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Statistical andrology: standard semen analysis and computer-assisted sperm motility analysis.

Results are reported of a statistical evaluation of the variables obtained by standard microscopic semen analysis and by computer-assisted sperm motility analysis. Descriptive statistical analysis and cluster analysis were carried out on both sets of data. A contingency table was created to compare the classifications obtained by the two clustering procedures. A canonical correlation analysis was performed to evaluate the classification power of the variables obtained by the two semen analyses. The results show that the two techniques of semen analysis are complementary and not substitutive and that the correlated variables "percentage forward motility" and "sperm velocity" are the most discriminating.

Humans↗

Characterization of epithelial cell culture from human hydrosalpinges and effects of its conditioned medium on embryo development and sperm motility.

BACKGROUND: Recent studies have reported the negative impact of hydrosalpinx on IVF outcome. Toxic effects of hydrosalpinx fluid (HF) have been the main reason for the recommendation of functional surgery, salpingectomy, prior to IVF. The present study characterized hydrosalpinx epithelial cell culture and examined the effects of its conditioned medium (CM) on sperm motility, acrosome reaction and embryo development. METHODS: Normal Fallopian tubes (n = 6) and hydrosalpinges (n = 9) were used to prepare epithelial cell culture and CM. Epithelial cell characterization was confirmed using electron microscopy. Sperm motility and acrosome reaction were determined using computer-aided sperm analysis and acrobead assay respectively and embryo development by mouse embryo development assay. RESULTS: The percentage of human motile sperm incubated in hydrosalpinx CM was significantly different from those in normal Fallopian tube (NFT) CM and modified human tubal fluid medium (hTF) (control) (P < 0.05 at 3 h and P < 0.001 at 5 and 24 h), with alteration in movement characteristic, linearity, 24 h after incubation in hydrosalpinx CM (P < 0.05). However, other sperm movement characteristics remained unchanged. Reduced acrosome reaction and poor mouse embryo development were also observed in hydrosalpinx CM but not in NFT CM and hTF. CONCLUSIONS: The results suggest that hydrosalpinx epithelial cells may be producing a fluid milieu hostile to sperm and early embryo development. The established epithelial cell culture system may provide a model to further investigate the mechanisms underlying the toxic effects of HF on embryo development and the adverse effects on IVF outcomes.

Acrosome Reaction↗

[The relation between sperm motility and the content of unsaturated fatty acids as well as the osmotic behavior of human spermatozoa].

The relations between sperm motility and the content of lipids and fatty acids in sperm cells were investigated. In addition the osmotic volume changes of sperm cells were studied, by determination of progressive sperm motility, measurement of relative concentration of lipid-peroxy radicals in seminal plasma using chemoluminescence technique, gas-chromatographic determination of fatty acid pattern in sperm cells and in seminal plasma and by electronic measurement of the sperm cell volume. The impulse installment of chemoluminescence increased during the time of storage of the ejaculate. In contrast, this phenomenon was not observed in pure seminal plasma prepared by centrifugation immediately after liquefaction. The pattern of phospholipids and free fatty acids changed during storage of the ejaculate both in sperm cells and in seminal plasma. Sperm cells showed a significant increase of volume under hypotonic stress. The increase of volume is more distinct in spermatozoa with high progressive motility as in spermatozoa with low progressive motility. The determination of initial and late progressive motility enables the differentiation of two groups of sperm populations: one group with a medium initial motility and a normal decrease of motility during the following time and another group with excellent initial motility and an extreme decrease of motility following. The second group is characterized by an especially distinct increase of volume under hypotonic conditions.

Fatty Acids, Unsaturated↗

The effects of coital lubricants on sperm motility in vitro.

Infertility affects approximately 15% of couples, and in about one-third the primary cause is a male factor. Patients undergoing infertility investigations frequently experience sexual dysfunction, which often is due to inadequate vaginal lubrication. This can lead to increased use of coital lubricants. The effects of such lubricants on sperm motility have not been widely studied, although sperm motility is one of the best prognostic indicators of fertilization. Using a prospective longitudinal control-based study, we analysed the effect of adding four lubricants: KY jelly, baby oil, olive oil and saliva on sperm motion in 16 samples from patients undergoing infertility investigations. Sperm samples were prepared by density gradient centrifugation prior to mixing with lubricants. Motility parameters were determined using computer-assisted semen analysis after 5, 15 and 30 min. All lubricants except baby oil significantly decreased percentage progressive motility, progressive velocity, curvilinear velocity and lateral head displacement at 12.5% concentration. At a lower concentration of 6.25%, both olive oil and saliva still significantly reduced progressive motility parameters, while KY jelly diminished head movement parameters. Hence, even at these very low concentrations, coital lubricants impair sperm motility and thus may adversely affect fertility.

