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Showing papers on "Semen analysis published in 2002"


Journal ArticleDOI
TL;DR: Sperm morphology, motility, mitochondrial activities and viability are equally susceptible to cryopreservation-induced damage and R123 intensity is a novel and robust indicator of mitochondrial function before and after such trauma.
Abstract: BACKGROUND: The effects of cryoinjury were determined simultaneously on the mitochondrial function, motility, morphology and viability of ejaculated human sperm. METHOD: Rhodamine 123 (R123) uptake (% of sperm) and stain intensity were used to determine sperm mitochondrial activity before and after cryopreservation from the semen of 50 men attending for infertility investigation. Morphology was assessed using Tygerberg’s strict criteria and viability was assessed by eosin Y. Sperm motility was measured using computer-assisted semen analysis (CASA). RESULTS: Freeze–thawing caused a 37% (P 0.001) reduction in normal morphological forms of sperm. All CASA sperm motility parameters except amplitude of lateral head displacement were similarly reduced. R123 uptake and intensity within sperm mitochondria decreased by 36 and 47% respectively (both P 0.001). In addition, there was a similar significant decrease (31%, P 0.001) in the viability of the sperm. CONCLUSIONS: Sperm morphology, motility, mitochondrial activities and viability are equally susceptible to cryopreservationinduced damage. R123 intensity is a novel and robust indicator of mitochondrial function before and after such trauma.

404 citations


Journal ArticleDOI
TL;DR: Infertile men who smoke cigarettes have higher levels of seminal OS than infertile nonsmokers and physicians should advise infertiles men who smoked cigarettes to quit, given the potential adverse effects of seminalOS on fertility.

355 citations


Journal ArticleDOI
TL;DR: Sperm concentration was significantly lower in Columbia, Missouri, than in New York, New York; Minneapolis, Minnesota; and Los Angeles, California, and between-center differences remained significant in multivariate models that controlled for abstinence time, semen analysis time, age, race, smoking, history of sexually transmitted disease, and recent fever.
Abstract: Although geographic variation in semen quality has been reported, this is the first study in the United States to compare semen quality among study centers using standardized methods and strict quality control. We evaluated semen specimens from partners of 512 pregnant women recruited through prenatal clinics in four U.S. cities during 1999-2001; 91% of men provided two specimens. Sperm concentration, semen volume, and motility were determined at the centers, and morphology was assessed at a central laboratory. Study protocols were identical across centers, and quality control was rigorously maintained. Sperm concentration was significantly lower in Columbia, Missouri, than in New York, New York; Minneapolis, Minnesota; and Los Angeles, California. Mean counts were 58.7, 102.9, 98.6, and 80.8 X 10(6)/mL (medians 53.5, 88.5, 81.8, and 64.8 X 10(6)/mL) in Missouri, New York, Minnesota, and California, respectively. The total number of motile sperm was also lower in Missouri than in other centers: 113, 196, 201, and 162 X 10(6) in Missouri, New York, Minnesota, and California, respectively. Semen volume and the percent morphologically normal sperm did not differ appreciably among centers. These between-center differences remained significant in multivariate models that controlled for abstinence time, semen analysis time, age, race, smoking, history of sexually transmitted disease, and recent fever (all p-values < 0.01). Confounding factors and differences in study methods are unlikely to account for the lower semen quality seen in this mid-Missouri population. These data suggest that sperm concentration and motility may be reduced in semirural and agricultural areas relative to more urban and less agriculturally exposed areas.

292 citations


Journal ArticleDOI
TL;DR: Levels of spontaneous and PMA-induced ROS production in pure-sperm suspensions from the infertile men with a diagnosis of leukocytospermia were significantly higher and a similar pattern of increased ROS was observed when spermatozoa were incubated with blood neutrophils.

232 citations


Journal ArticleDOI
TL;DR: Smoking does not affect conventional semen parameters, but significantly increases round cells and leukocytes, and only a few idiopathic infertile smokers were able to quit smoking.
Abstract: BACKGROUND: In this prospective study, semen parameters and hormone concentrations of infertile smokers were compared with infertile non- and ex-smokers. We also determined how many men with idiopathic infertility would stop smoking in an attempt to improve their fertility. METHODS: 1104 men (517 non-smokers, 109 ex-smokers and 478 smokers) with infertility for at least 1 year were evaluated. Evaluation included medical history, physical examination, hormone analysis and two semen analyses. Prior to the second semen analysis, smokers were urged to quit smoking. RESULTS: Smokers were significantly younger (P 1 × 10 6 /ml leukocytes was higher in smokers (P < 0.001). Increased free and total serum testosterone (P < 0.001) and decreased prolactin levels (P < 0.001) were found in smokers. No differences were found between non-smokers and ex-smokers. Only 23.1% of the smokers versus 46% non-smokers (P < 0.001) returned for a second semen analysis, 14 of whom reduced and 15 of whom quit smoking completely. Testosterone levels were significantly lower in those who were able to stop or reduce smoking (P < 0.001). CONCLUSIONS: Smoking does not affect conventional semen parameters, but significantly increases round cells and leukocytes. Only a few idiopathic infertile smokers were able to quit smoking.

