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E Strehler

Publications and source records attributed to E Strehler.

At least 37 records · Page 2Linked to original sources

Submicroscopic mathematical evaluation of spermatozoa in assisted reproduction. I. Intracytoplasmic sperm injection. (Notulae seminologicae 6).

After a large introduction concerning the influence of sperm quality in the success of in vitro fertilization, the debated problem of the importance of the sperm quality in intracytoplasmic sperm injection is investigated. The spermatozoa are studied by electron microscopy, and the results evaluated by the formula of Baccetti et al. (1995). The quality of spermatozoa has been correlated with the success of ICSI. The first conclusion has been that ejaculates which produced embryos contained higher percentages and overall higher total number of 'healthy' spermatozoa than those which obtained fertilizations arrested at the two pronuclei stage, or showed no fecundation at all. A second conclusion was that the quality of sperm organelles is mainly involved in the oocyte activation, but after the 2 pronuclei stage, segmentation usually proceeds with few influences from sperm quality. The highest involvement of spermatozoa is therefore the initiation of oocyte activation. A third conclusion confirmed the need of ultrastructural evaluation of sperm cell organization, deeply investigating the inner organelles. Moreover, we observed that the head organelles are more involved that the tail ones. Three acrosomal characteristics are concerned: the shape, the dimensions and the content. All of them are significantly better in the oocyte activating than in non activating spermatozoa. Also the nuclear shape and the status of the chromatin (frequently strictly interdependent) are significantly concerned with the oocyte activation and with the embryo segmentation. In the tail, the mitochondrial shape is significantly different in activating and not activating spermatozoa, and also in those producing embryo segmentation. Moreover, the absence of dynein arms seems to be determinant in impeding the oocyte activation.

Fertilization in Vitro↗

[Electron microscopic analysis of possible ultrastructural damage to spermatozoa after preparation by various methods].

We investigated the possibility of ultrastructural damage to human spermatozoa induced by different sperm preparation techniques. Ejaculates from 20 normozoospermic men were divided into equal aliquots and processed by glass wool filtration, Percoll density gradient centrifugation, and a simple two step centrifugation procedure which served as a control. Ultrastructural damage was assessed by scanning electron microscopy. Compared with Percoll density gradient centrifugation, glass wool filtration yielded a significantly increased proportion of intact acrosomes (p < 0.05). However, both preparations gave significantly better results than control (p < 0.001 and p < 0.01 for glass wool filtration and Percoll centrifugation, respectively). The conspicuous bending of sperm tails after Percoll centrifugation has a yet unknown importance. In conclusion, both glass wool filtration and Percoll centrifugation are efficient techniques for the accumulation of spermatozoa with intact acrosomes. Because of the higher sperm recovery rate, glass wool filtration appears to be the more appropriate method.

Acrosome↗

In vitro kallikrein treatment of human spermatozoa: stimulation of sperm penetration into zona-free hamster eggs depends on the quality of initial semen parameters but not on constituents of the kallikrein-kinin system in seminal plasma.

We investigated the effect of an in vitro kallikrein treatment of human spermatozoa on their ability to penetrate zona-free hamster eggs. Sixty-four infertility patients were classified into four groups (normo-, oligo-, astheno-, and oligoasthenozoospermia) in order to find out which men would benefit most from this procedure. An aliquot of each ejaculate was diluted with Ham's F-10 medium supplemented with human serum albumin and kept at 22C for 1 hour. Another aliquot was incubated with kallikrein (5 KU/ml.) for one hour at 22C. These sperm suspensions were then washed twice, subjected to a swim-up preparation, and subsequently incubated at 37C for 2 hours. The mean penetration rates of kallikrein-treated samples were significantly higher than those of the corresponding control assays in all but the oligoasthenozoospermia group. Our results suggest a direct proteolytic action of kallikrein because the stimulation of penetration into hamster eggs was not correlated with the amount of constituents of the kallikrein-kinin system (kinins, kininogen, kininase) in seminal plasma.

Animals↗

[Do preparation techniques induce ultrastructural sperm damage? An electron microscopic study].

We investigated the possibility of ultrastructural damage to human spermatozoa induced by different sperm preparation techniques. Ejaculates from 20 normozoospermic men were divided into equal aliquots and processed by glass wool filtration, Percoll density gradient centrifugation, and a simple two-step centrifugation procedure which served as a control. Ultrastructural damage was assessed by scanning electron microscopy. Compared with Percoll density gradient centrifugation, glass wool filtration yielded a significantly increased proportion of intact acrosomes (p < 0.05). However, both preparations gave significantly better results than control (p < 0.001 and p < 0.01 for glass wool filtration and Percoll centrifugation, respectively). The significance of the conspicuous bending of sperm tails after Percoll centrifugation is not yet known. In conclusion, both glass wool filtration and Percoll centrifugation are efficient techniques for the accumulation of spermatozoa with intact acrosomes. Because of the higher sperm recovery rate, glass wool filtration appears to be the more appropriate method.

