Sperm Immobilization Research Articles

The N-linked carbohydrate moiety of male reproductive tract CD52 (mrt-CD52) interferes with the complement system via binding to C1q

Antisperm antibody detected in infertile female patients’ sera has been shown to correlate with reduced fertility. The antibody showed strong complement-dependent cytotoxicity as determined by the sperm immobilization test (SIT). CD52 is a human glycosylphosphatidylinositol (GPI)-anchored antigen present in lymphocytes and male reproductive tracts (mrt), including mature sperm and seminal plasma. Recently, purified mrt-CD52 from human seminal plasma has been reported to interfere with the classical complement pathway, but not lectin binding or alternative pathways of the complement system. The purpose of this study is to determine which stage of the classical pathway mrt-CD52 regulates. mrt-CD52 was purified from human seminal plasma or intact sperm membrane. Immunoprecipitation assay was performed with the reaction of mrt-CD52, human complement and mAb H6-3C4. Immunoprecipitate was formed by the carbohydrate moiety of mrt-CD52, but not by the GPI-anchor peptide. The C1q molecule (29kDa) was detected in the immunoprecipitates by Western blotting analysis probed with anti C1q antibody, indicating that the carbohydrate moiety of mrt-CD52 binds to C1q. Also, the complement-dependent SIT revealed that purified CD52 inhibited sperm immobilization activity by antisperm antibody. These results suggest that mrt-CD52 protects sperm function from complement attack if antisperm antibody is generated in the female reproductive tracts.

Do men with prostate abnormalities (prostatitis/benign prostatic hyperplasia/prostate cancer) develop immunity to spermatozoa or seminal plasma?

Prostate is an immunocompetent and not an immunoprivileged organ. It has an active immunologic armamentarium. There are three major prostate abnormalities namely, prostatitis, benign prostatic hyperplasia (BPH) and prostate cancer. In all these abnormalities, infection/inflammation has been implicated. As infection/inflammation of the male genital tract can also be involved in induction of antisperm antibodies (ASA), this study was conducted to examine if these prostate abnormalities lead to the formation of ASA. Sera were obtained from normal healthy men (n = 20), men with chronic prostatitis (n = 20), men with BPH (n = 25), men with prostate cancer (n = 25) and immunoinfertile men (n = 10). The presence of antisperm antibodies against lithium diiodosalicylate (LIS)-solubilized human sperm extract (HSE), seminal plasma and synthetic peptides based upon sperm-specific antigens namely fertilization antigen (FA-1) and YLP(12), were analysed using the sperm immobilization technique (SIT), tray agglutination technique (TAT), enzyme-linked immunosorbent assay (ELISA) and indirect immunobead binding technique (IBT). All the sera from normal men and men with prostate abnormalities (chronic prostatitis/BPH/prostate cancer) were found to be negative in SIT and TAT. In ELISA, a few sera from men having prostate abnormalities (4-24%) showed a weak positive immunoreactivity (2-3 SD units) with some of the spermatozoa/seminal plasma antigens. Majority of the samples did not show any immunoreactivity (<2 SD units) in ELISA. Even the samples that showed a weak positive immunoreactivity in ELISA did not bind to live human sperm in IBT, indicating lack of sperm binding antibodies in these sera. In all these assays, the sera from immunoinfertile men were positive. Our findings indicate that chronic prostatitis, BPH and prostate cancer do not induce antibodies to spermatozoa, sperm-specific antigens and seminal plasma components. Although prostate is an immunologically competent organ, and its abnormalities cause a rise in circulating prostate-specific antigen (PSA), it appears that there is no concomitant induction of immunity to spermatozoa/seminal components including sperm-specific fertility-related antigens, thus not causing ASA-induced immunoinfertlity. This is the first study to our knowledge reporting the absence of ASA in men with BPH and prostate cancer.

Imprisonment-Associated Sperm Clumping and Male Infertility

This observational, retrospective study investigated infertility among Iraqi men who had been long-term prisoners of war. Routine seminal fluid analyses were undertaken in former prisoners with suspected infertility attending the Ramadi Infertility Clinic, Ramadi, Iraq; semen samples were also microscopically evaluated in these patients. In 16 out of 38 patients (42%), male infertility was found to be associated with the formation of unusually large clumps of sperm. These sperm clumps contained large numbers of sperm; clumping led to sperm immobilization and infertility. The causes of sperm clumping are unclear. It is possible that the sperm clumping observed in the present study might be a result of developmental defects, possibly related to stress or other undiscovered factors, although this requires further investigation.