205 citations


Journal ArticleDOI
TL;DR: Findings support the hypothesis that there is a genetic basis for variation in postthaw semen quality between individuals, and that AFLP technology may be able to identify molecular markers linked to genes influencing this variation.
Abstract: This study investigated two hypotheses: 1) that consistent between-boar variation in frozen semen quality exists and is genetically determined, and 2) molecular markers linked to genes controlling semen freezability can be identified using amplified restriction fragment length polymorphism (AFLP) technology. Five ejaculates were collected from each of 129 boars. Semen was diluted into a commercial freezing buffer (700 mOsm/kg, 3% glycerol) and five straws (0.5 ml) per ejaculate were cryopreserved (to -5 degrees C at 6 degrees C/min, then -5 degrees C to -80 degrees C at 40 degrees C/min). Semen was assessed for percentage of motile cells, motility characteristics (computer-aided semen analysis; CASA), plasma membrane integrity (SYBR-14 positive), and acrosome integrity (positive for fluorescein-labeled peanut agglutinin; PNA). Consistent between-boar variability was detected for postthaw sperm motility (P 0.05) or straws (P > 0.05) for any viability assessment. Multivariate pattern analysis of the viability data set highlighted three groups of boars producing spermatozoa with poor, average, and good postthaw recovery (42, 63, and 24 boars, respectively). DNA from Large White boars (n = 22) previously classified as good and poor freezers was screened for AFLP markers. Twenty-eight polymerase chain reaction primer combinations generated 2182 restriction fragment bands, of which 421 were polymorphic. Sixteen candidate genetic markers (P < 0.005) were identified by comparing the AFLP profile with semen freezability using logistic regression analysis. These findings support the hypothesis that there is a genetic basis for variation in postthaw semen quality between individuals, and that AFLP technology may be able to identify molecular markers linked to genes influencing this variation.

203 citations


Journal ArticleDOI
TL;DR: The overall high incidence of azoospermia supports the indication for semen cryopreservation in young patients undergoing BMT and underline a need to collect further data through prospective multi-center studies.
Abstract: Knowledge of the impact of different conditioning regimens used in bone marrow transplantation on spermatogenesis is important in pre-BMT counselling for three reasons: (1) Most young patients who have not had children are concerned with their subsequent fertility; (2) For a number of diseases there are competing therapeutic options that may affect spermatogenesis more or less seriously; (3) Since spontaneous recovery of spermatogenesis is rare, it would be necessary to offer cryopreservation as soon as possible after diagnosis and prior to any treatment. This retrospective study evaluates 99 semen samples obtained in 64 patients who underwent BMT between 1982 and 1996. Recovery of spermatogenesis was observed in 90% of patients conditioned with cyclophosphamide (CY), in 50% of patients with CY plus busulphan (BU) or thiotepa and in 17% of patients with CY plus total body irradiation (TBI) or thoracoabdominal irradiation (TAI). Sperm quality following CY was within the normal range (WHO) in the majority of patients, whereas it was consistently severely impaired in patients who received irradiation or two alkylating agents. Following CY, spermatogenesis recovery was observed in 60% of patients tested 1 year post transplant and it was accomplished within the third year in 80% of cases. Following CY + TBI/TAI recovery of spermatogenesis never occurred before the 4th year post transplant and was demonstrated as late as 9 years in one patient who was azoospermic 1 year earlier. No statistical correlation between age and recovery of spermatogenesis could be demonstrated. The overall high incidence of azoospermia (70.3%) supports the indication for semen cryopreservation in young patients undergoing BMT. These results have implications for semen sample timing before and after BMT and underline a need to collect further data through prospective multi-center studies.

174 citations


Journal ArticleDOI
TL;DR: Semen analysis on a male harbouring the A3243G mtDNA mutation is carried out and it is shown that high levels of mutant mtDNA strongly correlate with low sperm motility.
Abstract: Very low levels of somatic mitochondrial (mt)DNA deletions have been identified in the semen of infertile men. It has been suggested that these mutations cause infertility through an effect on sperm motility, but there has been no direct evidence to show that mutant mtDNA can affect sperm function. We have carried out semen analysis on a male harbouring the A3243G mtDNA mutation and show that high levels of mutant mtDNA strongly correlate with low sperm motility.

