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Biomedical subjects

H J Glander

Publications and source records attributed to H J Glander.

At least 37 records · Page 2Linked to original sources
PubMed1996

Low expression of adhesion molecules and matrix proteins in patients showing poor penetration in zona-free hamster oocytes.

The expression of adhesion molecules and matrix proteins by human spermatozoa as well as their binding to and penetration of zona-free hamster eggs were investigated in 17 patients by means of flow cytometry. Both binding and penetration of hamster oocytes and expression of alpha- and beta-chains of beta 1, beta 3 and beta 4 integrins were determined before and after induction of the acrosome reaction. The expression of the integrin ligands, fibronectin and laminin were also analysed. Significant differences in the expression of very late antigen (VLA) integrins, VLA alpha 4-chain (CD49d), the classical fibronectin receptor VLA alpha 5-chain (CD49e), leukocyte function-associated molecule-3 (LFA-3; CD58) and fibronectin were observed between patients showing good (> 10%) or poor (< 10%) penetration in the sperm penetration assay (P = 0.0068). It is concluded that these adhesion molecules are intimately involved in the sperm-oolemma interaction. Since no differences were observed in either spermiogram parameters or the percentages of acrosome-reacted spermatozoa in the two groups, sperm-oolemma binding and penetration should be regarded as discrete parameters of sperm function.

Animals↗
PubMed1996

Detection of mRNA transcripts of beta 1 integrins in ejaculated human spermatozoa by nested reverse transcription-polymerase chain reaction.

Matrix proteins (e.g. fibronectin, laminin and vitronectin) and their receptors, especially beta 1 integrins, may play a role in sperm-oocyte interactions. These proteins are well described on spermatogenic cells but especially in ejaculated spermatozoa, the presence of mRNA as a necessity for their synthesis is not clear. Therefore the mRNA synthesis of the beta 1, alpha 3, alpha 4, alpha 5 and alpha 6 chains (fibronectin and laminin receptors) of the beta 1 integrins was investigated by nested reverse transcription-polymerase chain reaction (RT-PCR) in spermatozoa of human semen samples (n = 18) which showed normal classical semen parameters after swim-up preparation. After the second amplification, we obtained specific signals for mRNA transcripts of the beta 1, alpha 3, alpha 4 and alpha 6 chains in 17 of 18 semen samples investigated. All control PCRs were negative for somatic cell transcripts and contamination by somatic cells was excluded. These results demonstrate for the first time that human ejaculated spermatozoa contain the mRNA transcripts of beta 1 integrins and support the role of these proteins in adhesive mechanisms in the fertilization process.

Actins↗
PubMed1996

Insulin-like growth factor-I and alpha 2-macroglobulin in seminal plasma correlate with semen quality.

Insulin-like growth factor-I (IGF-I) and alpha 2-macroglobulin (alpha 2-M) are believed to be involved in the development of germ cells. IGF-I is mainly controlled by concentrations of human growth hormone (HGH), influences cell proliferation and differentiation and its action is mediated by insulin-like growth factor-binding proteins (IGFBP), placental protein 14 (PP14) and prostate-specific antigen (PSA). alpha 2-M acts as a broad spectrum proteinase inhibitor and a binding protein for many cytokines and hormones, e.g. inhibin and activin. This study was designed to identify concentrations of these molecules in seminal plasma in normal semen samples of healthy men, correlations with semen quality, the relationship of IGF-I and alpha 2-M with factors affecting male fertility, and whether vasectomy influences the concentrations of these molecules. Concentrations of IGF-I and alpha 2-M in human seminal plasma were related to semen quality, basal concentrations of HGH, testosterone, IGFBP-3, soluble fibronectin receptor (sFNR), PSA and PP14 in seminal plasma and to serum concentrations of luteinizing hormone (LH) and follicle stimulating hormone (FSH). Commercially available assays were used to analyse 69 semen samples of various quality and 11 post-vasectomy samples. IGF-I concentrations in seminal plasma were significantly correlated with the percentage of morphologically normal spermatozoa (r = 0.748, P = 0.00001) and sperm concentration (r = 0.301, P = 0.011), but negatively correlated with serum FSH (r = -0.506, P = 0.00006) and PSA in seminal plasma (r = -0.388, P = 0.00009). Total alpha 2-M was significantly correlated with sperm count (r = 0.423, P = 0.0005), percentage of progressively motile spermatozoa (r = 0.444, P = 0.00019), quality of motility (sperm motile efficiency, r = 802, P = 0.00001) and straight line velocity (r = 0.411, P = 0.0013). Correlation between the sperm concentration and HGH in seminal plasma was weak (r = 0.287, P = 0.015). Vasectomy reduced the concentration of total alpha 2-M (P = 0.00008) and HGH (P = 0.0068) in the seminal plasma; IGF-I was also reduced after vasectomy when the total ejaculate amount was considered. Thus IGF-I and alpha 2-M are significant for the germ cell development: IGF-I in the maturation of spermatozoa and alpha 2-M in progressive motility.