Human Sperm Interaction withStaphylococcus aureus: A Molecular Approach

Sperm immobilization factor (SIF) causing 100% immobilization of spermatozoa isolated from Staphylococcus aureus when characterized using LC-MS (Liquid chromatography-mass spectrometry) showed that this 20 kDa protein had peptide sequence similarity with hsp-70 protein. It was found to completely (100%) inhibit Mg++ ATPase activity of spermatozoa at concentration of 100 μg mL−1. Sperm samples treated with SIF also showed reduction in calcium ionophore-induced acrosome reaction as compared to control samples (treated with calcium ionophore alone). Binding studies of FITC labelled SIF with spermatozoa using fluorescent microscopy showed binding of SIF to the surface of spermatozoa indicating the presence of SIF binding receptor. The receptor was extracted by 3M NaCl and purified by gel permeation chromatography. Characterization of the receptor by MALDI-TOF (Matrix-assisted laser desorption ionization-time of flight) indicated that the receptor shared sequence similarity with MHC class II antigen. A calorimetric study showed that the receptor moiety on spermatozoa was specific for the purified ligand as binding of the receptor to ligand was enthalpically (−11.9 kJ mole−1) as well as entropically (21.53 J mole−1 K−1) favored resulting in the Gibb's free energy of −18.57 kJ mole−1.

Molecular Mimicry Between Spermatozoa and Bacteria

Sperm immobilization factor isolated from Staphylococcus aureus immobilizes human spermatozoa as well as motile bacteria, showing that sperm immobilization factor receptor might be shared by bacteria and human spermatozoa. Thus, we sought to identify a common sperm immobilization factor binding receptor on spermatozoa and bacteria. Sperm immobilization factor was isolated from S. aureus. Sperm immobilization factor binding receptors were isolated from spermatozoa and bacteria. Antisperm antibodies in humans cross-reacted with bacteria antibodies. Thus, molecular similarities between determinants of spermatozoa and pathogenic microorganisms can be inferred. Sperm immobilization factor isolated from S. aureus immobilized spermatozoa (150 μg/ml) and motile bacteria, including Escherichia coli, Pseudomonas aeruginosa and Proteus mirabilis (500 μg/ml). This showed the presence of a common sperm immobilization factor binding conformation on spermatozoa and bacteria. Furthermore, receptors binding sperm immobilization factor were extracted and purified from spermatozoa, E. coli, P. aeruginosa and P. mirabilis. Molecular mimicry between spermatozoa and bacteria was confirmed by observing the blockage of spermatozoa immobilization by sperm immobilization factor in the presence of receptors isolated from spermatozoa, E. coli, P. aeruginosa or P. mirabilis. Also, a higher concentration of sperm immobilization factor (200 μg/ml) caused sperm death. Blocking the death of spermatozoa induced by sperm immobilization factor in the presence of these receptors provided further evidence for a common receptor. Results provide evidence for molecular similarity between bacteria and spermatozoa.

Sperm Immobilization Properties of Aqueous Ethanolic Extract of &lt;i&gt;Hymenocardia Acida&lt;/i&gt; Stem Bark

Hymenocardia acida (Large red-heart) is widely used in Nigeria and many African countries, and has many therapeutic benefits. This study evaluated the sperm immobilization property of aqueous ethanolic extract of Hymenocardia acida stem bark. It was found that the extract of Hymenocardia acida reduced sperm motility in concentration-dependent manner. At 10 % concentration, the spermatozoa immediately became immotile. The results showed that Hymenocardia acida stem bark extract possesses sperm immobilization property.