Acrosome↗

[Correlation between cytogenetic anomalies of human spermatozoa and sperm morphology as well as age of patients studied].

Sperm chromosomes from 15 fertile men were analyzed after fusion of their spermatozoa with zona-free hamster eggs. The total proportion of abnormal metaphases as well as the proportions of aneuploidy and structural aberrations were calculated for every man and examined for linear correlations with sperm morphology and the age of the persons studied. A positive correlation between the cytogenetic parameters and the percentage of abnormal sperm morphology was not evident, suggesting that assessment of sperm morphology cannot be used as an indicator of chromosomal damage in human spermatozoa. In contrast, there was a more distinct positive correlation between the age of donors and the three cytogenetic parameters studied. Analogous calculations for two other study groups (group S: male subfertility; group HA: habitual abortions of the wives with unknown etiology) provided less clear results that may be attributed to the lower number of patients or to the existence of special indications as mentioned above.

Abortion, Habitual↗

Cytogenetics of human spermatozoa: correlations with sperm morphology and age of fertile men.

Sperm chromosomes from 15 fertile men were analyzed after fusion of their spermatozoa with zona-free hamster eggs. The total proportion of abnormal metaphases as well as the proportions of aneuploidy and structural aberrations were calculated for every man and examined for linear correlations with [1] sperm morphology and [2] the age of the persons studied. A positive correlation between the cytogenetic parameters and the percentage of abnormal sperm morphology was not evident, suggesting that assessment of sperm morphology cannot be used as an indicator of chromosomal damage in human spermatozoa. In contrast, there was a more distinct positive correlation between the age of donors and the three cytogenetic parameters studied.

Adult↗

[New perspectives of in vitro kallikrein treatment. II. Study of the hamster ovum penetration test in relation to the kallikrein-kinin system].

A significant enhancement of the fertilizing capacity of human spermatozoa by Kallikrein has recently been described in a former study. To assess the therapeutic possibilities it would be of great interest to know more about the biochemical effect of Kallikrein on the fertilizing capacity of human spermatozoa. This fact made the authors investigate 64 semen samples in which the binding score and the fertilizing capacity were determined after application of Kallikrein in a HOP-test (hamsterovum-penetration-test) and compared to the control. Furthermore these results were compared to the varying concentrations of the components of the Kallikrein-Kinin-system. An application of Kallikrein in the swim-up preparation resulted in a higher motility in patients with asthenozoospermia, whereas the patients belonging to the group of oligoasthenozoospermia showed a lower progressive motility compared to the control. The binding score (number of affixed spermatozoa) showed nearly the results in both preparations. On the other hand the fertilizing capacity in the Kallikrein preparation was significantly higher. A possible connection with the concentrations of the components of seminal plasma has not been found, yet, but regarding all results a direct enzymatic effect of Kallikrein on fertilizing potential can be assumed.

Animals↗

[New perspectives of in vitro kallikrein treatment of human spermatozoa. I. Study of the hamster ovum penetration test in relation to different andrologic patient groups].

Whereas the enhancement of motility by Kallikrein is described several times, the effect on fertilizing capacity of human spermatozoa has not been investigated sufficiently yet. This fact induced the authors to start a further investigation on 64 patients by the means of zona-free hamster oocytes. The patients were differentiated by the parameters density and motility. The sample was divided up into two aliquots. Differing from the control the other aliquot was prepared with Kallikrein (5 KU/ml) before swim-up and preincubation for one hour at 22 degrees C. Comparing the results we found looking at the whole collective (n = 64) a significant enhancement of the fertilizing capacity. 34 of 64 patients showed an improvement, only 4 patients deteriorated. While the groups normo-, oligo-, and asthenozoospermia reached a higher fertilizing capacity under Kallikrein-treatment (probability of error 2.5%), a Kallikrein application in case of oligo-asthenozoospermia had no significant influence on fertilizing capacity. A view of the whole results leads to the assumption of a direct effect of Kallikrein and to the possibility of use for GIFT and IVF procedure.

Adult↗

Differentiation of the atrioventricular node, the atrioventricular bundle and the bundle branches in the bovine heart: an immunohistochemical and enzyme histochemical study.

