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For the evaluation of sperm functions it is important to assess the acrosome reaction after induction with various stimuli. Acrosome reaction tests normally include the capacitation of spermatozoa, treatment with an inducer, and detection of acrosomal loss by dyes, lectins or antibodies. Since most of these methods are time-consuming or require expensive equipment, paramagnetic beads coated with Pisum sativum agglutinin (PSA) were investigated for their usefulness in facilitating the detection of human sperm acrosome reaction. Binding of PSA beads to the acrosomal region increased significantly after incubation of capacitated spermatozoa with 10 microM A23187 (20.3 +/- 6.7% [mean +/- SD, absolute binding], n = 21), 1 mM dibutyryladenosine cyclic monophosphate (17.1 +/- 8.5%, n = 25) and 10 mM phorbol myristate acetate (21.1 +/- 12.5%, n = 10). Bead binding was significantly reduced by pre-incubation with a protein kinase inhibitor. Beads bound to Concanavalin A (ConA) were also attached to the acrosomal region after induction of the acrosome reaction by A23187 or dbcAMP, but a lower number of spermatozoa were bound to ConA-beads than to PSA beads. Pre-treatment of spermatozoa with alpha-methyl-D-mannoside before addition of the PSA beads markedly decreased bead binding, which indicates its mannose-specificity. Electron microscopic examinations demonstrated that PSA beads mainly bound to membrane structures of spermatozoa that were undergoing, but had not completed the acrosome reaction.

Microsurgical epididymal sperm aspiration (MESA) and testicular sperm extraction (TESE) require close cooperation between andrologists, urologists and gynaecologists. Intracytoplasmic sperm injections were established in Giessen in March 1994 and embryo transfer (ET) was performed in 342 of 375 patients (91.2%). The percentage of pregnancies and ongoing pregnancies are 35.4% per ET (32.3% per cycle) and 25.1% per ET (22.9% per cycle), respectively. Microsurgical procedures such as epididymovasostomy or vasovasostomy and cryopreservation of human semen are also established methods. The purpose of the present study was to describe the andrological work-up for patients before MESA and TESE. Experiments demonstrate that incubation of testicular tissue samples in IVF medium and treatment with 1 mg ml-1 pentoxifylline increase the number of extracted motile spermatozoa. Centrifugation of the medium results in a further concentration of sperm cells. If no motile spermatozoa can be found in the supernatant medium, they may be extracted directly from the testicular tissue samples by means of a micromanipulator.

The reasons for the discrepancy between 'patency' and 'pregnancy' in the outcome of microsurgical refertilization are partially unknown. The quality of the intra-operative aspirate and of the spermatozoa at the level of the anastomosis are discussed worldwide as important factors influencing the success of fertilization in the case of post-operative patency. In 152 men undergoing microsurgical refertilization (vasovasostomy, tubulovasostomy, microsurgical epididymal sperm aspiration, transurethral resection of the ejaculatory duct), 281 aspirates were classified intra-operatively according to Silber, and post-operatively using the Shorr staining technique. In 62 aspirates a computer-aided sperm analysis (CASA) was performed. The percentage of intact spermatozoa decreased from 94.9% at the caput to 9.4% at the cauda epididymidis. The post-operative classification demonstrated an acceptable correlation (0.71) to all grades of intraoperative classification. There was a good correspondence in Silber 4 and 5 but worse in Silber 1 and 2. In CASA, the percentage of motile spermatozoa was not different between epididymal and post-epididymal aspirates. Furthermore, velocity parameters did not differ significantly, but there was a significantly higher straightness of post-epididymal spermatozoa in comparison to epididymal spermatozoa.

Smoking and varicocele are frequent findings in the medical history and physical examination of patients attending andrological 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 = 30) and varicocele patients (n = 30) compared with normal fertile donors (n = 20). The acrosome reaction was detected by triple staining after 3 h of incubation at 37 degrees C, followed by treatment with 0.1% dimethylsulphoxide (spontaneous acrosome reaction) and 10 microM calcium ionophore A23187 (induced acrosome reaction) for 1 h at 37 degrees C. Acrosin activity was measured by gelatinolysis. The diameters around the sperm heads after gelatinolysis and the percentages of spermatozoa showing halo 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 microns, P < 0.001) were significantly lower in the varicocele group than in the samples 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.

