexual abstinence. After adjustment for the year of semen collection and duration of sexual abstinence, multiple regression analysis showed that sperm concentration decreased by 0.67% per each successive year of birth (p = 0.03). Thus the sperm concentration decreased from 87.6 x 10(6)/mL in men born in the 1940s to 77.3 x 10(6)/mL in those born between 1956 and 1960. After 1960, sperm concentration was found to increase. In 2343 healthy men, no decline in semen quality, except in rapid progressive motility, was observed in the study period. Lower sperm concentration was found among men born between 1950 and 1960. This could be related to worse socio-economic status, stress or negative environmental factors in this time period.     

黄芪注射液对镉诱导大鼠精子畸形的拮抗作用

目的 :观察黄芪注射液拮抗氯化镉诱导的大鼠精子畸形作用。　方法 :将 30只SD雄性大鼠随机分成 5组 :低浓度黄芪组 (A1)、高浓度黄芪组 (A2 )、环磷酰胺组 (CP)、氯化镉组 (Cd)和对照组 (C)。预先连续 7d分别给A1组和A2 组大鼠腹腔黄芪注射液 5g/ (kg·d)和 10g/ (kg·d) ,Cd组和CP组大鼠分别腹腔注射等量蒸馏水作对照 ,之后连续 2 1dA1组、A2 组和Cd组大鼠分别同时腹腔注射氯化镉溶液 [0 .2mg/ (kg·d) ]。CP组大鼠给予 5 0mg/ (kg·d)腹腔注射染镉后第 2 2d处死大鼠 ,观察睾丸脏器系数、精子头计数、每日精子生成量、附睾尾精子计数和畸形率以及睾丸和附睾病理学。　结果 :A2 组睾丸脏器系数、睾丸精子头计数、每日精子生成量和附睾精子计数 [(5 .6 8±1.19)、(4 9.0 1± 8.78)× 10 6/g、(10 .2 5± 2 .30 )× 10 6/ (g·d)、(4 7.5 1± 2 2 .5 1)× 10 6/ml]明显高于Cd组 [(3.11± 0 .16 )、(37.5 9± 10 .6 3)× 10 6/g、(5 .31± 0 .32 )× 10 6/ (g·d)、(10 .89± 2 .4 5 )× 10 6/ml](P <0 .0 5或P <0 .0 1) ;A2 组大鼠精子畸形率 [(7.0 4± 0 .12 ) % ]明显下降 ,与Cd组 [(17.81± 1.5 5 ) % ]相比 ,差异有极显著性 (P <0 .0 1)。　结论 :黄芪注射液可拮抗氯化镉对生殖系统的损害作用 ,对防护     

Activity of testis angiotensin converting enzyme (ACE) in ejaculated human spermatozoa

Testicular angiotensin converting enzyme (ACE) isozyme is likely to play important functional roles in male reproduction. Several studies have shown that ACE is released from human spermatozoa during capacitation and that ACE is associated with reduced sperm motility. Recently, we established an assay to detect testicular ACE activity in human spermatozoa. The purpose of this study was to determine if testicular ACE activity is related to sperm motility in human ejaculates. Semen samples were collected from 80 infertile patients. According to the semen characteristics, they were divided into four (WHO) categories. Enzyme activities of ACE in spermatozoa (testicular ACE) and seminal plasma (somatic ACE) were spectrophotometrically determined. Total testicular ACE activity in spermatozoa was measured by solubilization of spermatozoa with Triton X-100. Membrane testicular ACE activity was measured in a sperm : PBS suspension. Sperm concentration and sperm motility were 136.6 +/- 154.1 x 10(6)/mL and 58.6 +/- 23.4%, respectively (mean +/- SD). Enzyme activities of membrane testicular ACE, total testicular ACE and somatic ACE were 0.273 +/- 1.219 microU/10(6) spermatozoa, 0.35 +/- 1.34 microU/10(6) spermatozoa and 684.7 +/- 226.6 mU/mL, respectively. A negative correlation was observed between sperm motility and membrane testicular ACE activity (p < 0.05). Membrane testicular ACE activity in 44 normal semen samples was 0.04 +/- 0.02 microU/10(6) spermatozoa, whilst that in 36 abnormal semen samples was 0.24 +/- 0.42 microU/10(6) spermatozoa. There was a significant difference between these two groups (p < 0.01). Membrane testicular ACE in sperm samples from normozoospermic men was significantly lower than that from oligoasthenozoospermic men (p < 0.05). These findings suggest that testicular ACE is released from normal functional spermatozoa for them to have fertilizing ability.     