171 citations


Journal ArticleDOI
TL;DR: Oral Lycopene therapy seem to have a role in the management of idiopathicmale infertility and maximum improvement seems tooccur in the sperm concentration, although largerrandomised controlled trials are essential before definitive therapeutic guidelines can be made.
Abstract: Excessive generation of reactive oxygen species(ROS) containing free oxygen radicals has beenidentified as one of the causes of maleinfertility. Lycopene is a component of humanredox defence mechanism against free radicals.It is found in high concentrations in thetestes and seminal plasma and decreased levelshave been demonstrated in men suffering frominfertility. We evaluated the effect of orallycopene therapy in men with idiopathicinfertility. Beginning March 2000, thirty menwith idiopathic non-obstructiveoligo/astheno/teratozoospermia were enrolledfor the trial. All patients were administered2000 mcg of Lycopene, twice a day for threemonths. Semen analysis was performed at threemonths and sperm concentration, motility andmorphology were evaluated. All patientscompleted the trial without any complications.Twenty patients (66%) showed an improvement insperm concentration, sixteen (53%) hadimproved motility and fourteen (46%) showedimprovement in sperm morphology. In casesshowing an improvement, the median change inconcentration was 22 million/ml, motility 25%and morphology 10%. The improvement inconcentration and motility were statisticallysignificant. Baseline sperm concentration lessthan 5 million/ml was associated with nosignificant improvement. Higher baselineconcentrations were associated with significantimprovement and resulted in six pregnancies in26 patients (23%). Oral Lycopene therapy seemsto have a role in the management of idiopathicmale infertility. Maximum improvement seems tooccur in the sperm concentration (66% cases).Patients without severe oligospermia (spermdensity >5 million/ml) may be given a trialof therapy with lycopene. However, largerrandomised controlled trials are essentialbefore definitive therapeutic guidelines can bemade.

158 citations


Journal ArticleDOI
TL;DR: It is found that sperm motility was reduced significantly by H2O2 concentrations 20-fold lower in EYTG than in TALP medium, and the addition of low amounts of catalase and millimolar concentrations of pyruvate greatly improved the antioxidant properties of a commonly used extender.

154 citations


Journal ArticleDOI
TL;DR: Semen evaluation in a large population of HIV-infected men requesting ART evidenced several alterations, some of which might be related to anti-retroviral treatments.
Abstract: BACKGROUND Couples in whom the man is infected by human immunodeficiency virus (HIV) increasingly request assisted reproductive technology (ART) to allow safe procreation. Semen quality is critical in such situations. METHODS Semen characteristics were evaluated in 189 HIV-infected men requesting ART. At the time of semen analysis all men were healthy and 177 were receiving anti-retroviral therapy. Comparisons were made with HIV-seronegative men, partners of women requiring IVF because of tubal infertility, after matching for age and sexual abstinence delay. RESULTS The most significant semen alterations found in the HIV-infected men were reduced percentages of rapidly progressive sperm [median (range), 10% (0-30%) compared with 15% (5-30%) in the controls, P < 0.001], and increased concentrations of non-spermatic cells [3 x 10(6)/ml (0.2-16 x 10(6)/ml) compared with 1.1 x 10(6)/ml (0.1-14 x 10(6)/ml) in the controls, P < 0.001]. HIV-infected men also showed lower ejaculate volumes [2.8 ml (0.6-9.3 ml) compared with 3.6 ml (1.1-11 ml), P < 0.05] and total sperm counts [262.5 x 10(6) (0-1003 x 10(6)) compared with 310.5 x 10(6) (48.3-1679 x 10(6)), P < 0.05]. CONCLUSIONS Semen evaluation in a large population of HIV-infected men requesting ART evidenced several alterations. Some of these anomalies might be related to anti-retroviral treatments.

Journal ArticleDOI
01 Dec 2002-Urology
TL;DR: The data show that sperm DD negatively correlates with standard semen parameters and that an isolated abnormality of sperm DD, a marker of sperm DNA integrity, is uncommon in infertile men.

Journal ArticleDOI
TL;DR: The results indicate that the infertility of the male Smcp−/− mice on the 129/Sv background is due to reduced motility of the spermatozoa and decreased capability of the semen analysis system to penetrate oocytes.
Abstract: The sperm mitochondria-associated cysteine-rich protein (SMCP) is a cysteine- and proline-rich structural protein that is closely associated with the keratinous capsules of sperm mitochondria in the mitochondrial sheath surrounding the outer dense fibers and axoneme. To investigate the function of SMCP, we generated mice with a targeted disruption of the gene Smcp by homologous recombination. Homozygous mutant males on a mixed genetic background (C57BL/6J × 129/Sv) are fully fertile, while they are infertile on the 129/Sv background, although spermatogenesis and mating are normal. Homozygous Smcp−/− female mice are fertile on both genetic backgrounds. Electron microscopical examination demonstrated normal structures of sperm head, mitochondria, and tail. In vivo experiments with sperm of Smcp−/− 129/Sv mice revealed that the migration of spermatozoa from the uterus into the oviduct is reduced. This result is supported by the observation that sperm motility as determined by the computer-assisted semen analysis system (CASA) is significantly affected as compared to wild-type spermatozoa. In vitro fertilization assays showed that Smcp-deficient spermatozoa are able to bind to the oocyte but that the number of fertilized eggs is reduced by more than threefold relative to the wild-type control. However, removal of the zona pellucida resulted in an unaffected sperm-egg fusion which was monitored by the presence of pronuclei and generation of blastocyts. These results indicate that the infertility of the male Smcp−/− mice on the 129/Sv background is due to reduced motility of the spermatozoa and decreased capability of the spermatozoa to penetrate oocytes.