Editorial Comment to Receptor dependent immobilization of spermatozoa by sperm immobilization factor isolated from Escherichia coli: Proof of evidence

International Journal of UrologyVolume 18, Issue 8 p. 603-604 Editorial Comment to Receptor dependent immobilization of spermatozoa by sperm immobilization factor isolated from Escherichia coli: Proof of evidence Decio Armanini M.D., Decio Armanini M.D. Department of Medical and Surgical Sciences, Endocrinology, University of PaduaSearch for more papers by this authorFrancesco L Giorgino M.D., Francesco L Giorgino M.D. A.G.E.O., Padua, Italy. decio.armanini@unipd.itSearch for more papers by this authorCristina Fiore M.Sc., Cristina Fiore M.Sc. Department of Medical and Surgical Sciences, Endocrinology, University of PaduaSearch for more papers by this author Decio Armanini M.D., Decio Armanini M.D. Department of Medical and Surgical Sciences, Endocrinology, University of PaduaSearch for more papers by this authorFrancesco L Giorgino M.D., Francesco L Giorgino M.D. A.G.E.O., Padua, Italy. decio.armanini@unipd.itSearch for more papers by this authorCristina Fiore M.Sc., Cristina Fiore M.Sc. Department of Medical and Surgical Sciences, Endocrinology, University of PaduaSearch for more papers by this author First published: 22 June 2011 https://doi.org/10.1111/j.1442-2042.2011.02801.xCitations: 1Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat No abstract is available for this article.Citing Literature Volume18, Issue8August 2011Pages 603-604 RelatedInformation

Receptor dependent immobilization of spermatozoa by sperm immobilization factor isolated from Escherichia coli: Proof of evidence

To examine the receptor-ligand interaction between E. coli and spermatozoa resulting in sperm immobilization. Sperm immobilization factor (SIF) was isolated and purified from filtrate of E. coli suspension. Receptor on human spermatozoa for SIF was made to isolated and purified by salt extraction, gel permeation and ion exchange chromatography. Receptor dependent immobilization of spermatozoa by SIF was confirmed by competitive inhibition by addition of the purified receptor and binding to sperm receptor by Fluorescin Isothiocynate (FITC) labelled SIF using fluorescence microscopy. Heat labile, non-dialyzable, sperm immobilization factor of ∼56 kDa was isolated and purified from E. coli. Using SIF as a tool, receptor of 113 kDa could be isolated and purified from human spermatozoa. Addition of purified receptor completely inhibited sperm immobilization and death induced by SIF suggesting receptor-dependent immobilization of spermatozoa. Further incubation of sperm with FITC labelled SIF resulted in staining of whole sperm. The study provides evidence of receptor-ligand interaction between E. coli and spermatozoa.

Robotic ICSI (Intracytoplasmic Sperm Injection)

This paper is the first report of robotic intracytoplasmic sperm injection (ICSI). ICSI is a clinical procedure performed worldwide in fertility clinics, requiring pick-up of a single sperm and insertion of it into an oocyte (i.e., egg cell). Since its invention 20 years ago, ICSI has been conducted manually by a handful of highly skilled embryologists; however, success rates vary significantly among clinics due to poor reproducibility and inconsistency across operators. We leverage our work in robotic cell injection to realize robotic ICSI and aim ultimately, to standardize how clinical ICSI is performed. This paper presents some of the technical aspects of our robotic ICSI system, including a cell holding device, motion control, and computer vision algorithms. The system performs visual tracking of single sperm, robotic immobilization of sperm, aspiration of sperm with picoliter volume, and insertion of sperm into an oocyte with a high degree of reproducibility. The system requires minimal human involvement (requiring only a few computer mouse clicks), and is human operator skill independent. Using the hamster oocyte-human sperm model in preliminary trials, the robotic system demonstrated a high success rate of 90.0% and survival rate of 90.7% (n=120).