Diurnal scrotal skin temperature and semen quality. The Danish First Pregnancy Planner Study Team

It is well established that heat is associated with reduced sperm production, but the role of physiological variation in temperature has never been scrutinized in humans. We studied diurnal scrotal temperature and markers of male fertility in a population of couples planning their first pregnancy. Sixty men from a cohort of couples who were planning their first pregnancy were included and scrotal skin temperature was monitored during 3 days using a portable data recorder. Working hours and working postures were recorded daily in a questionnaire. Each man provided a fresh semen sample and the couples were followed for six menstrual cycles or until a clinical pregnancy was recognized. The median value of scrotal skin temperature was 33.3 degrees C in the daytime and 34.8 degrees C at night. In periods of sedentary work, the median temperature was on average 0.7 degrees C higher (SE=0.2 degrees C). In addition, scrotal temperature was higher in the daytime, in summer, and in leisure time compared with working hours. Median sperm concentration among men with more than 75% of their daytime readings above 35 degrees C was 33.4 x 10(6)/mL, compared with 91.8 x 10(6)/mL for men with less than half of their readings above 35 degrees C (difference 58.4; 95% CI: 25.9-77.8 x 10(6)/mL). It is concluded that a sedentary position is a significant source of increased scrotal skin temperature, and even moderate and physiological elevation in scrotal skin temperature is associated with a substantially reduced sperm concentration. Sedentary work should be considered as an important potential confounder for reduced sperm count in epidemiological research.     

Treatment in male infertile clinic of Kaizuka Municipal Hospital

The results of treatment of 68 idiopathic male infertile cases are reported. The follow up period was 3 years and 8 months from the time the Department of Urology, Kaizuka Municipal Hospital had opened. The main treatments were human mammary gonadotropin-human chorionic gonadotropin (HMG-HCG) therapy and high ligation of left testicular vein in the cases accompanied with varicocele. Adjuvant therapy was administration of vitamin B12, herb medicine and antibiotic agents for prostatovesiculitis. Some cases were administered clomiphene citrate. The results in 43 cases at over 10 weeks after treatment was followed. Mean sperm count and mean sperm activity rate in 11 cases accompanied with no varicocele and whose sperm count was 2 approximately 45 x 10(6)/ml were raised from 11.0 +/- 8.0 SD x 10(6)/ml and 19.1 +/- 11.4 SD% to 22.7 +/- 16.8 SD x 10(6)/ml and 26.9 +/- 18.5 SD%, respectively after 20 weeks of HMG-HCG therapy. Efficacy was 7 out of 11 (63.6%) in sperm count and 6 out of 11 (54.5%) in sperm activity rate. Pregnancy was obtained in 3 cases. Effect of HMG-HCG therapy was not observed in 8 cases accompanied with varicocele and whose sperm count was 2 approximately 45 x 10(6)/ml. High ligation of left testicular vein was effective in all of the 4 cases accompanied with varicocele and whose sperm count was 2 approximately 45 x 10(6)/ml.(ABSTRACT TRUNCATED AT 250 WORDS)     

Preservation of testicular arteries during subinguinal microsurgical varicocelectomy: clinical considerations

