Peroxidative damage induced by reactive oxygen species (ROS) has been proposed as one of the major causes of defective sperm function. In previous studies of the production of ROS in semen, the contribution of contaminating leucocytes was not assessed. We determined the levels of ROS in 60 semen samples from men attending our infertility clinic and demonstrated by performing extraction experiments with antibody-coated magnetic beads that, within this unselected population of patients, leucocytes were the major source of ROS in the low-density Percoll fraction. Of the sperm motion parameters examined using computerized semen analysis, beat-cross frequency was the only one significantly affected by the ROS in semen.
Human gamete interaction is of fundamental biological importance, yet the molecular interactions between spermatozoa and the zona pellucida are poorly understood. Surprisingly, the role of the polypeptide backbone of zona pellucida glycoprotein 3 (ZP3), the putative ligand for spermatozoa activation, has been largely overlooked. Purified recombinant human ZP3 was expressed in Escherichia coli as a C-terminal fusion to the dimeric glutathione S-transferase (GST) from Schistosoma japonicum and was shown to induce acrosomal exocytosis in live, capacitated human spermatozoa. The level of exocytosis is comparable with that obtained using purified, glycosylated, recombinant human ZP3 [van Duin, M., Polman, J. E. M., DeBreet, I. T. M., Van Ginneken, K., Bunschoten, H., Grootenhuis, A., Brindle, J. and Aitken, R. J. (1994). Biol Reprod. 51, 607-617]. These data imply that the polypeptide chain of human ZP3 contributes to recognition of spermatozoa during acrosomal exocytosis in vitro.
Monoclonal antibodies were used to detect seminal leucocytes in 19 men with urethritis. A wide range in the number and type of leucocytes between individuals was documented (median 2.06 x 10(6)/ml, range 0.6-29.89 x 10(6)/ml); 22% of the men had less than 1 x 10(6) leucocytes/ml. The results suggest that the threshold of greater than or equal to 1 x 10(6)/ml proposed by the World Health Organisation to indicate genital tract infection is unsuitable for men with urethritis.
This review addresses critical issues in the selection of semen donors who are very fertile. Traditional semen parameters have been employed and are still used to assess pre- and post-freeze samples in order to discriminate between donors of high and low fecundity. The most predictive factor is the number of motile spermatozoa per straw and the number of motile spermatozoa inseminated. Nevertheless, no absolute standards for fertile samples can be derived from the basic semen examination. The employment of sperm function testing such as the hamster penetration test or computerized motility analysis has been shown to enhance moderately the prediction of fertility of semen samples however, further studies are necessary to determine if these improvements are clinically useful. The need to determine with a high degree of confidence the fecundity of donor semen is enhanced by limitations in the number of pregnancies allowed per donor. The recent publication of league tables in the UK has put extra pressure in clinics to use highly fertile donors. Spermatozoa are also cryostored for patients prior to cancer treatment. With the development of intracytoplasmic sperm injection every sample produced by cancer patients can be stored irrespective of the quality. However, several factors need to be elucidated to maximize the fertility of those patients. The establishment of regional centres in Europe will be a good starting point to deal with many of the issues raised in this review.
The relationship between asymptomatic urethral infection and seminal white blood cells, as detected using the peroxidase enzyme system, was examined. Eighty-four semen donors were tested. Twenty-four (29%) were diagnosed as having an active urethral genital infection. There was no statistical relationship between the total number of concentrations of peroxidase-positive cells and a urethral genital infection. Further studies should concentrate on the subtypes of seminal leucocytes and their surface receptors using monoclonal antibodies.
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