Normal Sperm Concentration Research Articles

Abnormally high body mass index and tobacco use are associated with poor sperm quality as revealed by reduced sperm binding to hyaluronan-coated slides

Responses on a lifestyle questionnaire were correlated with results from traditional semen analysis and a newer functional sperm assay, namely, the ability of sperm to bind to a hyaluronan-coated slide. Increased percent normal morphology and motile sperm concentration were positively correlated with increased HA-binding score, whereas tobacco use (either current or previous history) and abnormally high body mass index were associated with lower binding scores.

Impact of chemotherapeutics and advanced testicular cancer or Hodgkin lymphoma on sperm deoxyribonucleic acid integrity

To determine the impact of combination chemotherapy on the quality of newly generated spermatozoa from patients with advanced testicular cancer and patients with Hodgkin lymphoma (HL). Prospective longitudinal study. Academic facility. Patients with newly diagnosed metastatic testicular cancer and with HL that required chemotherapy were compared with age-matched healthy community volunteers. None. Semen parameters, hormone levels, testis volume, and presence of sperm DNA strand breaks in patients with cancer and in healthy community volunteers were compared before and after the patients' chemotherapy at 6, 12, 18, and 24 months. Before chemotherapy, both cancer groups had poorer semen quality compared with community volunteers. Among patients with testicular cancer and HL, 67% and 60%, respectively, had <5x10(6) sperm/mL at 6 months after chemotherapy. At 24 months, 60% and 57% of patients with testicular cancer and HL, respectively, had normal sperm concentrations. Level of FSH was significantly higher in the cancer group compared with community volunteers at 6 to 12 months after chemotherapy. Before chemotherapy, sperm DNA damage was higher in the cancer group than in community volunteers; this damage was increased further at 6 months and remained elevated 24 months after treatment. Sperm generated after chemotherapy maintain a significant degree of chromatin damage. Therefore, survivors of testicular cancer and HL are at risk of having abnormal reproductive outcome. Proper counseling to these patients on reproductive risks and fertility preservation before chemotherapy is recommended.

Assessment of sperm factors possibly involved in early recurrent pregnancy loss

To evaluate and compare standard sperm parameters, lipid peroxidation of sperm plasma membranes, antioxidant capacity of seminal plasma, and sperm chromatin integrity in ejaculates from men whose partners have a history of recurrent pregnancy loss and from a control group of men with recent fertility. Descriptive study. Reproduction Program, University of Antioquia, Medellín, Colombia. Twenty-three couples with history of recurrent pregnancy loss and 11 men with recent fertility. Semen samples from control men and men whose partner had a history of recurrent pregnancy loss were examined for differences in semen parameters, lipid peroxidation, antioxidant capacity, DNA fragmentation index, and sperm preparation. Migration-sedimentation method was used to collect motile spermatozoa from fresh ejaculates for examination for semen parameters. Men from the control group had spermatozoa with higher percentage of normal sperm morphology, concentration, progressive motility, and antioxidant capacity compared with men from the recurrent pregnancy loss group, who had spermatozoa with higher teratozoospermia and higher lipid peroxidation. Motile sperm fractions from both groups had spermatozoa with better sperm parameters compared with freshly ejaculated sperm. This study strengthens the current literature associating sperm quality with recurrent pregnancy loss, and emphasizes the importance of evaluating male factor by tests such as lipid peroxidation and measuring antioxidant capacity of seminal plasma in addition to conventional sperm parameters.

Evidence of sperm nuclear chromatin heterogeneity in ex-cryptorchid subjects.

Ex-cryptorchid subjects are frequently affected by infertility, even if the usually determined seminal parameters are still within the normal range. We evaluated in this study, in 14 ex-unilateral cryptorchid adult males (22-28 years), with normal sperm concentration, morphology and viability, the resistance of sperm nuclear chromatin to denaturation, using the fluorochrome acridine orange, which fluorescences green when bound to native DNA (double stranded) and red when bound to denaturated DNA (single stranded). Our findings demonstrate that the percentage of green-stained cells with acridine orange was significantly lower in our patients than in controls (p less than 0.001). We therefore suggest that the chromatin of ex-cryptorchid subjects' spermatozoa has a decreased resistance to denaturation and we hypothesize that this finding may explain the infertility of these patients, when the common seminal parameters are still within the normal range.