Microsurgical varicocelectomy with intentional preservation of the testicular artery(ies) is regarded as the gold standard approach to varicocele repair. We sought to determine whether the number of testicular arteries preserved at the time of micro-surgical varicocelectomy predicts improvement in postoperative semen parameters. We analyzed the records of 334 infertile men who underwent varicocelectomy performed by a single surgeon using a subinguinal microsurgical technique between July 1996 and January 2003. We examined the association between the number of testicular arteries preserved at the time of varicocelectomy and serum follicle-stimulating hormone (FSH), luteinizing hormone (LH), varicocele grade, testicular volume, and postoperative improvement in semen parameters. Unilateral, left-sided varicocelectomy was performed in 194 men, while bilateral varicocelectomy was performed in 140 men. Mean (+/-SE) sperm concentration (20.1 +/- 1.5 x 10(6)/mL to 26.7 +/- 1.9 x 10(6)/mL, P =.001), percent motility (24.7 +/- 1.0% to 30.9 +/- 1.2%, P =.001), and percent normal morphology (35.8 +/- 1.4% to 37.7 +/- 1.5%, P =.046) improved significantly following varicocelectomy. The mean number of preserved testicular arteries was 1.5 on the left (range, 1-4) and 1.5 on the right (range, 1-4). The number of testicular arteries preserved at the time of varicocelectomy did not correlate significantly with preoperative assessment of serum FSH, LH, varicocele grade, and testicular volume or with postoperative improvement in semen parameters. Our data indicate that preoperative parameters are not predictive of the number of testicular arteries identified at the time of microsurgery. These data also suggest that the number of arteries identified and preserved with meticulous spermatic cord dissection does not correlate with improvement in semen parameters.     

Secular and seasonal changes in semen quality among young Danish men: a statistical analysis of semen samples from 1927 donor candidates during 1977-1995

The objective of this study was to investigate whether semen quality has changed during the years 1977-1995 in a group of unselected semen donor candidates, and to determine whether semen quality is subject to seasonal variation, by analysis of time- and season-related changes in semen quality using multiple regression and ANOVA. The study was based on analysis of the first semen sample delivered by 1927 semen donor candidates in Copenhagen during the period 1977-1995, with determination of semen volume, sperm concentration, total sperm count, percentage motile spermatozoa, and a semiquantitative sperm motility score. Multiple linear regression analysis with year, sexual abstinence and season as covariates showed a significant increase in mean sperm concentration from 53.0 x 10(6)/mL in 1977 to 72.7 x 10(6)/mL in 1995 (p < 0.0001) and in mean total sperm count from 166.0 x 10(6) to 227.6 x 10(6) (p < 0.0001). Mean semen volume and percentage motile spermatozoa did not change. Sperm motility deteriorated, as the spermatozoa in 74.2% of the samples were of excellent motility in 1977-1980 compared to only 41.9% in 1993-1995 (chi 2 = 130.0, p < 0.0001). Analysis of variance showed significant variation between seasons regarding sperm concentration (p < 0.0001) and total sperm count (p < 0.0001). Highest sperm counts were found in spring, with a mean concentration (95% C.I.) of 77.6 x 10(6)/mL (71.9-83.7), and lowest in summer, with a mean of 57.5 x 10(6)/mL (50.1-65.4). No other semen parameter varied with season. It is concluded that sperm counts increased, whereas sperm motility decreased, in a group of Danish semen donor candidates, from 1977 to 1995. Due to the retrospective design and the anonymity of the donors, we were unable to control for variation in donor age, and we cannot exclude the possibility that some donor candidates were selected by being accepted as donors by other semen donor services in Copenhagen. With these limitations in mind, we suggest our results should be interpreted cautiously and regarded as a contribution to the ongoing dispute on whether or not there is a continuous decrease in sperm quality. The seasonal variations found in sperm concentration and total sperm count were pronounced and were not attributable to seasonal differences in the length of sexual abstinence. Additionally, the same seasonal pattern was observed in five successive year-intervals. These findings strongly indicate that human testicular function is influenced by season, a phenomenon well known in many lower mammals.     