Spermicidal action of a protein isolated from ethanolic root extracts of Achyranthes aspera: An in vitro study

A previous study conducted in our department, showed that 50% ethanolic extract of the roots of Achyranthes aspera possess spermatotoxic effects. Preliminary studies also revealed that the active principle may be a protein. In this study a 58 kDa Achyranthes protein (Ap) was isolated from Achyranthes aspera using standard protocols and their effects on the rat sperm was studied in vitro in comparison with nonoxynol-9 (N-9). The sperm immobilization studies showed that about 150 μg of Ap was able to immobilize sperms completely within seconds at a lower concentration than N-9 (250 μg). The sperm revival test revealed that the spermicidal effect was irreversible. There was also a significant reduction in sperm viability and hypo-osmotic swelling in the Ap-treated and N-9 treated groups in comparison to the control. In the Ap and N-9 treated groups the number of acrosome reacted cells were found to be high and it also caused agglutination of the sperms indicating the loss of intactness of the plasma membrane which was further supported by the significant reduction in the activity of membrane bound 5' nucleotidase and acrosin enzyme. Hence this study showed that the protein isolated from the roots of Achyranthes aspera possess spermicidal activity in vitro and can act as a spermicide similar to that of nonoxynol 9. Ap also possessed spermicidal activity against human sperms in vitro.

Incidence of Sperm-immobilizing Antibodies in Infertile Women with Past Chlamydia trachomatis Infection

Among the risk factors for antisperm antibody production, inflammatory diseases of the genital tract are believed to play an important role. Chlamydia trachomatis infection is one of the most common sexually transmitted diseases. There are some reports suggesting that human sperm have antigens that cross-react immunologically with certain microbial antigens, such as C. trachomatis. However, this is still controversial. We performed a retrospective study to investigate the correlation between anti-chlamydial antibodies and sperm-immobilizing antibodies in infertile women. Between January 2007 and March 2009, the presence of sperm-immobilizing antibodies was examined by the sperm immobilization test using sera from 273 infertile women. Anti-chlamydial antibodies (IgG and IgA) were examined to prove past C. trachomatis infection by ELISA using the same sera from infertile women. The overall incidence of sperm-immobilizing antibodies was 2.9% (8/273) in infertile women. The incidences of sperm-immobilizing antibodies were 6.4% (5/78) in cases with past C. trachomatis infection and 1.5% (3/195) in cases without past C. trachomatis infection. There was a significant difference between the two groups (P = 0.031). A significantly higher incidence of sperm-immobilizing antibodies was noted in infertile women with past C. trachomatis infection compared with that of those without past C. trachomatis infection. This is the first demonstration that C. trachomatis infection could play a role in the production of sperm-immobilizing antibodies in infertile women.

Automated Sperm Immobilization for Intracytoplasmic Sperm Injection

Sperm immobilization is a requisite step in intracytoplasmic sperm injection (ICSI). Conventionally, sperm immobilization is performed manually, which entails long training hours and stringent skills. Manual sperm immobilization also has the limitation of low success rates and poor reproducibility due to human fatigue and skill variations across operators. This paper presents a system for fully automated sperm immobilization to eliminate limitations in manual operation. Integrating computer vision and motion control algorithms, the automated system is able to visually track a sperm and control a micropipette to immobilize the sperm. A robust sperm tail tracking algorithm is developed to locate the optimal position on the sperm tail for sperm immobilization. The system demonstrates: 1) an average sperm tail tracking error of 0.95 μm; 2) a sperm tail visual tracking success rate of 96%; 3) a sperm immobilization success rate of 88.2% (based on 1000 trials); and 4) a speed of 6-7 s per successful immobilization.

Mechanism for sperm immobilization activity of extract from Platycodon grandiflorum in vitro

