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PubMed1995

Inhibition of bacterial growth in vitro by human ejaculate seminal plasma fraction 1 and its possible practical use in male infertility phenomenon.

PURPOSE: The in vitro bacteriostatic effect on bacteria by a fraction of human ejaculate (HuSePl-Fr1) was studied. METHOD: As bacterial indicators, Pseudomonas aeruginosa (Pa), total coliforms (Tc), fecal coliforms (Fc), Escherichia coli (Ec), and Staphylococcus aureus (Sa) were used. These bacterial strains are known as agents that cause urinary and sexually transmitted infections (STI). RESULTS: Preliminary results suggest that HuSePl-Fr1 has a significant in vitro bacteriostatic effect on Pa and Sa at 42 degrees C. In addition, Pa was inhibited at 37 degrees C after 48 hr incubation. The remaining bacterial indicators were not affected. CONCLUSION: The results indicate a possible protective role of HuSePl-Fr1 in treatment of infertile men due to genital infections.

Culture Media, Conditioned↗
PubMed1997

Sperm count in ejaculates and after sperm selection with discontinuous percoll gradient centrifugation technique, as a prognostic index of IVF outcome.

Success of IVF with low sperm count will depend on the retrieval of the maximum number of normal motile sperms, and this makes the selection of an appropriate technique critical. Various in vitro methods have been developed for selecting human sperm cells. Sperm selection using percoll gradient has been reported to yield upto 60% motile and morphologically normal spermatozoa from a normal semen sample. In this study, we have attempted to determine a possible relationship between sperm count in ejaculates before and after selection with percoll gradient centrifugation on one hand, and fertilization, cleavage and pregnancy rates in an IVF program on the other. With increased sperm count at the time of IVF treatment using selected sperm, we observed higher fertilization, cleavage and gestational rates. In conclusion, sperm concentration before and after selection with percoll may be considered a prognostic parameter for the determination of fertilization potential and pregnancy rates in an IVF program.

Cell Division↗
PubMed2001

Demonstration and cytochemical analysis of anionic sites on ejaculated boar spermatozoa: a scanning electron microscopy study using cationised colloidal gold.

The plasmalemma of spermatozoa bears negative charges as is the case for most mammalian cells. This has been concluded from the sperm cell's electrophoretic behaviour and from labelling experiments with various cationic probes followed by transmission electron microscopy of ultrathin sections. An overall view of the cell surface, however, is necessary in order to assess the distribution and density of the anionic sites adequately. We, therefore, used scanning electron microscopy in combination with cationised colloidal gold labelling to analyse the presence of anionic sites on ejaculated boar spermatozoa. Incubations were performed at pH 3.5, 2.5 and 1.0. Labelling was specific and bound gold particles were unequivocally identified using the backscattered electron signal. The chemical nature of the anionic sites involved was investigated by treating spermatozoa with pronase, phosphatase and neuraminidase as well as by methylation, acid hydrolysis and beta-elimination prior to cationised gold labelling. Our results suggest that besides phosphates, carboxyl groups are predominantly accountable for the binding of cationised colloidal gold. Presumptive macromolecules bearing these anionic sites are phospholipids and sialic acid residues. The combination of methods presented herewith should be of value in order to elucidate charge interactions which have been shown to play a role in cellular recognition events and adhesion.

Acid Phosphatase↗
PubMed1986

Disaturated phosphatidylcholine as a fetal lung maturity test: lack of interference by seminal ejaculate as a contaminant in vaginally pooled amniotic fluid samples.

Seminal ejaculates obtained from four men were added to amniotic fluid pools. Lecithin/sphingomyelin ratios in amniotic fluid were significantly lowered in the presence of small amounts of contaminating semen. In contrast, the determination of disaturated phosphatidylcholine was not affected by the presence of this contaminant.

Amniotic Fluid↗
PubMed1989

Relationship between circulating antisperm antibodies in women and autoantibodies on the ejaculated sperm of their partners.