Follicle-stimulating hormone autoantibody is involved in idiopathic spermatogenic dysfunction

To detect the anti-follicle-stimulating hormone (FSH) antibody in idiopathic infertile patients and fertile subjects in order to determine the role of this antibody in patients with spermatogenic dysfunction. The anti-FSH antibody in serum was detected by an enzyme-linked immunosorbent assay (ELISA). The functional and structural integrity of the sperm membrane was evaluated with hypo-osmotic swelling (HOS) test and the ultrastructure of the spermatozoa was investigated by transmission electron microscopy (TEM). The extent of positive FSH antibody in the patients with oligozoospermia and/or asthenozoospermia was significantly higher than that in the fertile subjects and infertile patients with normal sperm concentration and motility, but it was significantly lower than that in the patients with azoospermia. The extent of anti-FSH antibody in the patients with azoospermia was significantly greater than that in patients with oligospermia and/or asthenospermia, infertile people with normal sperm density and motility and fertile people. The hypo-osmotic swelling test showed that the percentage of HOS-positive spermatozoa (swollen) was 45.1 mu 3.5% in the FSH antibody-positive group and 59.1% micro 6.2% in the FSH antibody-negative control group. The percentage of functional membrane damage to spermatozoa was significantly higher in the anti-FSH antibody-positive group than in the control group. TEM showed that the outer acrosomal membrane was located far from the nucleus, and detachment of the acrosome was found in the FSH autoantibody-positive group. These data suggest that the presence of anti-FSH antibody is strongly correlated with the sperm quantity and quality in idiopathic male infertility. Anti-FSH antibody may be an important factor causing spermatogenic dysfunction and infertility.

Sperm ultrastructure and meiotic segregation in a group of patients with chronic hepatitis B and C

Little is known about the effect of chronic hepatitis B and hepatitis C on sperm quality. In this study, we analysed sperm quality from selected patients with chronic hepatitis B virus (HBV) or hepatitis C virus (HCV) infections. Semen samples were examined by light and transmission electron microscopy (TEM). TEM data were elaborated with a mathematical formula able to indicate a fertility index and the presence of the three main sperm pathologies: apoptosis, immaturity and necrosis. Meiotic chromosome segregation was investigated by fluorescence in situ hybridisation carried out on sperm nuclei, using probes for chromosomes 18, X and Y. Despite normal sperm concentration, we observed reduced motility. TEM analysis highlighted that 35.7% of patients showed generally good semen quality. However, significantly higher values of apoptosis and necrosis, compared with controls, were observed, demonstrating spermatogenetic alterations. Regarding meiotic segregation, we found an incidence of disomies similar to that observed in control samples, whereas diploidy resulted higher in HCV patients, without reaching statistical significance. In conclusion, sperm quality in the studied group was not impaired; however, apoptosis and necrosis resulted out of normal range and the fertility index was significantly lower in HCV- and HBV-infected patients versus controls.

