The objective of this study was to develop a relatively simple test to evaluate the functional integrity of the membranes of human spermatozoa. As in some other species, human spermatozoa 'swell' under hypo-osmotic conditions due to the influx of water and the expansion of the membranes. A mixture of equal parts of fructose and sodium citrate (150 mosmol) with calculated ionic strength of 0.15 resulted in a maximal number of clearly identifiable swollen spermatozoa. Only small variations were seen when different aliquants of the same semen samples were separately evaluated. A high correlation (r = 0.94) was obtained between expected and observed values of swollen spermatozoa when known amounts of heat-treated spermatozoa, unable to undergo swelling, were added to untreated spermatozoa. A good correlation (r = 0.90) was also observed between the % spermatozoa in a semen sample that were capable of undergoing swelling and the % of denuded hamster oocytes that were penetrated by capacitated spermatozoa from the same semen sample. By contrast, the correlations between % sperm swelling in ejaculates and % normal sperm forms, % motile spermatozoa and % spermatozoa that do not stain with eosin-Y (supravital stain) in the same ejaculates were 0.30, 0.61 and 0.52, respectively. Therefore, the hypoosmotic swelling technique to evaluate the functional integrity of the sperm membrane appears to give high repeatability and accuracy and is closely correlated to the in-vitro fertilizing ability of spermatozoa. It may be a useful addition to the standard semen analysis.
Human ejaculates (n = 83) were analyzed for standard sperm parameters (concentration, motility, and morphology), as well as for the ability of the spermatozoa to react (swell) in a hypoosmotic medium (Jeyendran et al, 1984). Subsequently, the fertilizing capacity of the spermatozoa was tested by their ability to fertilize human oocytes in vitro. Although the sperm concentration was adjusted for in vitro fertilization, no adjustments were made for sperm motility and morphology. Correlation of the in vitro fertilizing capacity of the spermatozoa with the hypoosmotic swelling test (r = 0.56) was much higher than with standard sperm parameters (r varied from −0.04 to 0.25). Complete overlap was noted with standard semen parameters whether the ejaculate did or did not fertilize oocytes and ranged from very low to very high values in both cases. By contrast, all the semen samples that fertilized oocytes showed a 60% or higher reaction in the hypoosmotic swelling test, whereas the majority of the “infertile” semen samples showed less than 60% swelling. It therefore appears that, under the conditions of our studies, the hypoosmotic swelling test is a more accurate predictor of successful in vitro fertilization outcome than the conventional semen parameters. A combination of all parameters, however, is likely to be most useful. The hypoosmotic swelling test is simple and economical, and it is recommended that this test be further scrutinized for its value as an additional tool in the assessment of the in vivo fertilizing capacity of ejaculated spermatozoa.
Stained sperm morphology slides from an in-vitro fertilization programme were evaluated by the modified Williams system, the World Health Organization (WHO) system and morphometric size measurements (image analysis) to determine which system(s) were better predictors of fertility. Amorphous sperm head shape was the best predictor of fertility in both the modified Williams system and the WHO system, predicting the proper fertility status in 65% of the cases. Using only normal sperm in the statistical model, normal sperm became a correct predictor in 66% of the cases in the modified Williams system. Morphometric measurements were not useful in predicting the fertility status of these samples. These studies indicate that morphology can not be used as a sole predictor of fertilizing capacity of human spermatozoa.
Blood and semen samples obtained from 24 "normal" volunteers were analyzed for 16 different biochemical parameters. These included: Ca, Mg, K, Na, Zn, C1, P, glycerolphosphorylcholine (GPC), carnitine, fructose, uric acid, prostatic acid phosphatase, alkaline phosphatase, glutamic oxaloacetic transaminase (SGOT), lactic dehydrogenase (LDH), and glutamic pyruvate transaminase (SGPT). With the exception of uric acid, all the biochemical constituents in seminal plasma were either significantly higher (p < 0.001, except alkaline phosphatase which was significant at p < 0.05) or significantly lower (p < 0.001) than in blood serum. Further, potassium (r = 0.51); carnitine (r = 0.54); and SGOT (r = 0.70) showed a significant direct relationship in blood and seminal plasma. Vergleich biochemischer Parameter vom menschlichen Blutserum und SeminalplasmaZusammenfassung: Blut-und Samenproben von 24 ,,normalen" Freiwilligen wurden nach 16 verschiedenen biochemischen Parametern analysiert. Diese beinhalten: Ca, Mg, K, Na, Zu, C1, P, Glycerolphosphorylcholine (GPC), Carnitin, Fructose, Harnsaure, saure Prostataphosphatase, allealische Phosphotase, Glutamat-Oxalat-Transaminase (SGOT), Laktat-Dehydrogenase (LDH) und Glutamat-Pyruvat-Transaminase (SGPT). Mit Ausnahme von Harnsaure sind alle biochemischen Bestandteile im Seminal-Plasma entweder signifikant hoher (p < 0.001, mit Ausnahme der alkalischen Phosphatase, die bei p < 0.05 signifikant war) oder signifikant niedriger (p < 0.001) als im Blutserum. Ferner zeigten Potassium ( y = 0.51); Carnitin ( y = 0.54); und SGOT / y 0.70) eine signifikante direkte Beziehung zwischen Blut-und Seminal-Plasma.
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