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PubMed1998

Assessment of ejaculate quality and sperm characteristics in turkeys: sperm mobility phenotype is independent of time.

Given a pool of genetically superior male turkeys, the subsequent selection of toms as semen donors for artificial insemination should be based upon criteria that are predictive of the tom's fertility and fecundity over the course of a production cycle. Recently, sperm mobility phenotype has been shown to correlate highly with fertility. Therefore, the objectives of this study were to determine: 1) whether sperm mobility phenotypes of individual toms were independent of time, and 2) the extent to which traditional semen quality variables changed as a function of time during the study interval. Sperm mobility phenotype was determined by objectively measuring the ability of sperm to penetrate 2% Accudenz solution at body temperature. During the initial assessments of a flock (n = 94), sperm mobility indexes (SMI) were significantly higher for the High Mobility Phenotype toms (56.61 +/-1.03% SMI) compared to the Low Mobility Phenotype (30.46+/-1.27% SMI, P < or = 0.0001) toms. Over the 5 mo of this study, the High Mobility Phenotype toms consistently had higher (P < or = 0.05) SMI values than the Low Mobility Phenotype toms, with SMI values similar to those of the initial screen. Ejaculate volume, concentration, and plasma membrane integrity showed no significant differences between phenotypes (P > or = 0.05). Sperm viability remained significantly lower (P < or = 0.05) over the course of this study for the High Mobility Phenotype toms than for the Low Mobility Phenotype toms. Because sperm mobility phenotype remained consistent through time, the Sperm Mobility Test provides a potentially important tool for selecting semen donors in turkeys.

Animals↗
PubMed1987

Effects of fluoride and caffeine on the metabolism and motility of ejaculated bovine spermatozoa.

Treatment of washed, ejaculated bovine sperm with 30 mM sodium fluoride immobilized the cells in a characteristically rigid form. In cells metabolizing endogenous substrates, fluoride decreased respiration by about 60%, but did not inhibit the cells' ability to produce adenosine-5'-triphosphate (ATP) via oxidative phosphorylation and did not block access to endogenous substrates. Fluoride-immobilized sperm maintained maximal ATP titers for at least 60 min, but oligomycin treatment rapidly depleted ATP, indicating that ATP synthesis and metabolism was occurring in immobilized sperm. The putative phosphodiesterase inhibitor caffeine (2.5 mM) restored motility and increased respiration in fluoride-treated sperm, but 8-bromo-adenosine-3',5'-monophosphate (8-bromo-cAMP) did not, even though 8-bromo-cAMP stimulated respiration in control (untreated) sperm. Carboxyfluorescein analysis of the intracellular pH of untreated sperm indicated a normal pH of 6.3. Fluoride addition decreased the apparent intracellular pH slightly, but this effect was attributable to dilution. Caffeine did not change internal pH in untreated or fluoride-immobilized sperm. Fluoride did not appear to affect cAMP metabolism, but caffeine increased intracellular cAMP titers by about 35% in both untreated and fluoride-inhibited sperm. However, caffeine treatment did not mimic 8-bromo-cAMP, as analyzed by electrophoresis and autoradiography of sperm proteins labeled with 32P from endogenously generated [32P]ATP. Clearly, caffeine is not stimulating motility in fluoride-treated sperm by affecting the cyclic AMP system. Fluoride also inhibited motility in digitonin-permeabilized sperm by a mechanism that may have involved magnesium depletion, but caffeine had no stimulatory effect on either untreated or fluoride-immobilized, permeabilized sperm.

8-Bromo Cyclic Adenosine Monophosphate↗
PubMed1995

Presence of endogenous nicks in DNA of ejaculated human spermatozoa and its relationship to chromomycin A3 accessibility.

During spermiogenesis, mammalian chromatin undergoes replacement of nuclear histones by protamines, resulting in a DNA that is highly condensed in the mature sperm. We have previously demonstrated that a percentage of human spermatozoa exhibit 1) positivity to the guanine-cylosine-specific chromomycin A3 (CMA3) fluorochrome and 2) the presence of endogenous nicks in their DNA. In situ protamination of mature human sperm limits the percentage of sperm positive to CMA3 and exhibiting endogenous nicks. In this study, we report further investigations that aim to clarify the relationship existing between levels of CMA3 stainability and the presence of endogenous nicks in the DNA of mature human spermatozoa. Human spermatozoa from 25 different samples showed values of sensitivity to the CMA3 fluorochrome ranging from 13% to 75%. The same samples showed a percentage of sensitivity to endogenous nick translation ranging from 1% to 38%. A strong correlation (r = 0.86) was evident between these two parameters. Prior staining of sperm with the CMA3 fluorochrome drastically reduced sensitivity to nick translation. In contrast, previously nick-translated sperm stained with CMA3 showed very little difference from samples that had not been pretreated. The presence of nicked sperm in the ejaculate may indicate anomalies during spermiogenesis and be an indicator of male infertility.