The relationship between antibodies on the surface of ejaculated sperm and circulating antibodies in female partners was evaluated. Of 616 couples examined by the immunobead binding test, there was a 12.4% incidence of sperm-surface antibodies in men whose wives had antisperm antibodies in their sera, but only a 6.5% incidence in partners of women who lacked these antibodies (p less than 0.025). Sperm-bound immunoglobulin G and immunoglobulin A both occurred at a significantly higher frequency (p less than 0.05) in partners of women with serum antisperm antibodies. Increased incidence of both immunoglobulin G (p less than 0.01) and immunoglobulin M (p less than 0.005) circulating antisperm antibodies in females were observed when the male partners had antibody-bound sperm. Antibody-coated sperm may activate lymphocytes in the female partners after coitus, thus leading to the production of antisperm antibodies. This may be an additional mechanism that leads to female isoimmunity to sperm and infertility.

Antibodies↗
PubMed1988

Motility, heat, and lactate production in ejaculated bovine sperm.

Effects of various inhibitors on motility, heat, and lactate production of ejaculated bovine sperm were determined in the presence of antimycin A and rotenone. erythro-9-[3-(2-Hydroxynonyl)]adenine (EHNA) and polyvinylpyrrolidone (PVP-360) stopped motility and reduced heat or lactate production by 30-50%. Carbodiimides resulted in loss of motility and a reduction of metabolism by 60-75%. Quercetin treatment, which enhanced rather than inhibited motility, depressed heat and lactate production by 50-60%. Since mechanical immobilization reduced heat production by only 30%, the question arises as to what other cellular processes are major contributors to the energy budget. Inhibitors of ion flux had little-to-no effect on heat or lactate production, suggesting that neither mitochondrial nor Na+/K+ ATPases were major ATP-requiring processes. Calcium flux at the plasma membrane also was minimal and previous reports eliminated glycolytic substrate cycling as major consuming processes for ATP. Although quercetin inhibited lactate production in intact cells, no effect of quercetin on cell-free glycolysis and the ATPase activities of isolated dynein was detected. Quercetin did, however, inhibit ATPase activity of plasma membrane, suggesting that this unidentified ATPase may contribute to the formation of ADP and Pi required for lactate production by the intact cell. We propose (a) that the bioenergetic costs of motility are divided between regulatory events and dynein-microtubule interaction (dynein ATPase), (b) that some of the membrane-related processes may be "inefficient," and (c) that quercetin may render these steps more "efficient," in a manner analogous to its action on the Na+/K+ pump of Ehrlich ascites tumor cells.

Adenosine Triphosphatases↗
PubMed1986

Control of respiration and of motility in ejaculated bull spermatozoa.

The relations between motility and respiration were studied in ejaculated bull spermatozoa respiring with lactate. Motility was quantitatively evaluated by a turbidimetric procedure as percentage of cells moving per minute from the bottom of the cuvette into the light path. For selective inhibition of ATP-consuming reactions including motility or of mitochondrial respiration, vanadate or cyanide, respectively, were used. Both inhibitors were found to produce proportional changes in motility and respiration. The simultaneous changes in motility and respiration were linked to shifts in the cellular ATP/ADP ratio. Partial uncoupling of respiration in vanadate-inhibited cells gave similar relations between respiration and ATP/ADP ratios as stepwise inhibition of ATP-utilizing reactions by vanadate. Presuming saturation kinetics with respect to the ATP/ADP ratio, half maximum constants of 1.7 and 4.7 for the ATP/ADP ratio and maximum values of about 130% and 300% (in comparison to untreated cells) were estimated for motility and respiration, respectively. Respiration showed a much steeper dependence on the ATP/ADP ratio than motility resulting in an apparent cooperativity coefficient of 2.9. From these dependences on the ATP/ADP ratio, the shares in the control of ATP turnover in untreated cells were estimated. At sufficient supply with substrate, more than 80% of control were excreted by motility and other ATP-utilizing reactions, the rest by mitochondrial ATP production, i.e., the reactions of oxidative phosphorylation.