To explore the mechanism of spermicidal effect of crude extract and platycodin-D from Platycodon grandiflorum (PG) root in vitro. Between February 2006 and December 2009, 38 fertile and healthy adult males were selected as donors. PG root was extracted and platycodin-D purified. Grouping was as follows: crude extract from PG root, platycodin-D, nonoxynol-9 (N-9, as a reference standard) and semen-added physiological saline (as control). Spermicidal experiments were carried out in vitro (Sander-Cramer test). The hypo-osmotic swelling (HOS) test and modified Eosin-Giemsa (EG) staining were used to detect the integrity of sperm membrane. Four types of sperm morphology were divided through HOS-EG test: Type A: spermatozoa with swelling in tails and head white staining HOS(+)-EG(-) (membrane intact); Type B: spermatozoa with no swelling in tails (membrane-damaged) and head white staining HOS(-)-EG(-); Type C: spermatozoa with tail swelling and head red HOS(+)-EG(+); Type D: spermatozoa with no swelling in tails and head red HOS(-)-EG(+). Sperm chromatin dispersion (SCD) test was performed to determine the integrity of sperm DNA. The crude extract from PG root could immobilize and kill 100% spermatozoa within 20 s in vitro at the concentrations of 50.0 g/L and 20.0 g/L (v:v = 1:1 in semen). When the semen sample was exposed to the concentrations of 2.0 g/L and 1.0 g/L of platycodin-D, all spermatozoa were immobilized within 20 s. In the control group, the mean percentage of Types A, B, C and D was (69.0 ± 8.3)%, (3.4 ± 0.5)%, (10.2 ± 1.7)% and (17.4 ± 2.1)% respectively. In the groups of platycodin-D and N-9 solution, the rate of Types A and B was 0. The rate of Types C [(65.3 ± 3.8)%] and D [(34.7 ± 7.1)%] significantly increased versus control in the platycodin-D group (P < 0.01). Sperm DNA fragmentation had no change upon an exposure to the extract from PG root, platycodin-D and N-9 solution. And the sperm revival test showed none of the spermatozoa recovered their motility. The extract and platycodin-D from PG root have a quick sperm-killing effect in a short time in vitro by disrupting the integrity of sperm membrane (main head).

Automated sperm immobilization for human ICSI

OBJECTIVE: To develop a robotic system capable of performing automated sperm immobilization with a high success rate for clinical ICSI use.DESIGN: The robotic system consists of: (i) an XY stage that is visually servoed to move the sperm of interest to the center of the field of view; (ii) a novel sperm tail visual tracking algorithm designed to locate a point on the sperm tail for a micropipette to tap; and (iii) optimal velocity and motion planning of the micropipette for tapping the sperm tail to achieve sperm immobilization.MATERIALS AND METHODS: Trials on 1,000 sperm from patients were performed with the automated system. The sperm were in a viscous culture medium (SpermCatch, Nidacon).Table 1Automated sperm immobilization results.Sperm speed v (microns/sec)Immobilized/totalabcde% Immobilized40 < v <= 5566/811222881.525 < v <= 40301/345311417987.215 < v <= 25261/3151151817382.95 < v <= 15189/1910011098.50 < v <= 565/682100095.6total882/100072925372088.2 Open table in a new tab CONCLUSION: The automated robotic system is capable of conducting human sperm immobilization with a high success rate. It is a critical procedure for automated human ICSI. OBJECTIVE: To develop a robotic system capable of performing automated sperm immobilization with a high success rate for clinical ICSI use. DESIGN: The robotic system consists of: (i) an XY stage that is visually servoed to move the sperm of interest to the center of the field of view; (ii) a novel sperm tail visual tracking algorithm designed to locate a point on the sperm tail for a micropipette to tap; and (iii) optimal velocity and motion planning of the micropipette for tapping the sperm tail to achieve sperm immobilization. MATERIALS AND METHODS: Trials on 1,000 sperm from patients were performed with the automated system. The sperm were in a viscous culture medium (SpermCatch, Nidacon). CONCLUSION: The automated robotic system is capable of conducting human sperm immobilization with a high success rate. It is a critical procedure for automated human ICSI.

Temperature-dependent regulation of sperm motility of Ijima's copper pheasants ( Syrmaticus soemmerringii ijimae), one of ‘near threatened’ species