Biotin↗
PubMed1999

No evidence for killer sperm or other selective interactions between human spermatozoa in ejaculates of different males in vitro.

This study examines one of the possible mechanisms of sperm competition, i.e. the kamikaze sperm hypothesis. This hypothesis states that sperm from different males interact to incapacitate each other in a variety of ways. We used ejaculates from human donors to compare mixes of semen in vitro from the same or different males. We measured the following parameters: (i) the degree of sperm aggregation, velocity and proportion of morphologically normal sperm after 1 and 3 h incubation in undiluted semen samples, (ii) the proportion of viable sperm plus the same parameters as in (i) in 'swim-up' sperm suspensions after 1 and 3 h incubation, (iii) the degree of self and non-self sperm aggregation using fluorescent dyes to distinguish the sperm of different males, and (iv) the extent of sperm capacitation and acrosome-reacted sperm in mixtures of sperm from the same and different males. We observed very few significant changes in sperm aggregation or performance in mixtures of sperm from different males compared with mixtures from the same male and none that were consistent with previously reported findings. The incapacitation of rival sperm therefore seems an unlikely mechanism of sperm competition in humans.

Acrosome Reaction↗
PubMed1980

Influence of seminal plasma on the chromatin stability of ejaculated human spermatozoa.

Nuclear chromatin decondensation (NCD) of human ejaculated spermatozoa exposed to sodium dodecyl sulphate (SDS) has been studied. A high proportion of NCD reacting spermatozoa was only found in semen samples with a relatively low activity of some prostatic factor(s) (i.e. zinc/fructose ratio below 0.18) in the seminal plasma. Exposure to SDS for one h was found sufficient to reveal the main proportion of spermatozoa undergoing NCD in such a solution. Addition of seminal plasma with an apparently normal composition to a sperm population with a high NCD reactivity restored the sperm SDS resistance to normal, i.e. blocked the NCD-response. Other results indicated that NCD reactivity was decreased or abolished upon prolonged storage of the spermatozoa in the seminal plasma. The various results indicated that some factor(s) in the seminal plasma can preserve the nuclear chromatin stability of human spermatozoa and that this factor most likely is of prostatic origin.

Humans↗
PubMed1980

A new optical technique for evaluating collective motility of ram and bull ejaculated spermatozoa.

A new, non-destructive, objective technique for measuring collective motility of highly-concentrated ram and bull semen is described. The principle is based on changes in the reflected light scattered by motile spermatozoa. These changes can be recorded as a continuous analog wave pattern (Reflectospermiogram-RSG) and are correlated to the intensity of the turbulent motility as evaluated subjectively with an ordinary light microscope. Ram spermatozoa have, after ejaculation, a typical motility pattern, i.e., high, stable activity for about 20 min, then a period during which the motility decreases at a constant rate, and finally, a period with a low but rather constant activity. The usefulness of the technique has been demonstrated in various types of experiments.

Animals↗
PubMed1982

ATP and ADP content of human ejaculated spermatozoa. I. Relationship with semen physical parameters in normal donors and oligozoospermic patients.

ATP and ADP were measured by a bioluminescent method in human ejaculated spermatozoa from 31 normal donors and oligozoospermic patients. ATP and ADP expressed for the sperm total number were significantly correlated (P less than 0.001). ATP snd ADP levels were found significantly correlated with the total motile spermatozoa and with the total viable cells (respectively P less than 0.001 and P less than 0.001 for ATP and P less than 0.001 and P less than 0.001 for ADP). ATP/ADP ratio was found significantly lower in subjects with sperm motility less than 50% (P = 0.024) and total viable cells less than 80% (P = 0.026).

Adenosine Diphosphate↗
PubMed1984

Sialoglycoproteins of ejaculated human spermatozoa and seminal plasma.

Ejaculated human sperm were found to possess three major sialoglycoproteins with molecular weights of 30 000, 14 000 and 12 000 and one minor species of 18 000. Liquefied seminal plasma from normal donors contain two major sialoglycoproteins with molecular weights of 17 000 and 15 000 and two minor species of 70 000 and 54 000. In contrast, the liquefied sperm-free semen of vasectomized men had two major species of sialoglycoproteins of 20 000 and 18 000 daltons plus two minor species of 54 000 and 40 000.