Human spermatozoa from healthy donors (n = 7) were washed in Tyrode's medium containing 0.6 mg/ml freeze-dried egg yolk, filtered through glass wool and exposed to 0.5, 5, 50, 500 or 1000 microM hydrogen hexachloroplatinate. Viability, membrane integrity and the acrosome reaction were examined using trypan blue exclusion, hypo-osmotic swelling test and fluoresceinated Pisum sativum agglutinin, respectively, after incubation for 3 or 6 h at 37 degrees C. While sperm motility, viability and membrane integrity were not affected by the platinum compound after incubation for 3 h, the number of acrosome-reacted spermatozoa increased from 16.0 +/- 6.4% (control, mean +/- SEM) to 21.0 +/- 3.3 (0.5 microM), 22.3 +/- 4.3% (5 microM), 28.0 +/- 4.3% (50 microM, p < 0.01), 29.3 +/- 3.9% (500 microM, p < 0.01) and 43.9 +/- 7.4% (1 mM, p < 0.001); a further increase was detected after incubation for 6 h. However, the percentages of dead and immotile spermatozoa and those with defective membranes were also higher, suggesting that the acrosome reaction was caused by degenerative processes after long-term incubation. In conclusion, hydrogen hexachloroplatinate does not affect sperm motility, membrane integrity or viability, but it does induce the acrosome reaction after incubation for 3 h before cytotoxic effects are measurable. Similar effects of halide salts of platinum on receptor-mediated exocytosis have been described in other cells such as mast cells and basophils in vivo and in vitro.

Tail stump syndrome, which may be associated with primary ciliary dyskinesia, is also associated with morphological defects of the flagellum resulting in severe asthenozoospermia. Until recently, these morphological anomalies caused definite male infertility. Today, however, new methods such as micromanipulation techniques provide a rational therapy for this patient group. A pregnancy followed by living offspring was achieved following the intracytoplasmic injection of immotile spermatozoa from a patient with tail stump syndrome.

The present study examines the release of angiotensin-converting enzyme (ACE) from human spermatozoa during capacitation conditions and in correlation to acrosome reaction and cell death. The ACE content of spermatozoa was measured by treating the cells with different detergents. Glass wool-filtered and washed human spermatozoa were incubated for 3 hours at 37 degrees C. Percentages of acrosome-reacted and dead spermatozoa did not change significantly, but the ACE release increased from 0 to 2.93 +/- 0.44 mU/100 x 10(6) spermatozoa after 180 minutes (P < 0.001). In order to study the influence of acrosome reaction on ACE release, the acrosome reaction of noncapacitated spermatozoa was induced by 10 microM calcium ionophore A23187. The percentages of acrosome reaction and viability in noncapacitated spermatozoa as well as the ACE release were compared to corresponding data from experiments using capacitated spermatozoa (3 hours, 37 degrees C) from the same donors. Although the number of living acrosome-reacted spermatozoa after ionophore treatment (30.5 +/- 4.0%) was significantly higher than after capacitation (13.3 +/- 2.8%, P < 0.001), ACE release from ionophore-treated, noncapacitated spermatozoa was lower (P < 0.05). The data indicate that ACE release from human spermatozoa during capacitation is independent of acrosome reaction. Measurement of ACE release may be a clinically useful assay for human sperm capacitation.

The present study was performed to determine the usefulness of the alpha-sympathomimetic midodrin for diagnosis and treatment of functional sperm transport disturbances. 140 andrological patients consulting due to severe oligozoospermia, hypospermia or partial/complete retrograde ejaculation were included. Ejaculates were examined 30 min after intravenous injection of 5-15 mg midodrin. Sperm concentration, motility and alpha-glucosidase as a marker of both epididymal function and sufficient passage through the vasa deferentia and the ejaculatory duct were measured and the values compared to those in the ejaculates obtained before therapy. In 23 of 140 patients, sperm concentration or total sperm count was improved by more than 10 million spermatozoa ml-1 or 20 million spermatozoa per ejaculation. Alpha-glucosidase in the seminal plasma increased in two cases. The present study demonstrates that severe oligozoospermia may be caused by functional transport disturbance of semen from the epididymis to the efferent duct system. In these cases, midodrin is of diagnostic and therapeutic value; however, it should be emphasized that its effectiveness cannot be predicted individually.

In view of demographic trends, andrology will increasingly be concerned with problems of the aging male. These will involve aspects of fertility and general quality of life rather than specific clinical pictures. While there is considerable interindividual variation, fertility in men usually persists well into old age. However, increased occurrence of spontaneous mutations means a higher genetic risk for children of older fathers. Diminished androgen levels in old age lead to diminished sexual activity and changes in muscles and bones, which raise the question of hormone substitution.

The storage of spermatozoa from tumour patients in sperm banks is an important medical task. As a result of improved treatment strategies the survival rate, particularly in patients with testicular tumours and Hodgkin's disease, is excellent if the diagnosis is made early. However, the necessary therapeutic measures often lead to the mutilation of reproductive function. At the Department of Dermatology at the University of Munich a sperm bank has been in existence since 1974; most of the patients who consult the sperm bank are men with testicular tumours. A synopsis of the last 12 years provides information about the patients, indications for storage, the method of preservation used and the fate of the cryopreserved samples used for insemination. To guarantee that optimal results are achieved, cryopreserved sperm samples should only be used by gynaecologists who are particularly experienced in the treatment of sterility.