Follicle Stimulating Hormone↗
PubMed1994

Influence of the epididymis on integrins and matrix proteins of human spermatozoa.

Spermatozoa alter their antigenity and develop the ability of binding to zona pellucida and to vitellus during the epididymal transit. The involvement of spermatozoal adhesion molecules (AM), especially beta 1-integrins in this binding process between the gametes is very likely. Therefore, the influence of epididymis on the binding capacity of AM was indirectly investigated by comparison of AM on ejaculated and on testicular human spermatozoa and spermatogenic cells, respectively. The AM-expression was detected on ejaculated spermatozoa by flow cytometry (EPICS PROFILE II, Coulter Corp., Krefeld, Germany) and on spermatogenic cells by immunohistology using monoclonal antibodies against. beta 1-integrins and matrix proteins. The alpha-chains 3, 5, 6 of the beta 1-integrins and fibronectin were found on ejaculated spermatozoa as well as on spermatogenic cells except spermatogonia. The alpha-4-chain and laminin could only be detected after the epididymal transit of spermatozoa, whereas, the beta-chain of beta 1-integrins was missing on the ejaculated spermatozoa. These findings suggest that AM are predominantly expressed on spermatogenic cells before the epididymal transit but an epididymis-mediated modulation of AM, i.e. masking, demasking or activating of cryptodeterminants is possible. On this way dysfunction of epididymis could be followed by an impairment of AM-mediated binding capacity of spermatozoa.

Antigens, CD↗
PubMed1994

Spermiologic investigations before and after retrograde sclerotherapy of varicocele (preliminary communication).

A selected patient group (n = 17) with left-side varicocele was treated by retrograde sclerotherapy. Before and about six months after the treatment of varicocele the classical spermiogram, a computer aided sperm motion analysis, the acrosomal intactness, the maturity of nuclear proteins and the DNA-normality of the spermatozoa were examined. The sclerotherapy led to an increase of sperm concentration (P < 0.05), a significant improvement of sperm movement quality (P < 0.01; sperm motile efficiency and velocity straight linear), a significant increase of percentage of spermatozoa non-stainable by aniline-blue and with green fluorescence in the acridine orange test (P < 0.05). It can be concluded that the varicocele-treatment causes improvements of sperm-qualities which are not detected by the classical spermiogram.

Acrosome↗
PubMed1993

Evidence of beta 1 integrins and fibronectin on spermatogenic cells in human testis.

Interactions between human spermatozoa and oocytes are an essential event in the process of fertilization. Cell-cell and cell-matrix interactions in somatic cells are mediated by adhesion molecules such as beta 1 integrins (very late antigens; VLA). Therefore, we investigated the expression of beta 1 integrins and the matrix proteins collagen IV, fibronectin and laminin in human testis by immunohistology. Monoclonal antibodies against the beta chain of beta 1 integrins reacted with the basement membrane of the tubuli seminiferi, spermatocytes, spermatids and testicular spermatozoa. The alpha 3, -5 and -6 chains of beta 1 integrins showed the same pattern, whereas, the alpha 1, -2 and -4 chains could not be detected on spermatogenic cells. These VLA subunits were localized on endothelial cells, leukocytes and basement membranes. Matrix proteins such as laminin, collagen IV and fibronectin were detectable as components of basement membranes in human testis. Germinal cells except spermatogonia expressed fibronectin only. These results demonstrate that beta 1 integrins and matrix proteins in human testis are normally expressed on somatic tissue and that germinal cells, especially spermatocytes, spermatids and spermatozoa show positive reactions with antibodies against the VLA-3, -5 and -6 complexes and fibronectin. These findings suggest a production of beta 1 integrins and fibronectin during the spermatogenesis and a role of these proteins in adhesive mechanisms of spermatozoa similar to somatic cell-cell or cell-matrix interactions.