Cellulose↗

Stimulation of bovine sperm motility and respiration by the triazine dye cibacron blue F3GA.

Bovine sperm motility and respiration were stimulated by the triazine dye Cibacron Blue F3GA (CB), which may operate as a nucleotide mimic. CB stimulation of respiration was half-maximal at about 35 microM and respiration reached maximal levels about 1.5 minutes after CB addition. Respiratory stimulation was preceded by a transient increase in cytosolic cAMP. Sperm cAMP titers were elevated from 5 to 10 pmoles/10(8) cells within 30 seconds of CB addition, but rapidly dropped to a stable level of about 7.5 pmoles/10(8) cells. CB was a potent inhibitor of sperm membrane adenylyl cyclase and inhibited respiration in permeabilized cells. Taken together, the data indicated that CB stimulation was not manifested via the cytosol. In addition, a nonpermeant blue dextran preparation synthesized with CB also stimulated sperm respiration and motility. CB inhibited sperm membrane phosphodiesterase activity, suggesting that the transient pulse of cAMP resulted from CB interaction with this enzyme in the sperm membrane.

Adenylyl Cyclase Inhibitors↗

Use of computer-assisted sperm motility assessment and multivariate pattern analysis to characterize ejaculate quality in Mohor gazelles (Gazella dama mhorr): effects of body weight, electroejaculation technique and short-term semen storage.

Subjective and objective semen assessments were performed on 18 male Mohor gazelles (Gazella dama mhorr). Sperm motility assessments combined with sperm plasma membrane and acrosomal integrity evaluations were undertaken as part of a captive breeding programme. The primary objective was to test methodology for short-term preservation of gazelle semen for artificial insemination (storage in N-[Tris(hydroxymethyl)methyl]-2-aminoethanesulphonic acid-Tris diluent (TEST) for up to 96 h at 17 degrees C). However, the secondary objective was to investigate phenotypic and genotypic influences on semen quality within this small population, which was established in 1971 with only 12 genetic founders. Sperm motility was measured by computer-assisted semen assessment and the data were analysed using a pattern analysis technique to detect and quantify naturally occurring sperm subpopulations within the semen samples. Four sperm subpopulations distinguishable by their motion characteristics were detected. The relative frequencies of two subpopulations (population 2: highly motile, non-linear; and population 4: poorly motile, non-linear) in fresh semen were correlated with the maximum voltage used during electroejaculation. The frequency of subpopulation 2 was negatively correlated with maximum voltage (r = -0.875, P < 0.0001) and the frequency of subpopulation 4 was positively correlated (r = 0.727, P < 0.005). The frequencies of all subpopulations varied significantly among the animals sampled (chi-squared = 2577.6, degrees of freedom = 54, P < 0.0001) and subpopulation 4 was also correlated with body weight (r = -0.59, P < 0.005). Semen stored at 17 degrees C retained motility, plasma membrane and acrosomal integrity for 48 h, but these measures decreased thereafter. The frequency of a sperm subpopulation showing uncoordinated but active motility increased significantly over the first 48 h and then decreased.

Animals↗

[Changes of tumor necrosis factor alpha and interleukin-6 levels in peritoneal fluid of patients with endometriosis and their effects on human sperm motility and mouse embryo development].

OBJECTIVE: To study the changes of tumor necrosis factor alpha (TNF-alpha) and interleukin-6 (IL-6) levels in peritoneal fluid of patients with endometriosis and their effects on human sperm motility and mouse embryo development. METHODS: Levels of TNF-alpha and IL-6 in peritoneal fluid and cultured peritoneal macrophages supernatant were detected by enzyme-linked immunoassay (ELISA) in 14 infertile patients (EMT group) with endometriosis proven by laparoscopy and 11 infertile women with normal pelvic organs (control group). The effects of peritoneal fluids obtained from the 2 groups on human sperm motility and development of 2-cell mouse embryo were also studied in vitro. RESULTS: In EMT group the numbers of peritoneal macrophages were higher than those in the control group. Levels of TNF-alpha [(0.33 +/- 0.15), (7.19 +/- 0.60) micrograms/L] and IL-6 [(0.49 +/- 0.68), (1.51 +/- 0.08) micrograms/L] in peritoneal fluid and peritoneal macrophage supernatant were also elevated significantly as compared with those in the controls (P < 0.01). Meanwhile, after four-hour incubation with peritoneal fluids, the percentage of straight-line moving sperm and total sperm motility rate were decreased significantly in EMT group(P < 0.01). Appearance rate of 16-cell mouse embryo was 32.5% in EMT group versus 47.6% in control group after 48 hour coculture with peritoneal fluids(P < 0.01). CONCLUSION: It is likely that elevation of peritoneal fluids cytokines levels in patients with endometriosis may play a role in the pathogenesis of infertility associated with endometriosis.