Chromatography, Gel↗
PubMedDate not supplied

Studies of the correlation of morphological and biochemical parameters in human ejaculate in various andrological diagnoses; 2nd report: biochemical parameters.

In this report the authors present the results about biochemical analyses in human ejaculate. The following parameters were determined ATP, ADP, pyruvate, lactate, alpha-ketoglutarate, fructose, myokinase, phosphoglucoseisomerase, phosphoglycerate-kinase, ATPase, acid phosphatase. It was found a relationship between age and fructose content, this was equally for pyruvate, whilst the converse was observed for lactate. No age dependend relationship was observed for the other substrates. For ATP and ADP distinctly higher values can be demonstrated in normozoospermia. A special study of the extent to which the encyme activities in sperm plasma are associated with the respective pH values measured shows that pH values above 7.2 correlate with distinctly lower activities for acid phosphatase, PGI and MK.

Adenosine Diphosphate↗
PubMedDate not supplied

Cephalexin concentration in the human ejaculate following oral and parenteral administration.

Cephalexin levels in the ejaculates of 31 infection-free volunteers have been determined following the administration of 1 g of antibiotic. In 16 subjects the cephalexin was administered by intramuscular injection and in 15 by mouth. In each case the volunteers were divided into groups of five each of which received the drug, 1, 2 or 4 hours before the collection of semen. Antibiotic concentrations were determined microbiologically on agar plates using Sarcina lutea ATCC 9341. For selection purposes healthy subjects were considered to be those showing a normal anamnesis and physical exploration, an absence of leucocytes demonstrable by a cytomorphological evaluation of the semen, semen citric acid and fructose levels within normal limits and a qualitatively and quantitatively similar saprophytic flora to that seen in a culture of urine taken immediately before the collection of semen. Maximum concentrations were obtained 2 hours after the administration of the antibiotic and the concentration following intramuscular injection (6.9 microgram/ml) was significantly greater than that following oral administration (4.7 microgram/ml).

Administration, Oral↗
PubMedDate not supplied

Effect of seminal plasma and calcium on the stability of the surface protein composition of ejaculated bull spermatozoa.

Ejaculated bull spermatozoa were incubated in different media to elucidate the effect of seminal plasma and calcium on the stability of sperm surface compositions. The spermatozoa were radioiodinated prior to or after the incubation and the labelled proteins from the incubation media and spermatozoa were analysed by SDS-polyacrylamide gel electrophoresis. Seminal plasma detached efficiently sperm surface components. Even with concentration of 2% considerable amounts of the 44K and 17K proteins were released into the medium. The 17K proteins of seminal plasma were also adsorbed by sperm surfaces during incubation. The release of the 27K protein seemed to be dependent on calcium.

Animals↗
PubMedDate not supplied

Adenosine 5'-triphosphate (ATP) activity in normal and pathological human semen. Relationship with round cells of the ejaculate.

The aim of the present paper is deleted to determine ATP activity in the human seminal cells by bioluminescence and to investigate the relationship between these values and the number of round cells either peroxidase positive or peroxidase negative. We studied 244 untreated men, divided into six groups. Our mean value of ATP activity in normal semen was 20.02 +/- 0.65 n moles per 10(8) spermatozoa. In asthenozoospermic or oligoasthenozoospermic patients with less than 1,000,000 round cells per ml, the concentration of ATP was significantly lower than normal (P less than 0.001) or (P less than 0.05). Semen with normal characteristics or samples from asthenozoospermic or oligoasthenozoospermic patients but with more than 1,000,000 round cells, per ml and with predominance of peroxidase negative cells, had an ATP concentration higher than normal subjects (P less than 0.001). We believe that the knowledge of the quantity and quality of round cells of the ejaculate is important to interpret the seminal values of ATP.

Adenosine Triphosphate↗
PubMedDate not supplied

Effectiveness mechanism of chlomipramine by neurophysiological tests in subjects with true premature ejaculation.

The effectiveness mechanism of the chlomipramine treatment was evaluated by sacral evoked response (SER) and dorsal nerve somatosensory cortical evoked potential (DN-SEP) testings in 15 patients with true premature ejaculation (TPE). We couldn't demonstrate any significant difference between the values of either latency times or amplitudes of the evoked responses determined just before and at the end of the treatment with chlomipramine in these patients. However, the sensory thresholds were 24.4 +/- 4.3 V in the pretreatment term and 30.2 +/- 7.3 V at the end of the treatment. This difference is statistically significant (P = 0.0031). Our results suggest that chlomipramine increases the sensory threshold for the stimuli in the genital area.

