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[Cold preservation of human sperm].

The authors report the development and investigation of a new technique for freezing and storage of human spermatozoa. Utilizing isolated container of different kind the average rate of freezing was between 1,3 grd/min and 14 grd/min in the sphere from 5 degrees C to--10 degrees C and between 2,9 grd/min to 55 grd/min in the sphere from--10 degrees C to--60 degrees C. The technical expense was low. Semen specime were provided by 33 andrological patients. A total of 53 samples of human semen were frozen and stored for periods of time up to 25 days. The authors used the cryoprotective medium from Behrman and Sawada. The best results were obtained in the mode of freezing at 2,8 grd/min respectively 12 grd/min in ejaculates of original motility about 80%. The recovery of motility and fertilizability of spermatozoa was 50% of original motility, in cases of oligospermia only 34%. The results of this new technique of freezing and storage of human spermatozoa are similar to other new reports in literature.

Freezing

Human sperm antigens and antisperm antibodies. II. Age-related incidence of antisperm antibodies.

Naturally occurring human antisperm antibodies (ASA) detectable by immunofluorescence, were found to have a peak indicence of 90% in both sexes before puberty. Thereafter, the incidence declined to about 60% and persisted through life. This age-related incidence resembled that for foreign and not self antigens. The natural incidence of the six antibodies under investigation varied: the most frequent were antibodies to acrosomal antigen Acl and equatorial antigen; the least frequent were antibodies to acrosomal antigen Ac2 and sperm nuclear protamine; and between these were antibodies to the mainpiece of tail and post-acrosomal region. Irrespective of their natural incidence, these antibodies increased at a comparable rate in men following vasectomy indicating these six sperm antigens have comparable immunogenicity. These results provide evidence for the lack of immunologic tolerance in man toward many sperm antigens. The variation in the natural incidence of individual ASA is explanable by differences in prevalence of crossreaction between each sperm antigen with exogenous antigens. Thus, antibodies to acrosomal antigen Acl and equatorial antigen, which occurred most frequently, appeared to crossreact with ubiquitous microorganisms.

Adolescent

Elimination of errors induced during a routine human sperm motility analysis.

Influence of some variables on the accuracy of sperm motility evaluation was studied by analyzing normal seminal specimens with the aid of themultiple exposure photography (MEP) method. Results of this study demonstrated the existence of variations in sperm motility between view areas of the same sample or between different samples of the same specimen. Variations were found even when the same view area was analyzed intermittently during a short period of time. Motility was practically unaffected when a regular sized drop was kept within the preparation for about 20 min. The light from the microscope neither stimulated nor depressed sperm motility when they were illuminated continuously for that period of time. Percent of motility and sperm velocity was directly related to the thickness of the examined drop when the latter ranged between zero and ten micrometers. According to these findings it is suggested that several view areas from various drops of the same specimen should be analyzed with each view area being inspected several times before motility of that field is assessed. Motility should be evaluated from drops of standard and constant thickness; otherwise major errors in sperm motility assessment are to be expected.

Diagnostic Errors

Juvenile diabetes and human sperm quality.

Spermiological findings in 25 juvenile diabetics in the age range 16 to 22 years (average 18.5) have been compared with normal spermiological findings in 24 individuals of the same age (16 to 22 years, average 18.7). A drift to lower sperm values in the group of the juvenile diabetics became evident in all parameters observed. As concerns motility and morphology, the differences between the groups reached statistically significant values.

Adolescent

A comparison of subjective measurements of human sperm motility and viability with two live-dead staining techniques.

Two commonly used live-dead stains (eosin-nigrosin (EN) and eosin-opal blue (EOB)) were compared with the estimated active spermatozoa in semen samples from patients attending an infertility service. Twenty-eight semen samples were analyzed throughout the day of their collection by estimating the number of active spermatozoa and by staining a portion of the incubated sample (37 degrees C) with each stain. The samples were analyzed 30, 60, 120, 240, and 360 minutes after the initial collection. At 30 minutes there were no significant differences between the estimated values and those of either stain. The slope of the EN stain closely paralleled that of the estimated measurements throughout the remainder of the time periods, while the EOB slope was somewhat steeper. Repeatability of values for semen samples obtained on different days was generally good for each stain. A comparison of semen samples from 85 patients, 30 minutes after collection, showed no significant differences between the numbers of estimated active sperm and the percentage live using the EN stain. Studies of 132 semen samples using the EN stain showed a positive correlation with the over-all quality of sperm motility. The results indicate that there is a role for live-dead staining in assessing semen quality.

Cell Biology

Autoimmune human sperm antibodies and age in males.

The incidence of autoimmunization to spermatozoa, investigated by means of the Kibrick technique, increased with age in 913 males unselected with regard to fertility. By contrast, the incidence of sperm agglutinin-postive sera decreased with age in 1600 male partners in sterile couples. The increasing incidence of autoimmunization to spermatozoa offers one explanation of the lowering of fertility with increasing age, but sperm antibodies seem to be more important as a cause of sterility at younger ages. The presence of spermatozoa may not be necessary for immunization since sperm agglutinins were found in a 9-year-old boy.

Adolescent