Adult↗
PubMed1993

Beta 1-integrins of spermatozoa: a flow cytophotometric analysis.

The expression of adhesion molecules on human ejaculated spermatozoa was investigated by means of flow cytophotometry using monoclonal antibodies against eight different adhesion molecules. Spermatozoa from infertility patients who exhibited normal semen parameters showed specific fluorescence after incubation with antibodies against very late antigens VLA alpha 4, VLA alpha 5, VLA alpha 6, and corresponding matrix proteins laminin and fibronectin. Semen samples from men with teratozoospermia or oligoasthenoteratozoospermia were characterized by reduced expression of adhesion molecules. Normal and pathological semen samples differed mostly in the expression of VLA alpha 5. After artificial induction of the acrosome reaction by a cold shock method and incubation with 2 mM Ca2+, a changed pattern of adhesion molecules on the sperm surface was found for VLA alpha 6 as the most prominent molecule. After treatment of spermatozoa with 0.1% Triton X-100, the number of cells with adhesion molecules increased significantly. This result suggests that adhesion molecules are an integral part of sperm structure.

Acrosome↗
PubMed1993

Semen samples with teratozoospermia show a lower percentage of spermatozoa with detectable adhesion molecules (AM).

Adhesion molecules (AM) are cell surface proteins which interact with ligands and mediate cell-cell bindings. Possibly, the AM are involved in the spermatozoal adhesion to oocytes. Human spermatozoa showed several adhesion molecules on their surface: the alpha chains of the beta 1-integrins VLA alpha 4 (CDw 49d), VLA alpha 5 (CDw 49e) and VLA alpha 6 (CDw 49f), the beta-chain of beta 2-integrins (CD 18) and the matrix proteins laminin and fibronectin. Semen samples with severe teratozoospermia were characterized by a significantly lower percentage (P < 0.01) of spermatozoa with VLA alpha 4, VLA alpha 5 and laminin. VLA alpha 5 (the alpha-chain of the fibronectin receptor) showed the most significant difference between the "normal" and teratozoospermic semen sample groups. This phenomenon could contribute to the explanation of a lower fertilizing capacity of teratozoospermic semen samples.

Cell Adhesion Molecules↗
PubMed1990

[A simple method for separating fresh and cryopreserved human sperm using dextran-visotrast density gradient centrifugation].

A procedure for the separation of fresh and cryopreserved human spermatozoa is presented, compared with the Ficoll method according to Harrison [7] and clinically tested. The principle includes a filtration through glass wool and a gradient centrifugation. The gradient medium consists of Ca2(+)-free buffer, Dextran "70" and Visotrast, a radio-opaque medium. The sperm concentration, the sperm-migration-index and the sperm morphology checked by FACS 440 corresponds to the results of the Ficoll-method described by Harrison [7]. Furthermore, our method is usable for semen samples after adding of a cryoprotective medium with egg yolk and glycerol. An improvement of the chance of conception after artificial insemination by husband (AIH) by our method could not be found up to now.

Centrifugation, Density Gradient↗
PubMed1990

[Fertility assessment based on sperm examination].

Male fertility is indirectly evaluated by analysis of semen. Two questions must be answered in connection with an investigation program. 1. Which prerequisites must the spermatozoa fulfill to the fertilization of oocytes in vivo and to the induction of a normal embryogenesis? 2. Which laboratory tests can be taken as an in vitro model for the fertilization process? Fertilization consists of various steps which require different spermatozoal functions. Several methods were developed to test in vitro whether the semen sample is capable to realize the sequence of fertilization, whereas, it is supposed that there is a close connection between the laboratory test in vitro and the fertilization in vivo. The methods of sperm analysis are presented and their medical practice and predictive value is discussed.