Adult↗
PubMedDate not supplied

Correlation between multi-headed spermatozoa in the ejaculate and histological findings in the testis.

Semen smears of 163 patients as well as bioptic material of 163 patients (93 cases of paraffin histology and 70 cases of cryostat sections) were grouped by their content of multi-headed spermatozoa or multi-nucleated spermatids, respectively. It could be demonstrated that findings in the ejaculate concerning multi-headed spermatozoa are in good agreement with those in the testis in respect of the presence of double- and multi-nucleated spermatids. Furthermore, it could be shown that the frequency of multi-headed spermatozoa is positively correlated to inflammatory processes either of immunologic origin or caused by micro-organisms.

Cell Nucleus↗
PubMedDate not supplied

Percoll density gradient centrifugation and consecutive flow cytometry do not identify leukocytes and leukocyte subtypes in ejaculate specimens.

This paper describes an attempt to establish a new combined method of leukocyte analysis in human ejaculate by Percoll density gradient centrifugation and consecutive flow cytometry. As a first step, leukocyte separation was performed by Percoll density gradient centrifugation with consecutive enrichment of leukocytes, especially granulocytes, in the 40%/60% and 60%/80% Percoll interfaces. Then these fractions were stained with specific monoclonal antibodies and analysed in a Facscan flow cytometer. Flow cytometric analysis did not demonstrate identifiable leukocyte populations, indicating a questionable cross-reaction with spermatozoal elements. Therefore, the combined technique of Percoll density gradient centrifugation and flow cytometric analysis were considered unsuitable for clinical leukocyte determination.

Antibodies, Monoclonal↗
PubMedDate not supplied

Differentiation of ejaculates showing reactive oxygen species production by spermatozoa or leukocytes.

Differences between subgroups and correlations between reactive oxygen species (ROS), sperm motility, concentration of leukocytes and viability in semen samples from 143 men were investigated. Patients with azoospermia or leukocytospermia were excluded from the study. Spermatozoa were separated by means of glass wool filtration. Reactive oxygen species were determined by means of luminol chemiluminescence before and after sperm separation; thereafter, normozoospermic and oligozoospermic patients were divided into three subgroups using the mean of all patients investigated (17462 count 10(-7) viable spermatozoa) as cut-off value as follows: G1--high reactive oxygen species production in native semen and after glass wool filtration; G2--high production of reactive oxygen species only in native semen; G3--low levels of reactive oxygen species before and after glass wool filtration. In general, reactive oxygen species were significantly higher in oligozoospermic samples than in normozoospermic samples. In men with normal sperm count, ROS production correlated significantly with the number of leukocytes in the ejaculate before glass wool filtration, but not thereafter. Glass wool filtration is useful to distinguish between sperm samples in which reactive oxygen species are generated by leukocytes and those in which reactive oxygen species are excessively generated by the spermatozoa themselves.

Humans↗
PubMed1986

Season and social environment influence the membrane integrity of ejaculated boar spermatozoa as assessed by ouabain sensitivity.

Eight adult Landrace boars were housed for 12 months in one of two social environments. Socially nonrestricted boars were penned near estrual females and socially restricted boars were penned behind solid walls to eliminate visual and physical contact with other pigs. All animals were subjected to natural changes in day length. The sensitivity of ejaculated spermatozoa to ouabain (in inhibitor of Na+-K+ ATPase) was determined on 4 consecutive weeks in November, March-April, and July-August. Semen was diluted in Tyrode's solution (pH 7.4) with and without 10(-3) M ouabain. Duplicate samples of control and ouabain-treated spermatozoa were incubated at 37 degrees C for 4 h, and percent motile sperm, motility type, and motility index (combination of percent and type) were determined at hourly intervals. Ouabain-induced decreases in most motility parameters varied with season (season X treatment, P less than 0.05). At hour 4, induced decreases in percent motile sperm were more pronounced in November and July-August than in March-April for socially nonrestricted boars. Decreases in motility type were greater (P less than 0.05) in November and July-August than in March-April for socially nonrestricted boars and were greater (P less than 0.01) in November than in July-August for restricted boars. In March-April motility type decreased (P less than 0.01) to a greater extent for socially restricted vs. nonrestricted boars. Similar season and social environment differences were observed for motility index values. Given the interrelationships between ouabain sensitivity, the functional integrity of sperm cells, and fertilizing capacity, season and social environment differences in ouabain-induced motility depression probably reflect qualitative changes in boar spermatozoa.

Animals↗