Sperm cryopreservation in patients with hematologic malignancies

Many patients with acute leukemia or lymphoma are cured by chemotherapy or conditioning followed by hemopoietic stem cell transplantation. However, cancer chemotherapies and conditioning regimens deteriorate spermatogenesis. The greatest risk of infertility caused by chemotherapy is associated with alkylating agents such as cyclophosphamide, busulfan and melphalan, which are frequently used as conditioning in hemopoietic stem cell transplantation [1, 2]. Cyclophosphamide is the key drug for treatment of nonHodgkin lymphoma. Non-alkylating drugs such as anthracylines and cytarabine have been shown to have less effect or only a transient adverse effect on sperm production in children with acute leukemia [3, 4]. However, there are few reports on sperm production in adults with acute leukemia. Recently, Bahadur et al. [5] showed that gonadal toxic treatment for adult patients with leukemia or lymphoma results in significant reduction in sperm quality. Similar results were reported by Lemez and Urbanek [6]. Obviously, the best way to rescue infertility in adult male patients with leukemia, lymphoma or related disorders is sperm cryopreservation before the start of chemotherapy [7, 8]. We previously reported the results of a preliminary study showing that it is difficult to obtain the normal number of sperms in heavily treated patients with hematologic malignancies [9]. In this report, we summarize data on sperm collected for cryopreservation from patients with hematologic malignancies and describe successful spouse pregnancy using cryopreserved sperm from a patient with acute myeloid leukemia (AML). Before the start of initial chemotherapy, patients were informed of adverse effects related to chemotherapy, including mucositis, diarrhea, infections, secondary malignancies, and infertility. When male patients expressed a desire to cryopreserve their sperm, they were referred to private clinics conducting sperm banking. Data on sperm concentration and motility before cryopreservation were obtained from each clinic by letters in reply. From January 1999 to December 2007, sperm cryopreservation was performed for 38 patients with hematologic malignancies. Data on sperm from six patients previously reported [9] were included. The patients were divided into two groups: one group of the patients whose sperm was cryopreserved before the start of initial chemotherapy and another group of the patients whose sperm was cryopreserved after chemotherapy had been undertaken. Table 1 shows a summary of sperm conditions in the untreated patients. Six patients had AML including acute promyelocytic leukemia, two had acute lymphoblastic leukemia (ALL), eight had nonHodgkin lymphoma (NHL), five had lymphoblastic lymphoma (LBL) and one had myelodysplastic syndrome. The median age of the untreated patients was 27.5 years. All except two patients showed normal sperm concentration and motility. One patient (case 2) had normal sperm concentration but deteriorated sperm motility, while another (case 15) had extreme oligospermia. Table 2 shows a summary of sperm conditions in the treated patients. The treated patients included seven patients with AML, three with ALL, one with chronic myelogeneous leukemia, two with NHL and three with LBL. The median age of the treated patients was 32 years. Semen was repeatedly A. Meguro K. Muroi (&) T. Matsuyama M. Mori T. Nagai K. Ozawa Division of Hematology, Department of Medicine, Jichi Medical University, 3311-1 Yakushiji, Shimotsuke, Tochigi 329-0498, Japan e-mail: muroi-kz@jichi.ac.jp

Sperm ultrastructure and meiotic segregation in a group of patients with chronic hepatitis B and C

Little is known about the effect of chronic hepatitis B and hepatitis C on sperm quality. In this study we analysed sperm quality from selected patients with chronic hepatitis B virus (HBV) or hepatitis C virus (HCV) infections. Semen samples were examined by light and transmission electron microscopy (TEM). TEM data were elaborated with a mathematical formula able to indicate a fertility index and the presence of the three main sperm pathologies: apoptosis, immaturity and necrosis. Meiotic chromosome segregation was investigated by fluorescence in situ hybridisation carried out on sperm nuclei, using probes for chromosomes 18, X and Y. Despite normal sperm concentration, we observed reduced motility. TEM analysis highlighted that 35.7% of patients showed generally good semen quality. However, significantly higher values of apoptosis and necrosis, compared with controls, were observed, demonstrating spermatogenetic alterations. Regarding meiotic segregation, we found an incidence of disomies similar to that observed in control samples, whereas diploidy resulted higher in HCV patients, without reaching statistical significance. In conclusion, sperm quality in the studied group was not impaired, however, apoptosis and necrosis resulted out of normal range and the fertility index was significantly lower in HCV and HBV infected patients versus controls.