Journal ArticleDOI
TL;DR: Comparisons of sperm counts in a population of Danish military conscripts and a corresponding cohort from Southern Sweden found differences in reproductive parameters could not be explained by possible confounders and may be due to unknown environmental or lifestyle-related factors.
Abstract: BACKGROUND: A common aetiology behind time-related deterioration of male reproductive function, including decline of sperm concentration and rising incidence of testicular cancer and cryptorchidism, has been suggested. Accordingly, a high incidence of testicular cancer and low sperm counts have been observed in Denmark, while in Finland opposite figures were found. This may be due to genetic factors. The incidence of testicular cancer is 50% lower in Sweden than in Denmark. Data on sperm counts in a population of Danish military conscripts have been published and we wished therefore to compare them with semen parameters in a corresponding cohort from Southern Sweden, geographically and genetically very closely related to Denmark. METHODS: A total of 305 military conscripts was recruited and investigated in an identical way as those in the Danish study including a questionnaire, physical examination and semen analysis. RESULTS: Men born and raised in Sweden had a 23% higher mean sperm concentration, a 31% higher mean total sperm count and a 14% higher seminal volume compared with their Danish counterparts. CONCLUSIONS: The differences in reproductive parameters between the two genetically very similar populations could not be explained by possible confounders and may be due to unknown environmental or lifestyle-related factors.

Journal ArticleDOI
TL;DR: It is demonstrated that 5 x 10(6) activated neutrophils/ml are sufficient to impair equine sperm motility in vitro, and this effect was associated with a significant decrease in total motility.

Journal ArticleDOI
TL;DR: The findings of elevated seminal plasma oestradiol, decreased testosterone and testosterone/oestradio ratio in infertile men, and the significant correlation between hormone levels and sperm analysis data suggest that the local balance between androgen and oestrogen is important for spermatogenesis.
Abstract: The hypothesis that the balance between oestrogen and androgen in seminal plasma is important for normal fertility was investigated. We determined the concentrations of oestradiol and testosterone in blood and seminal plasma from 62 infertile men and 32 normozoospermic men. Infertile men were classified according to semen analysis (concentration, motility and morphology): asthenozoospermia, oligozoospermia and oligoteratoasthenozoospermia. Serum luteinizing hormone (LH) and follicle-stimulating hormone (FSH) were determined in all participants. For all subjects, mean testosterone levels were lower and mean oestradiol were higher in seminal plasma than in blood. Seminal plasma testosterone levels were lower in the infertile groups vs. control men ( p < 0.0002). Oligpzoospermic and oligoteratoasthenozoospermic men had significantly higher seminal plasma oestradiol levels compared with controls ( p < 0.03). The three infertile groups had significantly lower seminal plasma testosterone/oestradiol ratio than control men ( p < 0.001). Sperm analysis data (concentration, motility and morphology) significantly correlated with seminal plasma testosterone/oestradiol ratio. The findings of elevated seminal plasma oestradiol, decreased testosterone and testosterone/oestradiol ratio in infertile men, and the significant correlation between hormone levels and sperm analysis data suggest that the local balance between androgen and oestrogen is important for spermatogenesis.