Adenosine Diphosphate↗
PubMed1990

Gossypol-induced inhibition of glucose uptake in human ejaculated spermatozoa may be mediated by lipid peroxidation.

Gossypol, a plant-derived polyphenolic compound known to exert contraceptive actions in men, inhibits 14C-glucose uptake in vitro in human ejaculated spermatozoa. Spermatozoal glucose uptake was found to increase monotonically up to 40 min, and then decreased by 60 min, possibly because of the saturation of the transport loci in the membrane. Gossypol at the concentrations of 5 and 10 microns caused a reduction of both the linear portion of the uptake and the fall afterwards. Gossypol similarly affects both the Na-dependent and -independent glucose uptake. The kinetic parameters of glucose uptake indicate that gossypol might be interfering with the transport/carrier protein as reduction in maximum uptake velocity (Vmax) was observed without any change in the affinity constant (Km). Similarly, gossypol also produced an increase in spermatozoal lipid peroxidation as evidenced by a steep rise in thiobarbituric acid reaction products in the human sperm cells. A significant decrease in total phospholipid level and the individual classes was noted after gossypol addition. Gossypol-induced inhibition of glucose uptake may be related to the generation of lipid peroxides and consequent membrane damage.

Glucose↗
PubMed1994

Localization and structural characterization of an oligosaccharide O-linked to bovine PDC-109. Quantitation of the glycoprotein in seminal plasma and on the surface of ejaculated and capacitated spermatozoa.

PDC-109 (13 kDa) is the most abundant component, and the major heparin-binding protein, of bovine (Bos taurus) seminal plasma. Here, we show that PDC-109 contains a single O-linked oligosaccharide (NeuNAc alpha(2-6)-Gal beta(1-3)-GalNAc-) attached to Thr11. Immunoquantitation of PDC-109 indicates that its concentration in seminal plasma is 15-20 mg/ml. Though PDC-109 is not present on epididymal sperm, ejaculated spermatozoa on average are coated with (9.5 +/- 0.3) x 10(6) molecules of PDC-109/cell. This value remained constant in swim-up sperm and decreased to (7.7 +/- 0.4) x 10(6)/spermatozoon after incubation for 24 h in capacitation medium at 39 degrees C. These data substantiate the hypothesis that PDC-109 may be one of the seminal plasma components that enhance the fertilizing capacity of bull spermatozoa upon interaction with heparin-like glycosaminoglycans present in the female genital tract.

Amino Acid Sequence↗
PubMed1977

Immunoglobulin A in split ejaculates of patients with prostatitis.

7S and 11S IgA (immunoglobulin A) levels in split ejaculates of 17 patients with chronic prostatis and 16 normal subjects were studied by the Hanson radial immunogel filtration method. The results suggest at least in certain cases of prostatitis increase of IgA level involves elevation of both types of IgA, and they are mainly of prostatic origin.

Body Fluids↗
PubMed1985

Scrotal dimensions and ejaculate characteristics of three breeds of sheep in tropical Nigeria.

Scrotal circumference and semen characteristics of three breeds of sheep (Udda, Balami and Yankasa) indigenous to Nigeria and Southern Guinea Savannah zones of Africa were compared. The age, body weight, scrotal circumference and spermiogram of the rams were studied by standard techniques. The mean age, body weight, and scrotal circumference of the three breeds were not comparable with significant interbreed, but were with significant intrabreed differences. The mean ejaculate concentration of sperm cells (x 10 /ml) were: Udda, 3.8 +/- 0.050, Balami, 4.1 +/- 0.32, Yankasa, 4.5 +/- 0.11. The mean morphological sperm cell abnormalities for the Udda, Balami and Yankasa were; 7.5 +/- 2.1%, 4.5 +/- 0.58% and 6.0 +/- 0.87%, respectively, with significant inter- and intrabreed differences. There were significant intrabreed differences in the other semen traits, i.e., percent of live cells, percent of motility, mean volume and mean concentration. In all the breeds of sheep studied, the scrotal circumference and spermiogram were comparable to, and within the range reported for the exotic breed of rams.