Polymorphisms of the luteinizing hormone/chorionic gonadotropin receptor gene: association with maldescended testes and male infertility

Maldescended testes are the most common genital anomaly in newborns and are associated with testicular malignancy and infertility. As the inguinoscrotal phase of testis descent is androgen-dependent and requires integrity of the luteinizing hormone/chorionic gonadotropin receptor (LHCGR), we investigated whether nonsynonymous polymorphisms of the LHCGR gene are associated with maldescended testes. This was a retrospective case-control study including 278 patients with maldescended testes, 277 infertile men without maldescensus and 271 controls with normal sperm concentrations. Clinical and endocrinological workup of the patients was performed. Single nucleotide polymorphism (SNP) analysis was performed by GeneScan and TaqMan technology. Men with maldescended testes had significantly lower testis volumes, higher serum luteinizing hormone (LH) and follicle-stimulating hormone (FSH) but similar testosterone levels compared with both the control groups. The insLQ polymorphism in exon 1 (rs4539842) and the N291S SNP in exon 10 (rs12470652), showing increased receptor sensitivity in vitro, were not differently distributed between patients and controls. The S312N SNP in exon 10 (rs2293275) was significantly less frequent in men with maldescended testes than in controls. This difference was confirmed when infertile men with and without maldescensus were considered together. In men with maldescensus, a high LH drive maintains normal testosterone levels but this LH resistance is not associated with any particular LHCGR genotype. A significant association with the S312N polymorphism in exon 10 of the LHCGR is correlated to the spermatogenetic damage rather than to the maldescensus itself. Either the LHCGR itself or another genomic region linked to this SNP, possibly the germ cell-specific TFIIA-alpha/beta-like factor gene transcribed from the same genomic region in the opposite direction, is a risk factor for male infertility.

Morphology and Meiotic Segregation in Spermatozoa From Men of Proven Fertility

Estimates of semen parameters are important for defining normal ranges, which are currently established by 1999 World Health Organization guidelines. However, it is well known that semen evaluation is questionable because it is necessary for only 1 sperm to be able to reach and fertilize the oocyte. Spermiogram parameters and sperm morphology, evaluated by transmission electron microscopy (TEM), were performed on semen samples from 25 men of proven fertility. Despite a generally normal sperm concentration, progressive motility was reduced in 9 cases. Sperm characteristics were evaluated with an established technique, and the mean of the percentages of sperm pathologies were confirmed by comparing to previous reports. A comparison of apoptosis and necrosis in the samples, as detected by TEM and an annexin V/propidium iodide assay, was also performed. Fluorescence in situ hybridization was carried out on the same samples using probes for chromosomes 18, X, and Y. The mean value of the frequency of total aneuploidy in the analyzed group was 0.627% (25th percentile = 0.563%; median = 0.625%; 75th percentile = 0.690%). This study of the incidence of disomy and diploidy in spermatozoa from fertile, apparently normal individuals is important for making comparisons with infertile cohorts to determine the real increase of aneuploidy in those cohorts.

Genetic polymorphism of glutathione S-transferase T1 gene and susceptibility to idiopathic azoospermia or oligospermia in northwestern China

To investigate the association of glutathione S-transferase T1 (GSTT1) gene polymorphism in patients with idiopathic azoospermia or oligospermia in the northwestern China population. In the case-control study, GSTT1 genotypes were identified by multiplex polymerase chain reaction (PCR) with peripheral blood DNA samples from 78 patients with idiopathic azoospermia, 103 patients with idiopathic oligospermia and 156 age-matched controls with normal sperm concentration and motility, according to the criteria adapted from World Health Organization guidelines. All of the patients and controls were from northwestern China. There is a significant association between GSTT1 null genotype with idiopathic azoospermia risk (odds ratio [OR]: 2.36, 95% confidence interval [CI]: 1.33-4.20, P=0.003) or idiopathic oligospermia risk (OR: 2.00, 95% CI: 1.17-3.27, P=0.010). GSTT1 null genotype is a predisposing risk factor for sporadic idiopathic azoospermia or oligospermia in northwestern China.