Evaluation of trend in semen analysis for 11 years in subjects attending a fertility clinic in India

Aim: The data on semen analysis of subjects attending the Fertility Clinic at NIHFW (National Institute of Health and Family Welfare) Munirka, New Delhi for the last 11 years were analyzed to verify the claims and speculations on declining sperm counts in men. Methods: Approximately 10 % of the records every year starting from 1990 to 2000 (numbering 1176 in total) were randomly selected for analysis. Subjects with azoospermia or severe oligozoospermia were excluded from analysis. Results: The average age of the men attending the infertility clinic was 31.2 years. The average semen volume and sperm count were found to be (2.6 ±0.1) mL and (60.6 ± 0.9) × 106/mL, respectively. No significant decline in sperm counts was observed in any year during the entire study period. Only 1.8 % of the total number of sperm counts in the random sampling were less then 20 × 106/mL. On the basis of WHO criteria on motility, the total percentage of non-progressive and non-motile sperm in the ejaculate was higher (63 %)     

Seminal vesicle aspiration in spinal cord injured men: insight into poor sperm quality

PURPOSE: In men with spinal cord injury poor quality semen is seen when performing electroejaculation and penile vibratory stimulation. We determined whether sperm stasis within the seminal vesicles is a potential cause of this problem. MATERIALS AND METHODS: Seminal vesicle aspiration was performed immediately before electroejaculation or penile vibratory stimulation in men with aspermia secondary to spinal cord injury. Sperm count and quality of seminal vesicle aspiration and subsequent ejaculation were compared with historical ejaculated counts, ultrasound findings and patient characteristics. RESULTS: Mean total number of right plus left seminal vesicle sperm plus or minus standard deviation was 511 +/- 960 x 10(6). Mean total number of sperm obtained by seminal vesicle plus electroejaculation or penile vibratory stimulation was 918 +/- 1,261 x 10(6). Average motility and viability of the seminal vesicle aspirated sperm were 1.3 and 3.2%, respectively. Average motility of the ejaculated sperm was 26.4% after seminal vesicle aspiration versus 16.3% in previous ejaculation induction procedures performed in the same patients. Seminal vesicle aspirated sperm represented 66% of the total number of sperm obtained during the session and was equal to 49% of the sperm obtained at previous electroejaculation or penile vibratory stimulation sessions. The period of abstinence correlated only with ejaculate count (simple regression p = 0.009). No other clinical characteristics had any effect on sperm count or quality. CONCLUSIONS: Large numbers of poor quality sperm are present within the seminal vesicles of spinal cord injured men and these sperm comprise a large portion of the specimens collected by electroejaculation or penile vibratory stimulation. This phenomenon is independent of the period of abstinence, implicating disordered storage of sperm due to spinal cord injury rather than infrequent ejaculation. The large number of senescent sperm within the seminal vesicles appears to be a primary cause of poor sperm quality in spinal cord injured men.     

Melatonin administration alters semen quality in healthy men

The role of melatonin in the regulation of reproduction in humans is unknown. We conducted a 6-month, double-blind, crossover study of a daily treatment dose of 3 mg melatonin or placebo given orally at 1700 hours in 8 healthy men. Semen quality (concentration, motility, and morphology), serum and seminal plasma 17-beta-estradiol (E(2)), testosterone, melatonin, and serum gonadotropin levels were determined every 3 months throughout the study. In 6 men, there was no change in semen quality or in serum and seminal plasma hormone levels during the study period. In 2 men, during the melatonin treatment period, sperm concentration decreased to 3 x 10(6)/mL and 12 x 10(6)/mL, and motility declined to 32% and 30%. These coincided with a decline in seminal plasma and serum E(2) levels and with an increase in testosterone:E(2) ratios. Six months after the cessation of melatonin, sperm concentration and motility were normal in 1 man but remained abnormal in the other one with a still elevated testosterone:E(2) ratio. Serum gonadotropin levels were unchanged during the study in all 8 men. Our preliminary observations suggest that long-term melatonin administration is associated with decreased semen quality in a number of healthy men, probably through the inhibition of aromatase at the testicular level.     