Journal ArticleDOI
TL;DR: The results of this study indicate that male patients with hyperthyroidism have abnormalities in seminal parameters, mainly sperm motility, which improve or normalize when the patients become euthyroid.
Abstract: The aim of this prospective controlled study was to ascertain the effect of hyperthyroidism on sperm quality and composition. We studied 23 thyrotoxic male patients, aged 43.8 +/- 2.4 yr (mean +/- SEM), and 15 healthy male controls of approximately the same age (42.2 +/- 2.2 yr). Two semen analyses at intervals of 2-3 wk were obtained before and about 5 months after euthyroidism was achieved either by methimazole alone (14 patients) or (131)I plus methimazole (9 patients). Total fructose, zinc (Zn), and magnesium (Mg) were also measured in seminal plasma in 16 patients, because 7 had semen volume less than 2 ml. In the control group semen analysis was performed only once. Mean (+/-SEM) semen volume was within normal range both in patients (3.3 +/- 0.2 ml) and controls (3.5 +/- 0.4 ml; P = NS). Mean sperm density was lower in patients, although the difference compared with controls did not reach statistical significance (35.7 +/- 5.3 vs. 51.5 +/- 6.1 x 10(6)/ml; P = 0.062). The same was found with sperm morphology (68 +/- 7% vs. 78 +/- 8%; P = NS). Finally, mean motility was lower in thyrotoxic males than in controls (28 +/- 8% vs. 57 +/- 7%; P < 0.01). After treatment, sperm density and motility improved [35.7 +/- 5.3 vs. 43.3 +/- 6.5 x 10(6)/ml (P = NS) and 28 +/- 8% vs. 45 +/- 7% (P < 0.05), respectively], but sperm morphology did not change (68 +/- 7% vs. 70 +/- 6%; P = NS). Mean values for fructose, Zn, and Mg did not differ between controls and patients either before or after achievement of euthyroidism [9.2 +/- 0.7, 3.0 +/- 0.5, and 4.2 +/- 0.7 nmol/liter vs. 8.6 +/- 0.9, 3.0 +/- 0.5, and 4.7 +/- 0.8 nmol/liter (patients before) and 9.1 +/- 0.7, 3.1 +/- 0.6, and 4.5 +/- 0.9 nmol/liter (patients after treatment) for fructose, Zn, and Mg, respectively]. Moreover, according to the treatment given, no statistically significant differences were found before or after treatment. Finally, seminal plasma fructose, Zn, and Mg levels did not correlate with sperm parameters or with pretreatment thyroid hormone levels. In conclusion, the results of our study indicate that male patients with hyperthyroidism have abnormalities in seminal parameters, mainly sperm motility. These abnormalities improve or normalize when the patients become euthyroid. Restoration of sperm parameters was independent of the treatment provided for the hyperthyroid syndrome. Moreover, seminal plasma elements, such as fructose, Zn, and Mg, did not correlate with sperm density, motility, or morphology.

Journal ArticleDOI
TL;DR: It is demonstrated that approximately one-quarter of men utilising cryopreserved semen after treatment for Hodgkin's disease obtain a live birth and the high non-utilisation rate is intriguing and warrants further investigation.
Abstract: Between 1978 and 1990, 122 men underwent semen analysis before starting sterilising chemotherapy for Hodgkin's disease. Eighty-one (66%) had semen quality within the normal range, 25 were oligospermic (<20×106 sperm per ml) and five were azoospermic (no sperm in the ejaculate). Semen from 115 men was cryopreserved and after a median follow-up time of 10.1 years, 33 men have utilised stored semen (actuarial rate 27%) and nine partners have become pregnant resulting in 11 live births and one termination for foetal malformation. Actuarial 10 year rates of destruction of semen before death or utilisation and death before utilisation are 19% and 13% respectively. This retrospective cohort study demonstrates that approximately one-quarter of men utilising cryopreserved semen after treatment for Hodgkin's disease obtain a live birth. The high non-utilisation rate is intriguing and warrants further investigation. British Journal of Cancer (2002) 87, 381–384. doi:10.1038/sj.bjc.6600483 www.bjcancer.com © 2002 Cancer Research UK

Journal ArticleDOI
TL;DR: The causes for the spermatogenetic defects in men with idiopathic infertility, a history of maldescended testes or varicocele are acquired before adolescence and do not progress during adulthood, and genital tract infections may be suspected as a contributing factor.
Abstract: Summary. In a cross-sectional retrospective study, the age-dependency of semen parameters of men with specific andrological diseases was investigated in 3698 men (mean age 33 years; range 19–63 years) attending a tertiary referral clinic for couple infertility. In all, 1853 patients suffered from idiopathic infertility (mean age 33.2 years; range 19–60 years), 388 from infections of the accessory glands (34.2; 20–61), 363 had a history of maldescended testes (32.0; 20–51) and 833 a varicocele (33.6; 20–63). In 172 patients, a varicocele and an infection (34.3; 24–54) and in 94 a varicocele and a history of maldecended testes (32.5; 21–53) were found. Semen analysis was performed according to WHO criteria. An age-related decrease in testicular volume was not observed in any group. Total sperm count decreased significantly with advancing age only in patients with an infection of the accessory glands. We found no evidence for age-dependent progressive deterioration of semen variables in the other groups. It may be concluded that the causes for the spermatogenetic defects in men with idiopathic infertility, a history of maldescended testes or varicocele are acquired before adolescence and do not progress during adulthood. In contrast, infections of the accessory glands may lead to decreased sperm counts by functional impairment or partial occlusion of the seminal ducts. Thus, if in the general population declining sperm counts are observed, genital tract infections may be suspected as a contributing factor.