Male Age and Sperm Necrosis in Assisted Reproductive Technologies

Introduction: Sperm apoptosis is well characterized but studies on the effect of male age and necrozoospermia are lacking. The objectives were: (a) to analyze percentages of apoptotic and necrotic sperm in ejaculates, and (b) to compare the results between younger and older age groups. Materials and Methods: Routine semen analyses were carried out (n = 189 males) and sperm cells were analyzed by dual fluorescence assay Hoechst 33342 and propidium iodide, and the acridine orange test. Results: The percentage of necrotic sperm in the ejaculate increased by 22% for males aged over 35. There was a positive correlation between age and necrosis (R = 0.30). Sperm apoptosis increased by 17% in males aged 45 and older. The population of DNA intact sperm declined in males aged 40 and over (R = –0.21). There were no age-related changes in strict normal morphology, sperm concentration and semen volume. A decrease in rapid progressive motility was correlated (R = –0.24) with male age and was significant after age 35. Conclusions: The study demonstrated increased necrosis, DNA damage and apoptosis while rapid progression and total motility declined with advancing age in the male beginning as early as age 35. The order of the observed changes was sequential, suggesting the involvement of different pathways in sperm necrosis after age 40.

Neutralization of Cytokine Activity at the Receptor Level Improves Sperm Motility in Men With Spinal Cord Injuries

Men with spinal cord injury (SCI) have a unique semen profile characterized by normal sperm concentrations but abnormally low sperm motility. Previous studies showed that elevated concentrations of cytokines in the seminal plasma of these men contribute to this condition. For example, when elevated concentrations of interlekin-1beta (IL-1beta), IL-6, and tumor necrosis factor-alpha (TNF-alpha) were immunoneutralized in the semen of men with SCI, sperm motility improved. The present study investigated if these cytokines act on sperm cell receptors to inhibit sperm motility. Semen was collected from men with SCI and from healthy non-SCI men. Sperm were separated from the seminal plasma by centrifugation. Eight identical aliquots of 5,000 sperm suspended in 50 microL of seminal plasma were prepared for each subject. Agents were added to the aliquots in order to neutralize IL-1beta, IL-6, and TNF-alpha at the receptor level. In SCI subjects, sperm motility improved in each treatment group compared with the untreated group, but statistical significance was reached only when neutralizing agents to all 3 cytokines were added. Improvement was less pronounced in subjects with close to normal semen cytokine concentrations or close to normal pretreatment sperm motility. In control subjects, IL-1beta, IL-6, and TNF-alpha were within normal values, and addition of receptor blockers to semen had no effect on sperm motility. These data support the hypothesis that cytokines act at the level of the sperm receptor to inhibit sperm motility. These data further support the notion that inactivating semen cytokines leads to improved sperm motility in SCI men. Our goal is to develop this finding into a treatment for low sperm motility in men with SCI.

Seminal tumor necrosis factor-alpha level and sperm nuclear DNA integrity in normozoospermic men