Animals↗

Thyroid gland and epididymal function in rats. II. Sperm motile efficiency.

The epididymis is controlled by the endocrine system and plays an active role in the process of spermatic maturation. In order to extend these studies, our investigations were focused on the relationship between the hypofunction of the thyroid gland and the motility of the caput, corpus, and cauda epididymal spermatozoa. The hypofunction was associated with decreasing sperm motility efficiency. In thyroidectomized rats injected with T-3, no sperm motility changes were observed.

Animals↗

Persistent effects of pentoxifylline on human sperm motility, after drug removal, in normozoospermic and asthenozoospermic individuals.

The aim of the study was to compare the in vitro effect of pentoxifylline on human sperm motility when added prior to sperm selection and the persistence of the response after drug removal in normo- and asthenozoospermic individuals. The sperm samples were obtained from 22 men who were repeatedly asthenozoospermic or normozoospermic. Sperm movement was measured using computer-assisted semen analysis over 180 min. Percentage motility and progressive motility were increased in both normo- and asthenozoospermic samples (P < 0.05). Curvilinear velocity, amplitude of lateral head displacement and beat-cross frequency were increased in both groups (P < 0.05). Straight-line velocity was increased significantly in the normozoospermic group only. In both normo- and asthenozoospermic individuals pentoxifylline appeared to enhance sperm motility for at least 180 min after drug removal. This should prevent any potentially toxic effects of the drug on oocytes if it is used to enhance sperm motility during in vitro fertilization.

Humans↗

Factors affecting sperm motility. IX. survival of spermatozoa in various biological media and under different gaseous compositions.

The role of various environmental conditions on sperm motility and their survival was investigated by incubating washed human spermatozoa in various biologic or artificial media and under different atmospheric compositions. When sperm were analyzed by the multiple exposure photography technique for objective motility determination, it was found that none of the investigated biologic media induced any immediate stimulatory effect on the tested spermatozoa. Exchange of seminal fluid between normal and oligoasthenospermic specimens did not reveal any beneficial or harmful effect on sperm motility. Sperm did not survive for more than 24 hours when kept in their original seminal fluid under any circumstances. The most favorable media tested for prolonged sperm survival were pure human umbilical cord serum or Ham's solution containing 0.5% human albumin. However, even in these media sperm did not survive much beyond 24 hours unless incubated in an atmosphere containing 5% CO2 or a triple mixture of 90% N2, 5% O2, and 5% CO2. Only under these circumstances could sperm stay alive, in some cases up to 5 days.

Carbon Dioxide↗

Sperm motility and kinetics of dynein ATPase in astheno- and normozoospermic samples after stimulation with adenosine and its analogues.

We tested the effects of adenosine and 2-deoxyadenosine on the activation of human spermatozoa. In the asthenozoospermic group of patients adenosine produces an increase in sperm motility from 33.3 +/- 2.1% to 42.1 +/- 3.4%, progressive motility from 22.5 +/- 1.3% to 28.6 +/- 1.7% and forward progression rating from 2.1 +/- 0.2% to 2.8 +/- 0.1%. 2-Deoxyadenosine stimulated asthenozoospermic samples to a greater degree than adenosine. Sperm motility rose to 48.9 +/- 3.4%, progressive motility to 32.1 +/- 3.4% and forward progression rating to 3.0 +/- 0.1% following stimulation with 2-deoxy-adenosine. The kinetic parameters and basic characteristics of dynein ATPase were determined. The maximum activity of dynein ATPase, Vmax, was significantly different (P < 0.001) for asthenozoospermic and normozoospermic samples: 6.46 +/- 2.1 nmol Pi/mg/min and 16.99 +/- 3.7 nmol Pi/mg/min respectively. However, the enzyme affinity for ATP was not different. Stimulation of asthenozoospermic samples with adenosine and 2-deoxyadenosine caused an increase of Vmax (70-90% and 90-110% respectively) and no significant change in KM was observed. In order to block the nucleoside transporter and to eliminate the action of adenosine inside the cell, dipyridamole was used but the effects of adenosine were not neutralized. 5'-(N-ethylcarboxy-amido)-adenosine showed effects similar to those of adenosine, even when applied in 1 microM concentration. These results indicate that adenosine and its analogues stimulate sperm motility and activity of dynein ATPase, most probably via A2 receptors.