Journal ArticleDOI
TL;DR: The Comet assay proved to be sensitive in detecting an alteration in DNA integrity in germ cells of workers exposed to styrene, contributing towards the understanding of the importance of male occupational exposure within the context of genetic risk assessment in humans.
Abstract: BACKGROUND: Occupational exposure to toxic agents may cause infertility, congenital anomalies or death in offspring, but few studies have evaluated DNA integrity in germ cells of male workers. We investigated sperm DNA integrity in individuals occupationally exposed to styrene. METHODS AND RESULTS: Semen samples were obtained from 46 male workers exposed to styrene and 27 unexposed controls (age range 18-45 years). Exposed individuals had worked for at least 2 years in the last 5 years and continuously for 6 months in factories producing reinforced plastics. The Comet assay was performed to evaluate DNA integrity in sperm, as well as semen quality analysis to assess sperm concentration and morphology. There were no differences in the results of the standard semen analysis between exposed subjects and the reference group. However, we found a significant difference (P < 0.001) in sperm DNA damage by the Comet assay between exposed subjects and the reference group. CONCLUSIONS: The Comet assay proved to be sensitive in detecting an alteration in DNA integrity in germ cells of workers exposed to styrene. This finding contributes towards the understanding of the importance of male occupational exposure within the context of genetic risk assessment in humans.

Journal Article
TL;DR: In this paper, the authors performed extensive studies on relationship between sperm functions and fertilization rates by logistic regression analysis in large numbers of IVF patients including 370 patients with zero fertilization rate by IVF.
Abstract: Evaluation and assessment of semen is very important for both diagnosis of male infertility and selection of patients for treatment with IVF or ICSI. In standard IVF, sperm function is essential for normal fertilization: sperm must be able to bind to zona pellucida (ZP), undergo the acrosome reaction and penetrate the ZP and fuse with the oolemma before fertilization takes place. In contrast, most sperm functions are not required for fertilization in ICSI since sperm bypass the ZP and oolemma by injection of a single sperm directly into cytoplasm of oocyte. Therefore, the clinical decision on treatment of patients with either IVF or ICSI is mostly dependent on results of sperm tests. However, conventional semen analyses do not provide accurate information about sperm fertilizing ability since many patients with subtle sperm defects can not be detected. More advanced sperm function tests are required to detect sperm defects that may lead to failure of fertilization in standard IVF. In the last 15 years we performed extensive studies on relationship between sperm functions and fertilization rates by logistic regression analysis in large numbers of IVF patients including 370 patients with zero fertilization rate by IVF. We confirmed sperm morphology assessed strictly was strongly related to fertilisation rate with standard IVF. Thus sperm morphology assessment is very useful for selection of patients for ICSI. We also developed a number of new tests including sperm-ZP binding, sperm-ZP penetration and the ZP-induced AR and evaluated the clinical value of these tests. Sperm-ZP binding and sperm-ZP penetration tests are the most powerful indicators for sperm fertilizing ability in vitro. The ZP-induced AR is highly correlated with sperm-ZP penetration. We discovered a condition we call disordered ZP-induced AR which causes serve infertility in up to 25% men with otherwise idiopathic infertility In conclusion, the combination of semen analysis with advanced sperm function tests provide important diagnostic and prognostic information for male infertility and is crucial for selection of patients for treatment with IVF or ICSI.

Journal ArticleDOI
TL;DR: Motility is not a good enough criterion on which to prove the presence of viral elements, either in the medium or on the sperm, as it depends on the type of virus present as well as the presence or absence of seminal fluid.
Abstract: Background The effect of viral particles on the motility of human sperm and the relationship between sperm and virus are of importance particularly in assisted fertilization. Methods We incubated ejaculated sperm with or without seminal fluid with either herpes simplex virus type 2 (HSV2) or human cytomegalovirus (HCMV). For each experiment, 5 x 10(5) sperm were incubated with a viral load of between 10(4) and 10(6) plaque-forming units. Results We detected no apparent variations in the percentage of motile forms when sperm were incubated with either HSV2 or HCMV. Using a computer-aided semen analysis system, a slight difference was reported in the percentage of motile forms when seminal fluid-free sperm were incubated with HSV2 (57.18 versus 64.43 in the control). Although the mean amplitude of lateral head displacement and the curvilinear velocity were significantly higher in infected sperm, the difference in straight line velocity was not statistically significantly different. Few viral particles (HSV2 or HCMV) adhered to the sperm membrane in the presence of seminal fluid. However, more particles stuck when in the absence of seminal fluid, particularly with HSV2 (8% of sperm sections for HSV2; 4% for HCMV). Conclusions The relationship between sperm and viruses depends on the type of virus present as well as the presence or absence of seminal fluid. Motility is not a good enough criterion on which to prove the presence of viral elements, either in the medium or on the sperm.