The mast cells in semen: their effects on sperm motility

This study was conducted to evaluate any possible association between mast cells and sperm concentration, morphology, and motility. The study comprised 400 patients who had applied for semen analysis. To evaluate mast cells, 6 smear slides were prepared for each subject and stained with 1% toluidin blue-pyronine (pH 4). The slides revealing any mast cells were labeled as mast+. Concentration and motility was evaluated through a Makler chamber. Kruger's strict criteria were used in morphometric analysis. The mean age of 86 mast+ cases (21% of total patients) was 31+/-6.7; progressive sperm motility rate was 33+/-21.2. The mean concentration was 32+/-30.2 x 10(6)/mL, and normal sperm percentage was 11.8+/-6.5. Progressive sperm motility rate in the mast- cases were 53+/-25. The mean age of mast cell+ patients was higher than that of mast cell- patients (t=3.57, p<.001), while they had lower sperm concentration (p>.05) and lower normal morphologic sperm rate (t=2.26, p<.024), compared to mast cell- patients. The relation between mast cell+ and mast cell- cases and sperm progressive motility was statistically significant (t=6.44, p<.001). It was concluded that sperm parameters were negatively affected by mast cells.     

Poor hypo-osmotic swelling test results from cryopreserved sperm despite preservation of sperm motility

Cryopreservation thawing of sperm leads to decreased motile density. Most donor programs select for cryopreservation a male with a high initial motile density and if the post-thaw semen has a motile density over 10 x 10(6)/ml, the specimen is thought to represent a fertile specimen. Some recent data suggest that males with normal motile densities but subnormal hypo-osmotic swelling (HOS) tests may be infertile. A study was thus performed to see if males with an adequate motile density after cryopreservation may still demonstrate membrane damage as evidenced by decreased viability and a poor HOS test. The semen species from seven men with motile densities greater than or equal to 10 x 10(6)/ml after freeze-thawing were evaluated for HOS changes and viability. Despite preservation of normal motile density, all 7 men had HOS scores below 50% following cryopreservation (though all were significantly above this level pre-freeze). The mean viability and HOS scores prefreezing were 70 +/- 9.7 and 68.5 +/- 9.5, and post thaw they dropped to 33.7 +/- 6.9 and 32.8 +/- 6.2. These data suggest a mechanism for impaired fertility even with adequate motile density of a thawed specimen that had been cryopreserved.     

Computer-assisted image analysis of sperm concentration in human semen before and after swim-up separation: comparison with assessment by haemocytometer

Evaluation of male fertility is based predominantly on results from semen analysis and determination of the sperm concentration is one of the main parameters of the analysis. The availability of a fully automated videomicrographic digital image analyser would offer both an objective and rapid method for determination of the sperm concentration. In the present study the sperm concentration in 327 semen samples was determined by haemocytometer according to the World Health Organization guidelines, and also by a computer-assisted digital image analyser system. Results were classified according to the routine procedure (haemocytometer) before statistical analyses. The computerized measurements caused a shift to the right in the frequency distribution of sperm concentration. Sperm concentrations were more often overestimated significantly (P less than 0.001) by the computerized measurements in semen samples with concentrations up to 80.0 x 10(6)/ml. This overestimation seemed to be caused by the presence of particles in seminal plasma that were recognized incorrectly as sperm by the computer program. The computerized digital image analyser gave an average sperm concentration of 2.2 +/- 0.6 x 10(6)/ml (mean +/- SEM) in 17 azoospermic semen samples while the routine procedure did not detect the presence of sperm cells. After removing the seminal plasma by washing and centrifugation with culture medium, and using the swim-up procedure to harvest motile sperm, the computerized measurements showed comparable results with the routine procedure for those sperm preparations (n = 44) with sperm concentrations greater than 5.0 x 10(6)/ml.(ABSTRACT TRUNCATED AT 250 WORDS)