DNA damage in the male germ line is associated with poor semen quality, low fertilization rates or impaired preimplantation development. The causes of this DNA damage are still uncertain but the major candidates are oxidative stress and aberrant apoptosis. Tumor necrosis factor-alpha (TNF-a), a proinflammatory cytokine, is able to induce apoptosis and act as a proapoptotic factor. In vitro study demonstrated that co-incubation of TNF-a with sperms decreases total motility and produces chromatin and DNA fragmentation in a concentration- and time-dependent manner. To date, seminal concentration of TNF-a in relevant to sperm nuclear DNA integrity has not been studied. The present study aimed to evaluate seminal concentration of TNF-a in correlation with sperm parameters and nuclear DNA integrity in normozoospermic men. Prospective study. Forty-five ejaculated semen samples were obtained from healthy donors (n = 41) or male partners undergoing infertility investigation (n = 4) with age ranged from 25 to 50 years. All of subjects had no signs of genital inflammation and had normal sperm concentration (mean ± SD, 648.2 ± 273.5×106/mL). Sperm motilities were ranged between 22.7% and 86.3%. Routine sperm quality was assessed by computer-assisted semen analysis (CASA) and nuclear DNA integrity was measured by the TdT (terminal deoxynucleotidyl transferase)-mediated dUTP nick-end labelling (TUNEL) assay in raw semen. After centrifugation without washing, seminal plasmas were separated and then immediately frozen. TNF-a concentrations were measured by enzyme-linked immunosorbent assay (ELISA) in thawed seminal plasmas. Statistical analysis was performed using MedCalc. Sperm DNA fragmentation rates were ranged between 1.9% and 53.0% (mean ± SD, 12.4 ± 9.6%). Univariate analysis revealed that DNA fragmentation rate was not associated with sperm concentration and motility but had a negative correlation with linearity (r = −0.3247, P=0.0295). Seminal TNF-a was not detectable in 12 subjects (<1.0 pg/mL). The mean seminal concentration of TNF-a was 4.9 pg/mL with a range from 1.1 to 22.6 pg/mL. The TNF-a concentration had no correlation with clinically relevant parameters of sperm quality or nuclear DNA fragmentation rate. Our results indicate that sperm nuclear DNA fragmentation is not associated with seminal TNF-a level or sperm quality in asymptomatic normozoospermic men. Sperm nuclear DNA fragmentation in asymptomatic normozoospermic men may be not caused by seminal TNF-a.

Retained Functional Integrity of Bull Spermatozoa after Double Freezing and Thawing Using PureSperm® Density Gradient Centrifugation

The main aim of this study was to compare the motility and functional integrity of bull spermatozoa after single and double freezing and thawing. The viability and morphological integrity of spermatozoa selected by PureSperm density gradient centrifugation after cryopreservation of bovine semen in two commercial extenders (Experiment 1) and the function of bull spermatozoa before and after a second freezing and thawing assisted by PureSperm selection (Experiment 2) were examined. On average, 35.8 +/- 12.1% of sperm loaded onto the PureSperm density gradient were recovered after centrifugation. In Experiment 1, post-thaw motility and acrosome integrity were higher for spermatozoa frozen in Tris-egg yolk extender than in AndroMed, whether the assessments were made immediately after thawing [80.4 +/- 12.7 vs 47.6 +/- 19.0% motile and 78.8 +/- 8.3 vs 50.1 +/- 19.5% normal apical ridge (NAR), p < 0.05] or after preparation on the gradient (83.3 +/- 8.6 vs 69.4 +/- 15.9% motile and 89.5 +/- 7.2 vs 69.1 +/- 11.4% NAR, p < 0.05). For semen frozen in Tris-egg yolk extender, selection on the PureSperm gradient did not influence total motility but significantly improved the proportion of acrosome-intact spermatozoa. After the gradient, both the total motility and percentage of normal acrosomes increased for spermatozoa frozen in AndroMed (Minitüb Tiefenbach, Germany). In Experiment 2, there was no difference in sperm motility after the first and second freeze-thawing (82.9 +/- 12.7 vs 68.8 +/- 18.7%). However, the proportion of acrosome-intact spermatozoa was significantly improved by selection through the PureSperm gradient, whether measured by phase contrast microscopy (78.9 +/- 9.7 vs 90.4 +/- 4.0% NAR, p < 0.05) or flow cytometry (53.4 +/- 11.7 vs 76.3 +/- 6.0% viable acrosome-intact spermatozoa, p < 0.001). The improvement in the percentage of spermatozoa with normal acrosomes was maintained after resuspension in the cooling extender and cooling to 4 degrees C (88.2 +/- 6.2) and after re-freezing and thawing (83.6 +/- 6.56% NAR). However, flow cytometric assessment of the sperm membranes revealed a decline in the percentage of viable spermatozoa with intact membranes after the second freezing and thawing compared with after gradient centrifugation (76.3 +/- 6.0% vs 46.6 +/- 6.6%, p < 0.001) to levels equivalent to those obtained after the first round of freeze-thawing (53.4 +/- 11.7% viable acrosome-intact spermatozoa). Sperm movement characteristics assessed by computer-assisted analysis were unaffected in the population selected on the PureSperm gradients but declined after cooling of the selected and extended spermatozoa to 4 degrees C. There was no further change in these kinematic measurements after the cooled spermatozoa had undergone the second round of freeze-thawing. These results demonstrate that bull semen can be frozen and thawed, followed by a second freeze-thawing cycle of a population of spermatozoa selected by PureSperm, with retained motility and functional integrity. This points to the possibility of using double frozen spermatozoa in bovine artificial insemination programmes and to the potential benefits of PureSperm density gradient centrifugation for the application of cryopreserved bull spermatozoa to other biotechnological procedures such as flow cytometric sex sorting followed by re-freezing and thawing.

