Improvement in quality of cryopreserved human spermatozoa by swim-up before freezing

Selecting a population of spermatozoa by the swim-up technique yields, after freezing and thawing, a population of cells that contains proportionally more spermatozoa which are morphologically normal, fewer spermatozoa with damaged tail membranes, and a greater percentage of progressively motile spermatozoa with greater velocities and amplitudes of head displacement than those obtained after freezing and thawing the same semen samples in the normal way. This pattern was found for the semen from 10 patients and five volunteers. However, the cells selected by swim-up were as susceptible to the stresses caused by freezing and thawing as unselected spermatozoa in the original semen sample, and the improvement came from the better quality of the initial sample.     

Investigation of, the cause of low sperm motility in asthenozoospermic patients by multiple quantitative tests

Multiple tests were done on the ejaculates of 10 asthenozoospermic patients and nine healthy normozoospermic volunteers in an attempt to identify individually the cause of low sperm motility in these patients. Possible defects in the sperm plasma membrane and the motility apparatus of sperm, and in epididymal function affecting the development of motility, were investigated. The presence of seminal sperm antibodies or any motility-inhibiting factors in the seminal plasma that could be removed by washing were also tested. Each test was positive in only one or two patients but axonemal dysfunction was identified in nine patients. Removal of seminal plasma from asthenozoospermic samples did not improve sperm motility to any greater extent than with donor ejaculates, and the motile sperm of these patients exhibited characteristics mostly similar to those of donors under various incubation conditions. Selection procedures are, therefore, required to obtain samples of good quality sperm from such asthenozoospermic ejaculates.     

Testicular function in eight patients with seminoma after unilateral orchidectomy and radiotherapy

Testicular function was monitored in eight patients with low stage seminoma who were treated with radiotherapy following unilateral orchidectomy. The absorbed gonadal radiation dose ranged from 15 to 157.5 rad. At 10-24 months after radiotherapy, serum hormone levels, sperm analysis, sperm penetration into zona-free hamster ova (HOP-test) and lymphocyte chromosome abnormalities were evaluated. Two patients were azoospermic with elevated serum levels of LH and FSH. The remaining six patients had slightly decreased (n = 3) or normal (n = 3) seminal parameters. Their HOP rates were within the normal range. A low incidence of polyspermy (i.e. only one penetrating sperm per ovum) was found in the patients, suggesting low penetrability of motile sperm. A highly significant correlation was found between sperm count or sperm penetrability and time post-irradiation. The results indicate that restitution of testicular function is time-dependent.     

PULSATILE GnRH-THERAPY IN OLIGOZOOSPERMIC MEN DOES NOT IMPROVE SEMINAL PARAMETERS DESPITE DECREASED FSH LEVELS

In order to evaluate GnRH administration for the treatment of infertile men with elevated serum FSH levels we administered GnRH in pulses via portable electronic infusion pumps initially to seven patients with low sperm counts and high FSH values over 12 weeks and later to nine further patients over 24 weeks who also underwent testicular biopsies. Fifty microlitres containing 5 micrograms GnRH were infused subcutaneously for 1 min every 120 min in the short-term study and every 90 min in the long-term study. Although FSH levels could be lowered in both groups of patients, none showed any improvement in sperm count or other seminal parameters. Therefore, pulsatile GnRH treatment cannot be recommended for therapy of severe oligozoospermia with elevated FSH levels.     

True hermaphroditism with bilateral ovotestis: a case report

A true hermaphrodite with a 46 XX/47 XXY karyotype, gynaecomastia, hypospadia and scrotal gonads was investigated. Gonadectomy performed at 14 years of age revealed bilateral ovotestes. The ovarian portion contained follicles of all developmental stages. The testicular portion was immature consisting of seminiferous cords with Sertoli cells at various steps of differentiation and few germ cells within massive aggregates of collagenous connective tissue. Leydig cells as well as germ cells remained in an embryonic stage of development. Sections of a differentiated Wolffian duct (ductuli efferentes, epididymis, vas deferens) as well as of a Müllerian duct (hypoplastic fallopian tube) were found adjacent to both gonads. Postoperative treatment consisted of androgen substitution therapy leading to progression of puberty.     