In order to conserve the copper pheasants, one of the Japanese ‘near threatened’ species, the knowledge on the sperm characteristics is the inevitable issue. Therefore, temperature-dependent regulation of copper pheasant sperm motility was investigated in comparison with that of domestic fowl spermatozoa. Motility of intact spermatozoa from both species was markedly affected by temperature. During incubation at 30 °C, copper pheasant spermatozoa showed around 60–70% motility, but became almost immotile when the temperature was raised to 40 °C. Then, when the temperature of the sperm suspension was subsequently cooled to 30 °C, the spermatozoa regained their motility. The motility of domestic fowl spermatozoa showed a similar pattern. Temperature also affected the motility of both demembranated copper pheasant and domestic fowl spermatozoa in the same way. The motility of intact copper pheasant and domestic fowl spermatozoa at 30 °C was unaffected following the addition of 2 mM CaCl 2, 100 nM calyculin A, an inhibitor of protein phosphatase-type 1 (PP1), or 4 mM diB-cAMP, respectively, compared with those with no effectors. However, the presence of 10 μM ML-7, a selective inhibitor of myosin light chain kinase (MLCK), inhibited motility of spermatozoa from both species. At 40 °C, the presence of CaCl 2 or calyculin A restored the motility of spermatozoa from both species, but the addition of diB-cAMP or ML-7 could not prevent the immobilization of spermatozoa. At 30 °C in the presence of ATP, the motility of demembranated copper pheasant spermatozoa was over 60% but was inhibited following the addition of 10 μM ML-7; a similar pattern was found with demembranated domestic fowl sperm motility. The motility of demembranated spermatozoa from both species was inhibited following the addition of 2 mM EGTA to the reactivation medium at 30 °C, but restored by the subsequent addition of 4 mM CaCl 2. These results suggest that copper pheasant sperm motility might be regulated by similar mechanisms to that of domestic fowl spermatozoa: i.e., the balance of Ca 2+/MLCK or an MLCK-like protein-dependent phosphorylation and PP1-dependent dephosphorylation. The similarity in physiological regulation of spermatozoa from both species shows that extensive technology developed for artificial breeding of the domestic fowl might be applicable to captive breeding of copper pheasants.

Mechanism of Sperm Immobilization byEscherichia coli

Aim. To explore the influence of Escherichia coli on the motility of human spermatozoa and its possible mechanism. Methods. Highly motile preparations of spermatozoa from normozoospermic patients were coincubated with Escherichia coli for 4 hours. At 1, 2 and 4 hours of incubation, sperm motility was determined. The factor responsible for sperm immobilization without agglutination was isolated and purified from filtrates. Results. This report confirms the immobilization of spermatozoa by E. coli and demonstrates sperm immobilization factor (SIF) excreted by E. coli. Further this factor was purified by ammonium sulfate precipitation, gel permeation chromatography, and ion-exchange chromatography. Purified SIF (56 kDa) caused instant immobilization without agglutination of human spermatozoa at 800 μg/mL and death at 2.1 mg/mL. Spermatozoa incubated with SIF revealed multiple and profound alterations involving all superficial structures of spermatozoa as observed by scanning electron microscopy. Conclusion. In conclusion, these results have shown immobilization of spermatozoa by E. coli and demonstrate a factor (SIF) produced and secreted by E. coli which causes variable structural damage as probable morphological correlates of immobilization.

Comparative study on the spermicidal activity of organic solvent fractions from hydroethanolic extracts of Achyranthes aspera and Stephania hernandifolia in human and rat sperm

Background This study was conducted to determine the most effective fraction of the hydroethanolic (water:ethanol, 1:1) extracts of Stephania hernandifolia leaves and Achyranthes aspera roots (in a composite manner at a ratio of 1:3, respectively) that will provide maximum spermicidal activity in human and rat spermatozoa out of five different ratios (1:1, 1:3, 1:7, 3:1 and 7:1) that have been studied in pilot experiments. Study Design n-Hexane, chloroform and ethyl acetate fractions of the hydroethanolic (1:1) extracts of S. hernandifolia and A. aspera were mixed at 1:3. Different concentrations were tested for sperm immobilization, sperm viability, acrosome status, 5′-nucleotidase activity and nuclear chromatin decondensation using human and rat spermatozoa for the selection of the most effective concentration. Results Out of three fractions of the hydroethanolic (1:1) extracts of the said plants, the n-hexane fraction was most effective, and the chloroform fraction exhibited minimum activity for this purpose. At a concentration of 0.1 g/mL hexane fraction, all sperm of the human sample were immobilized immediately (within 20 s). In case of the rat sample, all epididymal spermatozoa were immobilized immediately (within 20 s) by treatment with hexane fraction at a concentration of 0.004 g/mL. All human sperm were found to be nonviable within 20 min. The activity of acrosome enzymes was reduced, and significant release of 5′-nucleotidase (a plasma membrane marker) into the surrounding medium was noted after treatment with 0.1 g/mL hexane fraction, indicating that the hexane fraction affected the cytoarchitecture of the sperm plasma membrane. The maximum number of human sperm failed to decondense when treated with 0.1 g/mL hexane fraction, and sperm motility was also irreversible. The hexane fraction was tested in rats as vaginal contraceptive and showed 100% efficacy, indicating its potential for development as vaginal contraceptive. Conclusion The findings indicate that, among the different fractions, the hexane fraction of the hydroethanolic extracts of the two plants produced the most effective spermicidal activity and can be considered as vaginal contraceptive.