Female↗
PubMed1990

[Development of partner relations and of the child after successful therapeutic donor insemination].

The psychological development of couples and children was examined after successful therapeutic insemination by donor (TID). 222 couples were investigated by sending questionnaires to the families. 122 (55%) couples fulfilled our requests. The answers suggested that there was no deviation from the behaviour of the control groups concerning attitudes towards pregnancy and delivery, actual interrelation between wife and husband, degree of neurosis of the parents and children. The conclusion could be drawn that there was no need for more restriction on the TID provided that a good psychological diagnostic regarding the special TID-problems was performed before the insemination treatment.

Female↗
PubMed1989

[Expression of leukocyte antigens in human sperm].

The expression of leukocyte antigens on human ejaculated spermatozoa and lymphocytes was compared by means of the immunofluorescence technique and monoclonal antibodies against anti-HLA class I (pan-HLA-A,B,C) and class II (DR, DR/DP). Lymphocytes showed specific fluorescence phenomena after incubation with the antibodies. However, in the case of spermatozoa no immunofluorescent labeling was detected. Modifications of the method of sperm preparation (Ficoll centrifugation, swim-up technique, addition of azide, fetal calf serum or benzamidinchloride) did not lead to different results. We were not able to find HLA-antigen expression on fresh human spermatozoa using immunofluorescence.

Epitopes↗
PubMed1989

[Function of the spermatozoal plasma membrane in oocyte fertilization. Cell electrophoresis and immunofluorescence microscopy studies].

The influence of the capacitation on the electric charge of the spermatozoal surface and the unspecific spermatozoon-oocyte-binding are not fully understood up to now. Therefore, human semen samples were divided into two subgroups of spermatozoa by the "swim up" method. Both of the subgroups were examined by means of the micro-electrophoresis and the immunofluorescence technique using antihuman fibronectin antibodies. Cellular electrophoresis: The 2 subgroups of spermatozoa prepared of one semen sample showed a significantly different electrophoretic mobility (EPM) after washing and resuspension in an isotonic glycin-HCl-buffer (pH 2.0). In contrast, after resuspension in Baker's-buffer (pH 7.4) no difference between the 2 subgroups of one semen sample was detectable. The incubation in a medium with Ca++ caused secondly alterations of the EPM with the result of a smaller difference between the two subgroups of spermatozoa. Albumin also modified the EPM of spermatozoa. Immunofluorescence technique: 69 per cent of spermatozoa with a normal shape and only 17 per cent of them exhibiting a pathological shape showed a positive fibronectin-fluorescent-band on the equatorial segment called equatorial fibronectin band (EFB). Spermatozoa separated by "swim up" from semen samples with various semen parameters showed different percentages of spermatozoa with EFB. There was a significant difference between the spermatozoa of the two subgroups after "swim up" concerning EFB.

Electrophoresis↗
PubMed1989

[Partnership status in preparation for therapeutic donor insemination].

An investigation is presented which inquiries the psychological problems of sterile couples after diagnosing of male infertility. The interpretation of conversation topics accomplished by 460 couples suggests that coping with infertility by the couple, the character of the desire for a child, the level of stress tolerance, the ability to cooperation and the agreement to compromise determine the psychological situation of a couple infertile by male. These facts should be taken into consideration in regard of the therapeutic insemination by donor.

Adaptation, Psychological↗
PubMed1988

[Multiple pregnancy following in vitro fertilization and embryo transfer].

In a 32 year old patient 14 eggs were harvested and inseminated with donor semen. Six embryos were transferred resulting in a triplet-pregnancy. In spite of a septic disease three preterm but healthy children were born by caesarean section 199 days after ET (male 1620.0; female 1180.0; male 1560.0). Up to nowadays there is not any handicap. We don't know a general but safe method for avoiding multiple pregnancies.

Adult↗
PubMed1988

[Gossypol and other nonhormonal substances for the inhibition of male fertility].

Several international articles dealing with 3 substances, which act as a male contraceptive agent, were reviewed. The drugs gossypol, alpha-chlorohydrin and 6-chloro-6-deoxyglucose caused a reversible infertility without disturbances of sexual potency, but their toxic side effects exclude a clinical application at present.

Contraceptive Agents, Male↗