Immuno-competent cells in the murine epididymis

Cryostat sections of epididymides from mice were stained with monoclonal antibodies against immuno-competent cells. This investigation was undertaken to gain basic data about the distribution of macrophages. T lymphocytes, MHC class II and MIF in the normal murine epididymis to establish the mouse as a model for immunological epididymal research. The most important findings were as follows. (1) Macrophages, T lymphocytes, MHC class II and MIF positive cells were distributed similarly in the caput, corpus and cauda epididymis. (2) Macrophages were the most frequent leucocytes and the majority were located in the peritubular layer. (3) The MHC class II determinant was also expressed mostly in the peritubular layer and interstitium. These cells were similar in appearance and location to macrophages. (4) Significantly fewer T lymphocytes were found and their main location was the interstitium. T-helper and T-suppressor/cytotoxic lymphocytes did not differ significantly in their regional or histological distribution patterns. (5) The ratio of T-helper to T-suppressor/cytotoxic lymphocytes was 1:1. (6) MIF was detected almost exclusively in blood vessels and the surrounding connective tissue. (7) No invasion of leucocytes into the epididymal lumen was observed. It is concluded that macrophages seem to be the most important immunological cell type in the murine epididymis.     

Subnormal sperm parameters in conventional semen analysis are associated with discrepancies between fertilization and pregnancy rates in in-vitro fertilization and embryo transfer

Three hundred and twenty-eight consecutive treatment cycles in 168 couples were analysed retrospectively in order to examine the influence of conventional semen analysis results on the outcome of in-vitro fertilization and embryo transfer with respect to the occurrence of both fertilizations and pregnancies. All treatments were performed under maximally standardized and controlled conditions. Each of the three main determinants of the spermiogram, namely the concentration, motility and morphology of sperm in seminal plasma, was of significant importance for fertilization and subsequent pregnancy. Best correlations were achieved by counting the number of progressively (a+b) motile sperm and the number of normally formed sperm in seminal plasma. The pregnancy rate was reduced significantly in cases in which the sperm concentration was < 10 x 10(6) ml-1 (P < 0.01), or in which there was < 40% progressively motile sperm (P < 0.001), or < 30% normally formed sperm (P < 0.001). If more than one parameter in the spermiogram was abnormal, the fertilization rate depended mainly on the most disturbed sperm parameter. The implantation rate as well as the pregnancy rate was reduced significantly in patients with low progressive sperm motility and normal morphology rates. The difference could only be attributed partially to the lower number of embryos replaced. In conclusion, subnormal sperm quality seems to interfere with developmental stages beyond the process of fertilization.     

The influence of inflammation of the human male genital tract on secretion of the seminal markers α-glucosidase, glycerophosphocholine, carnitine, fructose and citric acid

Biochemical analysis was made of specific accessory gland products in the ejaculates of 362 men suffering from various acute inflammatory diseases of the reproductive tract and 33 normozoospermic patients acting as controls. The ejaculate content of the epididymal markers alpha-glucosidase and L-carnitine, but not glycerophosphocholine, was significantly reduced in ejaculates from men with epididymitis; citric acid was reduced in men suffering from prostatitis; both citric acid and alpha-glucosidase were reduced in men suffering from adnexitis. The ejaculate content of epididymal and prostatic markers in prostato-urethritis (adnexitis), where the exact localization of the inflammation was unclear, was not as low as in epididymitis or prostatitis. Seminal vesicle function, as judged from semen volumes and seminal fructose, was not different in these groups of patients. The results, although strongly related to the clinical diagnosis, were unrelated to the microbiological flora of the semen and indicate that both the epididymis and the prostate glands are involved in some forms of adnexitis.     

Diurnal variations in scrotal temperature of normal men and patients with varicocele before and after treatment

Scrotal temperature was monitored using a portable data recorder for periods of 24 h in six normal volunteers and 48 infertile patients with unilateral varicocele while subjects pursued their regular daily activities. Temperatures during sleep ( T _s) were generally higher than daytime values ( T _D), probably as a consequence of thermal insulation in bed. These diurnal variations were found to be less pronounced in the infertile patients than in volunteers (T_s- T_D= 0.29°C ± 0.06°C us 0.88°C ± 0.12°C;P<0.01). Moreover, scrotal temperatures at night of patients and volunteers were indistinguishable statistically, but were different during daytime hours. After successful ligation or embolization of the spermatic vein in 16 patients, no change in scrotal temperature was observed. Although sperm counts were higher after treatment, this difference was not significant (67.4 ± 17.2 × 10⁶ vs 105.8 ± 25.5 ± 10⁶; P>0.05). The data support the view that varicocele-related damage to the testis results from a lack of adequate cooling, and that treatment does not normalize the temperature pattern.