Adenosine↗

[Significance of the motile sperm presented in the testis].

Generally it is believed that mammalian sperm mature during their transit through the whole epididymis. However spermatozoa aspirated from the epididymal duct or vasa defferentia have been recently reported to move actively in azoospermic patients with seminal tract obstruction. We examined whether the testicular sperm move which would provide useful information in the diagnosis of male infertility. Testicular biopsy materials were obtained from 38 testes of 37 patients as follows; 19 cases with azoospermia, 11 with oligozoospermia, and 8 orchiectomized for prostatic cancer (3), testicular cancer (2), epididymal abscess (1), and cryptorchid (2). All materials were obtained from either open biopsy or from the normal portion of the orchiectomized testis. The material was minced with a sharp knife or scissors in an Eppendorf tube containing Ham's F12 solution. Then a couple of drops of sperm suspension were placed on a warmed (37 degrees C) slide glass which was then covered with a coverglass. The prepared slide was immediately examined by phase-contrast microscopy. Another part was used for preparing a touch smear for confirming the presence of testicular sperm and then was fixed in Bouin's solution and stained with H-E. Spermatogenesis was evaluated by Johnsen's mean score (JMS). Eleven of the 19 azoospermic cases revealed the presence of testicular sperm, and ten of them demonstrated the presence of motile sperm. The mean JMS in these cases was 8.8 (normal spermatogenesis). After surgical exploration or vasography, these patients were diagnosed with obstructive azoospermia (post-vasectomy (4 cases), congenital absence of vas deferens (2 cases), secondary epididymal duct obstruction (4 cases)).(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Effects of experimental cryptorchidism on sperm motility and testicular endocrinology in adult male rats.

The effect of induced cryptorchidism on testicular function and sperm motility was investigated. Bilateral cryptorchidism was created surgically in adult male rats (treated group), and sham-operated rats were used as a control group. Five rats from each group were sacrificed on days 1, 3, 5, and 7 after surgery. The percentage of motile spermatozoa began to decrease 1 day after the operation, followed by an abrupt decline 3 and 5 days later in cryptorchid rats. Furthermore, there were significant decreases in the other sperm motility parameters 5 days after inducement of cryptorchidism. In cryptorchid rats, plasma concentrations of LH, FSH, testosterone, and inhibin B were significantly lower than in the control group 1 day after the operation. Thereafter, plasma concentrations of LH, FSH, and testosterone gradually increased in the cryptorchid rats. On the other hand, plasma concentrations of inhibin B showed a further decline from day 3 after the operation onward. Concentrations of immunoreactive (ir)-inhibin, but not testosterone, in testicular interstitial fluid were remarkably increased until 3 days after surgery in the cryptorchid rats, and declined thereafter. Testicular response to human chorionic gonadotropin (hCG) for testosterone release was decreased in the cryptorchid rats compared with the control rats, indicating that heat stress to testes resulted in a reduction of the activity of Leydig cells and Sertoli cells. These results clearly indicate that heat stress to the testes resulted in a significant reduction of sperm activity within 3 days, and this was followed by changes in testicular endocrine function.

Animals↗

Pentoxifylline stimulates human sperm motility both in vitro and after oral therapy.

Pentoxifylline is a haemorrheologic agent often used in the treatment of peripheral vascular disorders. In this study, we measured sperm motility with a trans-membrane migration method and investigated the effect of this drug in the treatment of male infertility. We found that pentoxifylline increased motility of ejaculated spermatozoa in vitro from both normal and asthenozoospermic samples. After giving pentoxifylline to patients with asthenozoospermia for 3 months, sperm motility significantly increased, but sperm concentration did not increase. From the above results, it can be concluded that pentoxifylline is a useful drug in the treatment of normogonadotropic asthenozoospermia.