01 Jan 2002
TL;DR: Evaluation of the quality of cryopreserved bull spermatozoa is reviewed and simultaneous analysis of multiple sperm attributes, or outcomes of several laboratory assessments must be combined to look for the overall effect of several independent sperm parameters.
Abstract: Evaluation of the quality of cryopreserved bull spermatozoa is reviewed. The methods for the assessment of sperm quality have markedly improved over the last decades - starting with the assessment of morphological shapes and subjective motility analysis towards the more sophisticated analysis of the molecular changes in chromatin, membranes and catabolic activities of the sperm cell itself. Function of sperm plasma membrane under the hypo- osmotic conditions, distribution and concentration of ions, or function of different organelles seem to correlate with the degree of the viability of spermatozoa after freezing and thawing procedures. Many in vitro techniques that stimulate sperm function through female-derived factors, such as zona and oocyte, in zona pellucida binding assay, in vitro fertilization (IVF) and production of embryos, were employed to predict the outcome of artificial insemination in the field. Still, majority of methods used for semen analysis today are both - tedious and expensive, and, in many cases confined to human bias. In order to increase the predictive power of assessment, simultaneous analysis of multiple sperm attributes, or outcomes of several laboratory assessments must be combined to look for the overall effect of several independent sperm parameters.

Journal ArticleDOI
TL;DR: HPG axis suppression with MPA-testosterone for > or =12 weeks did not restore spermatogenesis in azoospermic men treated with gonadotoxic radiotherapy and chemotherapy for childhood cancer.
Abstract: BACKGROUND Does suppression of the hypothalamic-pituitary-gonadal (HPG) axis restore spermatogenesis in men rendered azoospermic following treatment of childhood cancer? METHODS Seven men with azoospermia secondary to treatment for childhood cancer, median age (range), 22.2 (18-25.3) years, aged 10.4 (4.4-13.3) years at original diagnosis, participated. Each subject underwent semen analysis and testicular biopsy, followed by treatment with medroxyprogesterone acetate (MPA), 300 mg i.m. repeated after 12 weeks, with 800 mg testosterone pellets s.c. on day 1 to suppress the HPG axis. Hormone and semen analysis was performed every 6 weeks for 48 weeks. A second testicular biopsy was performed at week 48. RESULTS Before HPG axis suppression, mean +/- SEM plasma LH was 9.0 +/- 1.8 U/l, testosterone 17.9 +/- 1.5 nmol/l and FSH 22.4 +/- 4.4 U/l. Median (range) venous plasma and seminal plasma inhibin B levels were 10.0 (7.8-35) and 11.2 (7.8-770) ng/l respectively. During HPG suppression, FSH and LH levels were undetectable for > or =12 weeks followed by a gradual return to pretreatment concentrations by 48 weeks. All men remained azoospermic at study completion and complete absence of germ cells on biopsies was demonstrated by immunocytochemistry for all specimens pre- and post-HPG axis suppression. CONCLUSIONS HPG axis suppression with MPA-testosterone for > or =12 weeks did not restore spermatogenesis in azoospermic men treated with gonadotoxic radiotherapy and chemotherapy for childhood cancer.

Journal ArticleDOI
TL;DR: Results indicate that in normospermic men seminal plasma Estradiol levels are higher than blood hormone levels, suggesting local production of estradiol, which may imply that estrogen and/or the balance andorgen/estrogen is important in normal human spermatogenesis.
Abstract: The authors determined semen quality and the concentrations of estradiol, testosterone, and melatonin in blood and seminal plasma of 8 normal men. To investigate the reproducibility of these parameters, semen analysis and hormone concentrations were determined on 3 occasions, 6 weeks apart. All 8 men had normal semen analysis. Blood melatonin (9.7-45.4 pg/mL) and testosterone (3.5-12.3 ng/mL) levels were significantly higher than the comparable seminal plasma levels (0.6-5.0 pg/mL, p <.02; 0.1-0.9 ng/mL, p <.0001, respectively). Seminal plasma estradiol levels (46.9-91.3 pg/mL) were significantly higher than the blood levels (13.3-44.7 pg/mL) (p <.0001). The intraindividual variations in seminal plasma estradiol levels ranged between 8.7 and 13.8%. There was no correlation between sperm concentration, motility or morphology and blood or seminal plasma hormone levels. Also, blood and seminal plasma hormone levels were not correlated. These results indicate that in normospermic men seminal plasma estradiol levels are higher than blood hormone levels, suggesting local production of estradiol. This may imply that estrogen and/or the balance andorgen/estrogen is important in normal human spermatogenesis.

Journal ArticleDOI
TL;DR: This study shows that the outcome of fresh and frozen–thawed testicular spermatozoa in ICSI is comparable, obstructive and nonobstructive etiologies perform the same, and that preincubation of testicular semen results in increased fertilization and pregnancy rates.
Abstract: Purpose: The aim of the study was to gain an insight into the optimal management of the infertile couple with the husband suffering from azoospermia.