Sperm count and sperm motility in incidental high-grade varicocele

One hundred twenty-four otherwise healthy men with suspected high-grade varicocele, comprised of 28 patients with grade 2 and 96 with grade 3 varicocele, were referred to us by their primary care providers to determine their exact grade so that they could be exempted from the compulsory armed force service if they had grade 3 varicocele. In the grade 2 and grade 3 groups, mean sperm concentration and mean sperm motility were 80.89 million per milliliter (SD, 38.98 million/mL; median, 74.5 million/mL; range, 22-210 million/mL) and 78.77 million per milliliter (SD, 41.34 million/mL; median, 77 million/mL; range, 0-223 million/mL) and 58.39% (SD, 13.7%; median, 60%; range, 22-75%) and 60.31% (SD, 16.93%; median 65%; range, 0-80%), respectively, with no statistically significant differences between the two groups, no patient with infertility as a complaint during a 20-month period, and no patient with sperm concentrations of <20 million/mL, but with six patients with sperm motilities <50%. In summary, by far the great majority of patients with high grade varicocele have more than normal sperm output. Higher grades of varicocele are not associated with more pronounced deleterious effects on sperm concentration and percentage motility. Incidentally diagnosed high-grade varicocele is not a progressive condition, and almost all patients retain normal semen quality, at least over a period of 20 months.

Relationship between age and semen parameters in men with normal sperm concentration: analysis of 6022 semen samples

This study evaluates retrospectively the relationship between age and semen parameters among men with normal sperm concentration. It was based on computerized data and performed in an Academic Fertility and IVF Unit. Six thousand and twenty-two semen samples with sperm concentrations of >or=20 x 10(6) ml(-1) were examined according to WHO criteria and analysed in relation to patients' age. For each age group, mean values +/- SD of semen volume, sperm concentration, percentage of motile spermatozoa, normal morphology, acrosome index, total sperm count/ejaculate, total motile sperm count/ejaculate and sexual abstinence duration were examined. A peak semen volume of 3.51 +/- 1.76 ml(-1) was observed at age >or=30 to <35 years and a lowest volume of 2.21 +/- 1.23 ml(-1) was observed at age >or=55 years (P<0.05). Sperm motility was found to be inversely related to age with peak motility of 44.39 +/- 20.69% at age <25 years and lowest motility of 24.76 +/- 18.27% at age >or=55 years (P<0.05). A reduction of 54% was observed for total motile sperm, between values of 103.34 +/- 107 x 10(6) at age >or=30 to <35 years and 46.68 +/- 53.73 x 10(6) (P<0.05) at age >55 years. A statistically significant and inverse relationship was observed between semen volume, sperm quality and patient age, in spite of prolonged sexual abstinence duration. Top sperm parameters were observed at age >or=30 to <35 years, while the most significant reduction in sperm parameters occurred after the age of 55 years.