Administration, Oral↗

Use of a microcomputer in combination with the multiple exposure photography technique for human sperm motility determination.

An inexpensive microcomputer system that serves as an accessory part of the multiple exposure photography technique for determination of sperm motility is described. With this system the percentage of motile spermatozoa, the average velocity, the frequency distribution of the velocities and the sperm concentration can be computed easily from 200 to 400 images of photographed spermatozoa and within 5 to 8 minutes. In this way the multiple exposure photography technique can be used widely in those centers that are interested in an objective and accurate method for determination of sperm motility in routine semen analysis and for research purposes.

Computers↗

Provocation testing of human sperm motility using energy substrates and activators of the cyclic nucleotide system: II. Studies on sperm from asthenozoospermic subjects.

OBJECTIVE: To find differences in effect on sperm motility of agents that increase intracellular cAMP: manganese ion, methyl-isobutyl-xanthine (MIX), 2-deoxyadenosine, glucose, and Mn-MIX and Mn-glucose. DESIGN: Nine men with asthenozoospermia vs. fertile donors. METHODS: Sperm was washed in Hepes-buffered saline, motility tested by laser-Doppler technique. RESULTS: Best activation was obtained with Mn and 2-deoxyadenosine; generally poor response to MIX or glucose. CONCLUSIONS: Usually, poor endogenous stimulation of adenylyl cyclase, and probably not limited energy supply, is the cause of impaired motility.

1-Methyl-3-isobutylxanthine↗

Poor hypo-osmotic swelling test results from cryopreserved sperm despite preservation of sperm motility.

Cryopreservation thawing of sperm leads to decreased motile density. Most donor programs select for cryopreservation a male with a high initial motile density and if the post-thaw semen has a motile density over 10 x 10(6)/ml, the specimen is thought to represent a fertile specimen. Some recent data suggest that males with normal motile densities but subnormal hypo-osmotic swelling (HOS) tests may be infertile. A study was thus performed to see if males with an adequate motile density after cryopreservation may still demonstrate membrane damage as evidenced by decreased viability and a poor HOS test. The semen species from seven men with motile densities greater than or equal to 10 x 10(6)/ml after freeze-thawing were evaluated for HOS changes and viability. Despite preservation of normal motile density, all 7 men had HOS scores below 50% following cryopreservation (though all were significantly above this level pre-freeze). The mean viability and HOS scores prefreezing were 70 +/- 9.7 and 68.5 +/- 9.5, and post thaw they dropped to 33.7 +/- 6.9 and 32.8 +/- 6.2. These data suggest a mechanism for impaired fertility even with adequate motile density of a thawed specimen that had been cryopreserved.

Cryopreservation↗

Seminal reactive oxygen species and sperm motility and morphology in men with spinal cord injury.

OBJECTIVE: To assess the generation of reactive oxygen species and its relation to semen characteristics in men with spinal cord injury. DESIGN: Cross-sectional study. SETTING: Andrology laboratory at a tertiary care facility and research laboratory at a major medical center. PATIENT(S): Men with spinal cord injury and normal men. INTERVENTION(S): Collecting ejaculates from men with spinal cord injury by electroejaculation and vibratory stimulation and from normal men by masturbation. MAIN OUTCOME MEASURE(S): Measurement of reactive oxygen species before and after stimulation with 50 microM N-formyl-methionyl-leucylphenylalanine (FMLP) and 100 nM 12-myristate 13-acetate phorbol ester (PMA), white blood cell (WBC) concentration, sperm motility and morphology, and ejaculation method. RESULT(S): Compared with controls, levels of reactive oxygen species in men with spinal cord injury were significantly higher in unstimulated, f-MLP-stimulated, and PMA-stimulated specimens. The WBC concentration was significantly elevated in patients with spinal cord injury. Sperm motility in men with spinal cord injury was inversely related to the level of reactive oxygen species. The percentage of morphologically normal spermatozoa was significantly lower in men with spinal cord injury. Levels of seminal reactive oxygen species did not differ when comparing specimen type (antegrade versus retrograde) or method of ejaculation in men with spinal cord injury. CONCLUSION(S): Men with spinal cord injury had elevated levels of reactive oxygen species in their semen. Levels of reactive oxygen species were negatively correlated with sperm motility. Levels of reactive oxygen species were independent of the method of ejaculation or the type of specimen.

Adult↗