Journal ArticleDOI
TL;DR: Several assays have been developed that are useful for assessing sperm DNA composition and organization and revealed that when sperm DNA has been damaged or packaged improperly there is a concomitant and often significant decline in male fertility.
Abstract: Traditional semen analysis is essential for the diagnosis of male infertility. A number of studies over the past decade have reported that a significant contributing factor to male fertility that is not revealed as part of semen analysis is sperm DNA, specifically its composition and organization. Exogenous and endogenous factors can cause damage to sperm DNA. For example, topoisomerase II activity, which is necessary for sperm DNA packaging, can adversely influence the competence of sperm DNA if the activity of the enzyme is abnormal. Germ cell apoptosis can be induced by oxygen radicals produced from environmental (for example cigarette smoke) or testicular (for example localized ischaemia) sources. Several assays have been developed that are useful for assessing sperm DNA composition and organization. To date, each of these assays has revealed that when sperm DNA has been damaged or packaged improperly there is a concomitant and often significant decline in male fertility.

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TL;DR: The combined presence of IgA Ab to human 60 kDa HSP, leukocytes and other established infection/inflammation markers in semen might suggest a potential role of the immune response to heat shock proteins (HSP) in cases of silent male genital tract infection, but the results do not indicate a marked relationship of HSP 60 Ab in seminal fluid with standard parameters of semen quality.
Abstract: Background Heat shock proteins (HSP) are essential mammalian and bacterial stress proteins. At the cellular level, they act as chaperones, have important regulatory functions, and are considered to be an essential factor for reproduction. Scarce information exists on the role of sensitization to HSP and the potential role in the aetiology of male infertility. Methods The potential association of immunoglobulin (Ig)A antibodies (Ab) to the human 60 kDa heat shock protein (HSP 60) with several parameters of subclinical male genital tract infection/inflammation and with semen quality and sperm fertilizing capacity was analysed in a prospective study. IgA Ab to human HSP 60 were determined in seminal plasma of 202 randomly chosen male partners of subfertile couples with a median duration of infertility of 4 years (range 1-15 years), who were asymptomatic for genital tract infection. After medical history and clinical examination, a comprehensive evaluation of semen quality, in aliquots of the same ejaculates used for HSP Ab determination, included: sperm analysis; local antisperm antibody (ASA) screening; standardized sperm-cervical mucus (CM) penetration testing; immunocytochemical round cell differentiation to determine seminal leukocyte counts; evaluation of complement fraction C(3) and of some pro-inflammatory cytokines; and microbial screening. Subsequent fertility was recorded after 6 months. Results The presence of HSP 60 IgA Ab in seminal fluid (total positive 6.9%) was significantly associated with leukocytospermia, the presence of C(3), and also with high interleukin (IL) levels in seminal plasma. HSP 60 Ab were not related to the bacterial colonization of ejaculates. There was no association of seminal IgA Ab to human HSP 60 with semen quality, determined with microscopical semen analysis, nor with local IgG- or IgA-class ASA. There was no relationship with sperm intrinsic motility and duration of motility in the sperm CM-penetration test, nor with sperm fertilizing capacity. Conclusions The combined presence of IgA Ab to human 60 kDa HSP, leukocytes and other established infection/inflammation markers in semen might suggest a potential role of the immune response to heat shock proteins (HSP) in cases of silent male genital tract infection, but the results do not indicate a marked relationship of HSP 60 Ab in seminal fluid with standard parameters of semen quality.

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TL;DR: DFS sperm have increased cross-reactivity to anti-ubiquitin antibodies, a finding consistent with the ubiquitination of defective sperm shown in animal models, which justifies the use of ubiquitin-based assays for objective semen analysis in infertile men with heritable defects.
Abstract: BACKGROUND: Human sperm with structural abnormalities display an increased content of the cellular proteolytic marker peptide, ubiquitin. We investigated whether dysplasia of the fibrous sheath (DFS), a severe structural anomaly found in the sperm of some asthenozoospermic patients, is accompanied by (i) increased ubiquitination of the sperm surface and (ii) by increased ubiquitination of the sperm mitochondria. METHODS AND RESULTS: Five DFS patients and eight fertile donors were studied by immunocytochemistry with anti-ubiquitin antibodies. Increased cross-reactivity of the ubiquitinated mitochondrial epitopes was seen in 32–50% of DFS sperm, but only 2–4.1% of sperm from fertile donors. Sperm surface ubiquitination assessed by sperm-ubiquitin tag immunoassay (SUTI) and immunofluorescence demonstrated an increased sperm ubiquitination in all DFS patients. The average median value of ubiquitin-induced fluorescence in DFS patients was 25.8 counts (range 19.8–37.9), as opposed to 13.4 counts range (9.3–16.6) in fertile men. Sperm with ‘stump tails’, coiled tails, twin and triplet sperm, and clusters of immature spermatogenic cells were common. CONCLUSIONS: DFS sperm have increased crossreactivity to anti-ubiquitin antibodies, a finding consistent with the ubiquitination of defective sperm shown in animal models. These results justify the use of ubiquitin-based assays for objective semen analysis in infertile men with heritable defects.