Isolation of a spermatozoal immobilization factor from Staphylococcus aureus filtrates

Staphylococcus aureus isolated from the cervix of an infertile woman was found to cause complete immobilization of human spermatozoa in vitro. Only the cell culture and cell-free supernatant showed immobilization activity, indicating that the sperm immobilization factor might be released extracellularly by the organism because no activity was observed with the washed cells. Heat treatment of the supernatant at 60 degrees C for 10 min waived its immobilizing activity, indicating that the active component may be a protein. The bioactive molecule from the supernatant was purified to homogeneity by ammonium sulfate precipitation, gel permeation chromatography, and ion exchange chromatography. Sperm immobilization factor (SIF) was found to be an approximately 20 kDa protein. SIF at a concentration of 10 microg/mL was required to cause 100% immobilization of human spermatozoa after 30 min of incubation at 37 degrees C, whereas a concentration of 150 microg/mL caused immediate immobilization, and a concentration of 200 microg/mL resulted in instant loss of viability of human spermatozoa, observed by eosin-nigrosin staining. Scanning electron microscopy showed that the treatment of human spermatozoa with SIF caused multiple defects in the head, midpiece, neck, and tail region of human spermatozoa.

Evaluation of the immunological cause of male infertility.

The presence of antisperm antibodies was considered as one of the main causes of male infertility. This study was conducted on 776 semen samples of the male partners of infertile couples in order to evaluate the immunological factor. The detection of antisperm antibodies was performed by means of the direct Mixed Antiglobulin Reaction (MAR) test. A positive test was found in 35 samples. Hence, the prevalence of immunological infertility in this study is 4.5% among the male partners of infertile couples. Further explorations were performed on men with positive results, in order to detect the presence of antibodies in the sera by means of the indirect MAR tests, and sperm immobilization tests for quantitating the cytotoxic activity of the antibodies. The indirect MAR tests showed that 97% of the antibodies were present in sera, whereas 51% of the samples were cytotoxic. Routine semen analysis indicated that around 60% of the semen samples with antisperm antibodies were good. Semen culture showed that positive results were detected in 40% of samples. Clinical evaluations revealed that the presence of antisperm antibodies may have been the cause of long-standing infertility, on average 5 years, and were not all primary. Evaluations of all clinical and laboratory findings indicated that unilateral testis obstruction and male accessory gland infection were the main pathologies found.

Die Beeinflussung der Penetrationsrate humaner Spermatozoen im zona pellucida-freien Hamster-Oozyten-System durch monoklonale Antikörpern gegen humane Spermatozoen

The penetration rate of human spermatozoa in zona pellucida-free hamster oocytes was reduced by 4 monoclonal anti-sperm-antibodies (A-24, B-20, III-3, and VII-5) out of 6 tested. The other 2 (VI-1 and VI-16) left the penetration rates virtually unchanged. The extent of penetration rate reduction depended on the concentrations of antibodies A-24, B-20, III-3, and VII-5; if undiluted, they produced a significant reduction of penetration rates, the largest average reduction being almost 50% with undiluted antibody A-24. Decreasing concentrations of the monoclonal anti-sperm-antibodies resulted in significantly smaller reductions of sperm penetration rates. The reason for the reduced penetration rates observed may be a blockage of receptors on the sperm surface by the named monoclonal anti-sperm-antibodies. This can inhibit enzyme reactions of the spermatozoa, or interfere with the interaction between oocyte and spermatozoa membranes. It is noted that only the antibody III-3 caused agglutination of human spermatozoa in the micro-sperm-agglutination and immobilization tests. Therefore, the agglutination or immobilization of spermatozoa may not be held responsible for the reduced penetration rates after addition of monoclonar antibodies, as opposed to the findings with polyclonal antibodies (Steinberg - 1985&).