Journal Article↗
PubMed1988

Two cases of multiple abnormalities in testicular and ejaculated bull spermatozoa.

Multiple abnormalities were observed in testicular, epididynal and ejaculated spermatozoa from two bulls. These included acrosomal knobbing, incomplete nuclear condensation and coiled tails. The first two defects are considered to be characteristic of emmission of immature sperm, while the last may be a reflection of the effect of a changing osmotic environment on inherently susceptible cells.

Journal Article↗
PubMed1990

Freezability, enzyme leakage and fertility of buffalo spermatozoa in relation to the quality of semen ejaculates and extenders.

Forty-eight semen ejaculates from four Surti buffalo bulls were studied under split sample technique to establish the effects of initial semen quality and tris fructose yolk glycerol (TFYF), egg yolk citrate glycerol (EYCG) and lactose yolk glycerol (LYG) extenders on the freezability, fertility (based on 3412 AI) and extracellular release of spermatozoal enzymes pre and postfreezing. The overall mean activity of GOT, GPT, AKP, ACP and LDH enzymes in the postthaw seminal plasma increased significantly (P<0.01) above prefreeze levels, whereas sperm motility decreased. Freezability and the release of these enzymes were not influenced by the types of extenders used except for AKP release, which was significantly (P<0.01) lower in TFYG diluent. The pre and postfreeze sperm motility was significantly higher (P<0.01) and the leakage of GOT, AKP and ACP was lower in 36 semen samples with an initial motility above 70% than in the 12 samples in which initial motility was between 60 and 70%. The effects of interactions between motility groups, diluents and freezing periods were statistically nonsignificant for both freezability and leakage of all five enzymes. Fertility rate of frozen semen produced in TFYG diluent was significantly (P<0.05) higher (42.69%) than in the other diluents and was followed by that of EYCG (39.78%) and LYG (37.50%) with an overall mean of 40%. Sperm post-thaw motility had significantly (P<0.01) negative correlations with the release of enzymes: GOT (-0.948); GPT (-0.859); AKP (-0.673); ACP (-0.951) and LDH (-0.764). Fertility rates showed high negative correlations with the release of all five enzymes. Freezability was positively correlated with fertility (+0.405). Significant positive correlations were also observed for the release of GOT with that of GPT (+0.944); AKP (+0.574); ACP (+0.911) and LDH (+0.839): GPT with ACP (+0.795) and LDH (+0.870): and ACP with AKP (+0.725) and LDH (+0.577). These findings stressed the use of simple estimates of GOT and AKP leakage as markers for the assessment of freezability and fertility, and also the importance of initial good quality semen and the suitablity of extenders (TFYG) in the production of frozen buffalo bull semen for better fertility rates.

Journal Article↗
PubMed1991

New data on aberrant spermatozoa in the ejaculate of Sus domesticus.

This paper describes 16 new types of aberrant spermatozoa observed by scanning electron microscopy in the ejaculate of two healthy, sexually mature Landrace boars. The new anomalies observed were 1) spermatozoa with folded tail and abnormal head; 2) tailless spermatozoa with an abnormal connecting piece; 3) immature spermatozoa with two tails of the same length, fused and coiled; 4) spermatozoa with two tails of the same length, fused and coiled, and a small, rounded head; 5) spermatozoa with two fused tails and a wide head; 6) spermatozoa with three tails of the same length, fused and coiled; 7) immature spermatozoa with two heads and two fused tails; 8) spermatozoa with two heads, one at each tip of the tail; 9) spermatozoa with a short, folded tail and a triangular head; 10) spermatozoa with a short tail lacking the intermediate piece; 11) spermatozoa with a short tail, without the main piece and with a long intermediate piece; 12) spermatozoa with a short tail, without the main piece and with a rough head; 13) spermatozoa with small, rounded head; 14) spermatozoa with small, aberrant heads; 15) spermatozoa with small, bacillary heads; and 16) immature spermatozoa with tapering heads.

