ISSN:0303-4569    

DOI:10.1111/j.1439-0272.1996.tb02749.x

出版商:Blackwell Publishing Ltd    

年代:1996

数据来源: WILEY

摘要:

Summary.Two methods of sperm preparation forin vitrofertilization were compared: the swim‐up technique vs. the migration‐sedimentation technique. The study comprised fresh semen samples obtained from 25 couples treated in theIn VitroFertilization Unit. Oocytes aspirated in a single cycle were divided into two groups, each inseminated by sperm prepared by one of these techniques. Motility, degree of motility, and normal morphology were improved by both methods. The improvement was greater when the migration‐sedimentation technique was applied. However, fertilization rate was significantly higher after the swim‐up technique. In order to clarify this contradiction, an additional group of 26 semen samples was divided and then prepared by the swim‐up or migration‐sedimentation techniques. Sperm quality was examined up to 72 h after separation. Compared with the swim‐up technique, sperm characteristics were better after separation by the migration‐sedimentation technique. However, this difference abated after 24 h. The better results of the swim‐up technique in the ‘survival experiment’ may explain its improved performance inin vitrofertilization, despite lower separation capacity. Thus, the migration‐sedimentation technique is not recommended for sperm preparation inin vitrofertilization.Migration‐sedi

ISSN:0303-4569    

DOI:10.1111/j.1439-0272.1996.tb02750.x

出版商:Blackwell Publishing Ltd    

年代:1996

数据来源: WILEY

摘要:

Summary.In order to ensure fertility, mammalian spermatozoa have to undergo acrosome reaction, the most obvious morphological change during this being the exposure of the inner acrosomal membrane. In the present study, the acrosome‐reacted human spermatozoa were successfully separated without loss of viability by using cell affinity chromatography on Concanavalin A (Con A) Sepharose. Con A demonstrated affinity for both the intact and the acrosome‐reacted spermatozoa regardless of their viability; the latter, however, gave higher affinity than the former against Con A. Prior to the column chromatography, the immotile spermatozoa and the seminal plasma were excluded by means of a modified swim‐down procedure and the resulting spermatozoa were subsequently immobilized by slow rate cooling in ice‐cold water. Cell affinity chromatography was performed at 4 °C. To prevent mechanical trapping of the spermatozoa among the packed gel beads, the column was interconnected with a reservoir, the vertical drive of which was allowed to lose the gel bed and thereby release the trapped spermatozoa. Stepwise competitive elution with 5.0 μM mannose and 25% heat‐inactivated human serum was capable of separating the intact spermatozoa and the acrosome‐reacted spermatozoa from each other. The acrosome reaction rate of sperm fraction which was adsorbed to Con A Sepharose and eluted with 25% serum was found to be 83±2.3%, and motility and viability of these fractions were measured to be 80±6.3% and 83±7.6%, respectively (n= 8, mean±SD). The status of the acrosome in a final preparation (motility 92%, acrosome reaction rate 88%) was observed by scanning electron microscopy, and 81% spermatozoa lost their acrosome cap.Acrosome‐

ISSN:0303-4569    

DOI:10.1111/j.1439-0272.1996.tb02751.x

出版商:Blackwell Publishing Ltd    

年代:1996

数据来源: WILEY

摘要:

Summary.The aims of this study were to compare thein vitroeffects of 3.6 mM and 7.2 mM pentoxifylline on the ability of spermatozoa to generate reactive oxygen species (ROS) and on lipid peroxidation (LPO). Semen samples were obtained from 10 asthenozoospermic men who had been previously identified as producing ROS after addition of Phorbol 12‐myristate 13‐acetate (PMA) during the screening of patients attending with male factor infertility. Spermatozoa were prepared by a swim‐up technique from unprocessed semen and divided into 3 aliquots. To the control aliquot [A] an equal volume of BWW medium was added. To aliquots B and C an equal volume of BWW medium containing pentoxifylline was added to obtain final concentrations of 3.6 and 7.2 mM, respectively. ROS production was measured from peak luminescence (mV 10−7sperm) using a lucigenin chemiluminescent probe. LPO was also measured in the medium surrounding the spermatozoa after 30 min exposure to pentoxifylline using the thiobarbituric acid (TBA) assay for malondialdehyde (MDA). The reduction in ROS production was significantly greater in the samples exposed to 7.2 mM pentoxifylline as compared with the control and 3.6 mM pentoxifylline samples. There was no significant difference in peak luminescence between control and 3.6 mM pentoxifylline specimens. Both concentrations of pentoxifylline caused comparable reductions in MDA concentration in the medium (P<0.05) surrounding the spermatozoa compared with control after 30 min exposure. Extracellular ROS generation may damage surrounding healthy spermatozoa. These findings suggest that higher concentrations of pentoxifylline are protective against ROS release in susceptible spermatozoa and may also reduce collate

ISSN:0303-4569    

DOI:10.1111/j.1439-0272.1996.tb02752.x

出版商:Blackwell Publishing Ltd    

年代:1996

数据来源: WILEY

摘要:

Summary.Sperm cell plasma membrane and the outer acrosomal membrane fuse profusely during the acrosome reaction. The process is triggered by extracellular signals that elicit several intracellular events leading ultimately to membrane fusion. We have developed a streptolysin O permeabilizing protocol that selectively affects the spermatozoon plasma membrane without causing a significant loss of the acrosomal content. Most of the acrosomal acid phosphatase remained sperm‐associated even after a 20 min incubation at 37°C. However, the presence of 100 μM Ca2+in the incubation buffer stimulates the release of the enzyme. The reaction was followed biochemically, measuring the acid phosphatase activity released to the medium and morphologically by the binding of fluorescein isothiocynate‐conjugated peanut agglutinin and by electron microscopy. The results show that the streptolysin O permeabilized spermatozoon is a promising model for studying the complex set of events mediating and regulating the acrosome reaction.Sperma

ISSN:0303-4569    

DOI:10.1111/j.1439-0272.1996.tb02753.x

出版商:Blackwell Publishing Ltd    

年代:1996

数据来源: WILEY

摘要:

Summary.The fixation of testicular tissue with glutardialdehyde destroys the antigenicity of cell epitopes in many cases. To obtain both, morphological and immunohistochemical examination, a fixation which could preserve the antigenicity and condition of the testicular structure was sought. The solution obtained was a mixture of 3.7% formalin with 0.2% glutardialdehyde and 0.05% saponin in a phosphate buffer of pH 7.4. Blocks of human testes were immersed in this fixative, embedded in Epon 812 or paraffin for conventional light and electron microscopy and immunohistochemical staining. The immunohistochemical examination was focused on the lamina propria of the human seminiferous tubules. Good preservation of the structure was observed both in light and electron microscopy. Cytological details were seen by light microscopy, especially the recognition of single tumour cells. Electron microscopically, all cells of the seminiferous tubules and the lamina propria showed a well‐preserved internal structure. At the same time the peritubular cells of the lamina propria exhibited a well‐expressed vimentin and desmin immunoreactivity, thus providing evidence that the corresponding epitopes retain their antigenicity under these fixation conditions. The applied fixation procedure provides comparable results in preservation of the structure to glutardialdehyde but does not destroy the antigenicity of epitopes.Testicular tis

ISSN:0303-4569    

DOI:10.1111/j.1439-0272.1996.tb02754.x

出版商:Blackwell Publishing Ltd    

年代:1996

数据来源: WILEY

摘要:

Summary.Thein vitrobinding capacity of spermatozoal integrins to matrix components after disintegration of sperm membranes was evaluated. The percentage of spermatozoa with functionally‐relevant integrins was determined before and after devitalization of spermatozoa, which were resuspended in seminal plasma or in culture medium. The devitalization was performed by cryoshock or by incubation of spermatozoa with triton X–100 in a concentration ranging from 0.01 to 1.0%. The spermatozoal integrins were detected by the binding of anti‐integrin antibodies and flow cytometry and the functional activity was monitored by the binding of the spermatozoa to the matrix components in a cell attachment assay. The seminal plasma decreased the binding of anti‐integrin antibodies to the spermatozoal surface and the binding of spermatozoa to ligands and matrix components, respectively. In contrast, the expression of fibronectin and laminin on spermatozoa was increased. Not all spermatozoa, which expressed integrins on their surface bound to the ligands in the cell attachment assay. These results suggest that the detectable integrins only partially exert functional relevance. It can be concluded that the spermatozoa with fragile plasma membranes are more prone to functional inactivation of their integrins by the seminal plasma.Sper

ISSN:0303-4569    

DOI:10.1111/j.1439-0272.1996.tb02755.x

出版商:Blackwell Publishing Ltd    

年代:1996

数据来源: WILEY

摘要:

Summary.Male Wistar rats were fed diets with different Mg content, ranging from 70 to 850 ppm Mg, for 30 days. After 0, 10, 20 and 30 days, some of the rats were sacrificed for measuring weight, lipid peroxidation, Fe, vitamin E, Na+, K+, Mg2+and Ca2+content of testes. After 30 days, the morphology of the testes was investigated by electron microscopy. Mg deficiency induced an increase in weight, Na+, Ca2+and Fe content and a reduction of K+ and Mg2+content. Vitamin E content was reduced and the content of malondialdehyde as an indicator of lipid peroxidation was increased. Mg deficiency induced morphological alterations in up to 40% of the spermatids in the 70 ppm Mg group, which consisted: 1) in injured stretching of spermatids; 2) in an irregular arrangement of coarse fibres with missing microtubulus complex (axoneme) and microtubulus sheath; 3) in the development of up to 4 bundles of outer fibrils in one spermatid. The increase of Fe content, lipid peroxidation and the onset of morphological alterations occurred already at a mild degree of Mg deficiency.Testis

ISSN:0303-4569    

DOI:10.1111/j.1439-0272.1996.tb02756.x

出版商:Blackwell Publishing Ltd    

年代:1996

数据来源: WILEY

摘要:

Book reviewed in this article:Enke, 1993. 3. bearb. Aufl., VIII, 206 Seiten, 5 Abbildungen, 2 Tabellen, kartoniert DM 68,‐. ISBN 3–432–25443–1.J. Baltzer, H. Mickan:Gynäkologie—ein kurzgefaßtes Lehrbuch. Thieme, 1993. 736 Seiten, 274 Abbildungen, 35 Tabellen, flex. TB DM 54,‐. ISBN 3–13–460605–4.M. Grillo:Experimentelle Untersuchungen zum Einfluß von Escherichia coli und Ureaplasma urealyticum auf die Fertilität.Thieme, 1994. 156 Seiten, 24 Abbildungen, DM 54,‐. ISBN 3–13–132901–7L. S. Neinstein:Issues in Reproductive Management.Thieme, 1994. X, 246 pages, 16 tables, cloth DM 70,‐. ISBN 3–13–119001–9.W. H. Weiske:Infertilität beim Mann.Thieme, 1994. VIII, 74 Seiten, 42 Abbildungen, 13 Tabellen, gebunden, DM 128,‐. ISBN 3–13–133 101–1C. Wernz:Sexualität als Krankheit.Ein medizinischer Diskurs zur Sexualität um 1800 (Beiträge zur Sexualforschung, Bd. 67). Enke, 1993, 328 Seiten, 4 Ab

ISSN:0303-4569    

DOI:10.1111/j.1439-0272.1996.tb02757.x

出版商:Blackwell Publishing Ltd    

年代:1996

数据来源: WILEY

ISSN:0303-4569    

DOI:10.1111/j.1439-0272.1996.tb02758.x

出版商:Blackwell Publishing Ltd    

年代:1996

数据来源: WILEY