Spermatozoa and Chronic Treatment with Finasteride: A TEM and FISH Study

Finasteride is a specific inhibitor of the 5alpha reductase enzyme originally approved for the treatment of benign prostatic hypertrophy and also for the treatment of androgenetic alopecia (AGA) in men at a dose of 1 mg/day. We report on three cases of young men recruited at our Centre for Male Infertility who had used finasteride for five years. Semen quality was investigated by light microscopy to evaluate sperm concentration and motility. Sperm morphology was performed by transmission electron microscope (TEM) and the data were analyzed. The presence of Y microdeletions was investigated by PCR. Meiotic segregation was explored by fluorescence in situ hybridization (FISH). Patient 1 was azoospermic, patients 2 and 3 showed a normal sperm concentration and severely reduced progressive motility. TEM analysis revealed altered sperm morphology consistent with necrosis and FISH data revealed elevated diploidy and sex chromosome disomy frequencies. This examination was repeated 1 year after the men had suspended the use of finasteride, without receiving any other treatment. A recovery of spermatogenetic process was observed. Motility and morphology improved whereas the meiotic pattern did not change presenting elevated diploidy and sex chromosome disomy frequency.

A análise seminal deve ser requisitada para homens com histórico de fertilidade prévia?

PURPOSE: to determine if the previous fertility history can predict current fertility status of a patient examined for couple’s infertility. METHODS: retrospective study involving semen analyses from 183 consecutive subfertile patients evaluated from September 2002 to March 2004. We excluded those patients who had undergone radio or chemotherapy, orchiectomy or vasectomy. Mean values of all analyses were used for patients with multiple semen analysis. Patients with more than 20x106 sperm/mL, motility higher than 50% and with normal strict sperm morphology higher than 14% were considered normal. Patients were divided into two groups, according to the fertility status: primary infertility (118 patients) and secondary infertility (65 patients). Data were analyzed according to the c2 test and the Student t-test. RESULTS: no differences were detected in the mean age between patients with primary infertility, 37.3±6.3, and secondary infertility, 38.1±5.9; p=0.08. In the group of patients with primary infertility, 51.9% (61 patients) had a normal sperm concentration, 70.3% (83 patients) had normal sperm motility and 26.3% (31 patients) had normal sperm morphology. In the group of patients with secondary infertility, 53.8% (35 patients) had normal sperm concentration, 75.4% (49 patients) had normal sperm motility and 32.3% (21 patients) had normal sperm morphology. No significant differences were detected in sperm concentration (21.3x106/mL versus 23.1x106/mL; p=0.07), motility (45.2 versus 48.1%; p=0.08) and morphology (6.1 versus 6.4%; p=0.09) between groups of patients with primary and secondary infertility. CONCLUSIONS: semen analysis should be requested even in cases of prior male fertility. Physicians should not presume a patient to have a normal semen analysis based on his previous history of initiating a pregnancy.

Expression of the HSPA2 gene in ejaculated spermatozoa from adolescents with and without varicocele

To evaluate mRNA expression of the HSPA2 gene in ejaculated spermatozoa in adolescents with and without varicocele. Controlled prospective study. Patients in an academic research environment. Adolescent patients with clinical diagnosed bilateral varicocele grades II and III, and adolescent patients without varicocele. Reverse-transcription-polymerase chain reaction (RT-PCR) analysis of HSPA2 gene expression in adolescents with and without varicocele. Comparative RT-PCR expression analysis of HSPA2 gene mRNA, compared to the housekeeping beta-actin gene. Sperm from adolescents with varicocele and oligozoospermia had significantly lower levels of HSPA2 gene expression than both adolescents without varicocele (controls) and adolescents with varicocele and normal sperm concentration (P<.05). The latter group had a nonsignificant increase in gene expression compared to the control group. This is the first report on HSPA2 gene expression in ejaculated spermatozoa from adolescents and its relationship with varicocele pathology. Results demonstrated that HSPA2 expression was down-regulated in adolescents with varicocele and oligozoospermia compared to controls. There was a higher, albeit nonsignificant, gene expression in adolescents with varicocele and normal sperm concentration than in controls. We speculate that expression levels of this gene might be used as a molecular marker for the acquisition of thermal tolerance in ejaculated spermatozoa.