Journal Article↗
PubMed1991

Detection of white blood cell populations in the ejaculates of fertile men.

Using a now established, reliable technique for detecting white blood cells in human semen, we have examined the ejaculates of a fertile group of men. We have also highlighted the need for strict recruitment criteria to be established to enable accurate comparisons to be drawn between different patient groups and results from different laboratories.

Antibodies, Monoclonal↗
PubMed1987

Comparative studies of elemental composition on ejaculated fowl, bull, rat, dog and boar spermatozoa by electron probe X-ray microanalysis.

1. Comparative analyses of the concentrations of bulk and trace elements on the head, midpiece and tail regions of the ejaculated fowl, bull, rat, dog and boar spermatozoa were performed using X-ray microanalysis with scanning electron microscopy. 2. Although the pattern of distribution of elements on the surface of the three different subcellular regions was, in general, similar among all the species, there were substantial shifts in absolute concentrations. 3. Concentration of magnesium on the dog spermatozoa was significantly higher (about 2 times) than those of the other species. 4. Zinc, copper, iron and manganese concentrations were higher on fowl spermatozoa compared with those of the other species studied. 5. The Na-to-K ratios on the midpiece ranged from 1.46 (rat) to 2.26 (dog).

Animals↗
PubMed2005

[Detection of apoptotic markers in human ejaculated spermatozoa as new methods in human reproductive biology].

Ejaculated spermatozoa, particularly in infertile men, have been shown to display numerous features that are typical of apoptosis in somatic cells including Fas expression, ROS production, activation of caspases, DNA fragmentation, reduction in mitochondrial membrane potential, plasma membrane translocation of phosphatidylserine and permeability. In this review we summarize the biological significance and the potential role of these markers in the exploration of men infertility.

Apoptosis↗
PubMed2006

Calving sex ratio as related to the predicted Y-chromosome-bearing spermatozoa ratio in bull ejaculates.

The first objective was to correlate calving sex-ratio data from semen lots with the semen sex ratio obtained by two duplex polymerase chain reaction (PCR)/gel electrophoresis techniques. The two techniques involved different starting DNA amounts, PCR conditions, agarose gel concentrations, sample placement on the gels, lane size, number of lanes per gel, and duration of electrophoresis. The second objective was to sequence the duplex PCR products to verify their match to genes and chromosomes for which they were designed. Thirty-six ejaculates (lots) from eight Holstein sires were collected. Semen straws were distributed among dairies in three states. Ten straws per lot were used for the different PCR techniques. Sperm DNA was extracted and PCR analysis was done using one primer set to amplify a single copy section of the factor IX precursor (X-chromosome only) and another primer set to amplify a single copy section the sex determining region (Y-chromosome only). The glyceraldehyde phosphate dehydrogenase gene was amplified as an internal control. Standard curves were designed using PCR products in known ratios. Gel electrophoresis and image analysis were used to determine predicted %Y-chromosome-bearing spermatozoa (PredPtY). Sex (male=1, female=0) was reported on 526 calves and the ratio of the number of male to total calves (proportion of male calves (PMC)) was determined between sire and lot within sire. The PredPtY and PMC were significantly correlated (r=0.82, P<0.0002). No significant variance between sires was found in PredPtY or PMC, but lots within sires was a significant variance source for both. The two PCR technologies adequately determined semen sex ratio. The technology-by-lot-within-sire interaction was a significant variance source for PredPtY. Acrosomal integrity (after a 2-h) incubation, was correlated with both PMC and PredPtY; other semen quality characteristics had no significant correlations with PMC or PredPtY. Therefore, calf crop sex ratio skewness could be controlled by screening semen for PredPtY through the use of PCR.

Animals↗