The previous observations of differences between different cardiac regions (ventricular myocardium, atrial myocardium, Purkinje fibre system) with respect to the maturation of the M-line region and the establishment of mature metabolic characteristics, have been extended. It was found that M-line maturation proceeds differently also between different regions of the conduction system. The M-line proteins, myomesin and MM-creatine kinase, were detected earlier, by means of immunohistochemistry, in the AV bundle and bundle branch cells than in the AV node cells. Also, a difference was observed in large foetuses. Striations in the AV node were less evident than in the AV bundle and the bundle branches in sections incubated with antibodies against myomesin as well as against MM-creatine kinase. Using enzyme histochemistry it was observed that the differences in metabolic properties between the AV node, the AV bundle and the bundle branches on the one hand, and the ordinary myocardium on the other, of adult hearts, are not established at the early stages. No clear difference in activity of succinate dehydrogenase was seen between the conduction tissues and the ordinary myocardium in the foetal hearts, while the conduction tissues showed a lower activity in the adult hearts. Furthermore, the pattern of activity of mitochondrial glycerol-3-phosphate dehydrogenase between the conduction tissues and the atrial and ventricular myocardium was quite different in early foetal stages compared with the adult stage.

Animals↗

Differentiation of Purkinje fibres and ordinary ventricular and atrial myocytes in the bovine heart: an immuno- and enzyme histochemical study.

The differentiation of Purkinje fibres and ordinary ventricular and atrial myocytes in bovine hearts was studied with specific antibodies against M-line proteins (MM-creatine kinase and myomesin) and with enzyme histochemistry (succinate dehydrogenase and mitochondrial glycerol-3-phosphate dehydrogenase). MM-creatine kinase was detected at an earlier stage in Purkinje fibres and atrial myocytes than in ordinary ventricular myocytes. The findings are in agreement with previous ultrastructural observations that an earlier appearance of a dense M-band occurs in Purkinje fibres than in ordinary ventricular myocytes. Myomesin was detected in all three cell types even at early foetal stages, in accordance with suggestions that it is an integral component of the myofibrillar structure. The activity of succinate dehydrogenase gradually increased in both ordinary ventricular and atrial myocytes, while the activity of mitochondrial glycerol-3-phosphate dehydrogenase was high at different stages of early foetal development in the two tissues, finally becoming low in the adult stage. The activity of succinate dehydrogenase and mitochondrial glycerol-3-phosphate dehydrogenase seemed to remain unchanged in the Purkinje fibres from early to late foetal stages. The present study shows that the Purkinje fibres are already different from ordinary ventricular myocytes at early foetal stages and that the two cell types differentiate in different ways. It is concluded that there are also developmental differences between ordinary ventricular and atrial myocytes.

Animals↗

Differentiation of the myofibrils and the intermediate filament system during postnatal development of the rat heart.

The differentiation of the myofibrils and the intermediate filament system during postnatal development of the rat heart has been investigated. Several aspects of some of the structural proteins, that means the intermediate filament subunit skeletin, myosin, and the myofibrillar M-line proteins MM-creatine kinase and myomesin have been studied by using gel electrophoresis as well as enzyme and immunohistochemical techniques in combination with electron microscopy of both plastic and cryosectional material. We show that marked changes take place in the organization of the intermediate filament system and in the contractile apparatus, both in atria and in ventricles of the rat heart during postnatal development. In the newborn rats no dense myofibrillar M-bands were present in the M-region and the sarcomeric bands were irregular while in the four-week-old rats dense M-bands composed of a set of five crossbridges interconnecting the thick filaments were present. The sarcomeric bands were now regular. These observations are related to the presence of different isomyosins in the atria and in the ventricles of the newborn and the four-week-old rats, to the observation that MM-creatine kinase was only present in the M-region in the four-week old rats and to the physiological maturation of the heart.

Animals↗

Chromatin decondensation of human sperm in vitro and its relation to fertilization rate after ICSI.

The inability of sperm chromatin to decondense has been implicated in the failure of fertilization, This study was undertaken to identify the relationship between sperm chromatin decondensation in vitro after incubation with follicular fluid at various points in time and fertilization or pregnancy rates after intracytoplasmic sperm injection. Moreover, an attempt was made to determine whether this test could be used as a predictive test for the outcome of ICSI. Thirty-two infertile couples undergoing ICSI therapy were included in this prospective study. One milliter of semen from each sample was mixed with 1 mL of follicular fluid obtained from ICSI patients at the time of oocyte retrieval and incubated for 24 h. Many smears were made directly after semen liquefaction at the following time intervals: 30, 60, and 120 min and 24 h. Chromatin decondensation was evaluated with acridine orange staining. The mean percentage of uncondensed chromatin of spermatozoa in the native semen samples was 25 +/- 18.3%, which increased within 24 h to 91 +/- 9.5%. On the other hand, the fertilization and ongoing pregnancy rates were 64 +/- 21.7% and 20%, respectively. However, no correlations were found between chromatin decondensation at various point of time (30, 60, and 120 min and 24 h) and fertilization rate. No correlation was shown between the chromatin decondensation and sperm counts in the ejaculate. morphology, or the percentage of condensed chromatin. In light of this study, chromatin decondensation in vitro cannot be recommended for predicting the fertilization potential of spermatozoa and pregnancy rates in the ICSI program. Further research is necessary, especially in cases where ICSI is being considered as a therapeutic option.

Adult↗