Showing papers in "Archives of Andrology in 2000"

Psychosocial response of Chinese infertile husbands and wives.

Abstract: Some 15% or 1 in 6 American couples in the childbearing years have infertility problems. Numerous studies have demonstrated that both infertile men and women have negative emotional responses, such as stress, anxiety and depression. In Europe, Canada, and the United States the response of infertile husbands was different from that of their wives in self-image, marital adjustment, and sexual relations. The differences in psychological distress, marital satisfaction, and sexual satisfaction between Chinese infertile husbands and wives were evaluated. Fifty-nine infertile couples participated in this study. The subjects completed an Infertility Questionnaire, Marital Satisfaction Questionnaire, and Sexual Satisfaction Questionnaire as measures of gender differences in facing infertility problems. Paired t tests revealed that husbands expressed significantly less distress than that of the wives. The husbands' self-esteem was higher than that of the wives. The husbands' marital and sexual satisfaction was also higher than that of the wives. These results propose that although differences exist in cultural, ethnic, and religious norms between Chinese society and Western society, the Chinese couples' response to infertility is similar to that of Western couples. The major difference is that the in-laws play an important role in Chinese society, especially in marital satisfaction.

Participation of DNA structure on sperm chromatin organization.

Abstract: The in vitro interaction between purified bovine liver and sperm DNA with somatic histones, to form nucleosomes, and with bovine and salmon protamines were studied. DNAse or microccocal nuclease digestion of liver DNA-histone reassociated chromatin produced the expected polynucleosome type of fragments. Electrophoretic patterns of digested sperm-DNA nucleosomes were different. Micrococcal nuclease digestion produced mainly fragments smaller than 100 bp and some nucleosome-type particles. Under DNAse activity most of the products were smaller than 100 bp, indicating an increased susceptibility of the sperm DNA-histone complexes to the hydrolytic activity of both nucleases, particularly toward DNAse I. This differential susceptibility was confirmed by sucrose gradient spectrophotometric analysis. Acridine orange (AO) staining of histone-DNA reassociated nucleosomes showed significant differences in fluorescence intensity, sperm DNA-histone complexes being almost twice as fluorescent as liver DNA-histone complexes. On the contrary, liver DNA/protamine complexes stained with AO were consistently more fluorescent than sperm DNA-protamine complexes. Finally, no differences in either fluorescence intensity or spectra were observed when liver and sperm DNA were stained with AO after interaction with salmon protamines. The data suggest that sperm DNA has important structural characteristics that differentiates it from somatic DNA. These differences seem to be species specific and must surely play an important role on the determination of the dramatic sequence of that participates sperm chromatin organization.

Poor quality of sperm as it affects repeated early pregnancy loss.

Abstract: A study was carried out to determine whether males contribute to repeated early pregnancy loss. Semen samples were analyzed from proven-fertile men (n = 51 group I) and from men whose partners presented with early pregnancy loss (>3 first trimester abortions, n = 32 group II). Routine analysis, sperm function tests, and ultrastructural studies of sperms were carried out. Female factor could be identified in 25 (78%) couples, and in 7 (22%) no cause either male or female could be identified and the semen analysis was normal. Percent morphologically normal did not differ significantly between the groups, but increased sperm head abnormalities were seen. The functional tests were all normal except for a significant decrease in the capacity of nuclear chromatin to decondense in vitro. The ultrastructural studies showed defects of chromatin condensation and irregular nuclei with vacuoles. This study points to the loss of chromatin integrity as a possible contributing factor from males to early pregnancy loss.

Antioxidative effect of melatonin on human spermatozoa.

Abstract: The ability of melatonin to suppress experimentally induced lipid peroxidation (LPO) in sperm membrane was investigated in 41 samples of infertile men. Iron/ascorbate (0.04/0.2 mmol)-induced LPO was measured by the formation of malondialdehyde (MDA) using the thiobarbituric acid method. Sperm incubated in the presence of melatonin (2-6 mmol) exhibited a concentration-dependent decrease of MDA generated from hydroperoxide of the sperm plasma membrane in the presence of promoter system. Addition of 6 mmol of melatonin significantly reduced the rate of lipid peroxidation in sperm of unselected donors (mean +/- SE in control samples = 26.4 +/- 2.9 vs. 6.5 +/- 1.1 nmol MDA/108 sperm in melatonin-treated samples; n = 16, p <. 005). Inhibitory effect of melatonin was also significant in the presence of 0.015 mmol of ferrous ions (20.5 +/- 1.7 vs. 7.9 +/- 1.6 nmol MDA/108 sperm in melatonintreated samples; n = 7, p <. 02) and 0.005 mmol of ferrous ions (20.2 +/- 2.8 vs. 9.9 +/- 2.4 nmol MDA/ 108 sperm; n = ...

Nonsperm cells in human semen and their relationship with semen parameters.

Abstract: The prevalence and clinical significance of leukocytes (WBC) and immature germ cells in semen is currently a matter of controversy. The aim of this work was to assess the prevalence of leukocytospermia in semen samples from Venezuelan men and its possible effects on sperm parameters. The concentration of WBC and round cells (RC) was evaluated in 118 semen samples from 19 fertile subjects (group 1), 62 infertile patients (group II), and 37 men with varicocele (group III). Semen WBC concentration was assessed by peroxidase assay. Twenty-six (22%) of the total samples had more than 10 WBC/mL semen. Twenty of the infertile men had leukocytospermia (32%) compared with 16% in the fertile group and 8% in the varicocele group. Semen RC concentration was lower than 5 x 10(6)/mL in all groups but, in groups II and III was significantly higher compared with group I. Infertile men had the highest WBC concentration. WBC concentration was negatively correlated with progressive motility, percentage of morphologically normal sperm, and hypoosmotic swelling test in infertile men but not in the varicocele group. In this group a negative correlation was obtained between immature germ cells and normal sperm morphology. The data show that leukcytospermia occurs frequently in infertile patients and is associated with poor semen quality parameters. In contrast, in men with varicocele, the increased number of immature germ cells might play a pivotal role in the pathogenesis of abnormal spermatozoa.

Role of estrogens in human benign prostatic hyperplasia.

Abstract: The aging process is associated with a progressive decline of plasma testosterone levels, while estrone and estradiol remain unchanged and sex hormone binding globulin (SHBG) increases, with reduction of bioavailable testosterone in prostatic tissue with benign prostatic hyperplasia (BPH) the most important androgen is dihydrotestosterone: with its receptors it is almost uniformly distributed in the epithelial and stromal compartment and is not supranormal. Intraprostatic estrogens and their receptors are elevated and concentrated in the stroma. Androgens may act on the prostate indirectly through the production of growth factors; in human BPH no clear evidence exists on the modulatory effect of estrogens on bFGF, KGF and TGFbeta formation. A western diet, characterized by high fat consumption, predisposes men to BPH, while a diet rich in flavonoids and lignanes, containing phyto-estrogens, lowers this risk. These data suggest that in the medical treatment of BPH, antiestrogens or aromatase inhibitors may be used: however, up to now the clinical results of this treatment are not promising and the improvement of the obstructive symptoms does not exceed that of placebo. A possible explanation of this unsatisfactory result could be that the estrogen reduction secondary to the use of aromatase inhibitors is counterbalanced by the rise of androgen precursors.

ICSI as an effective therapy for male factor with antisperm antibodies

Abstract: This study was conducted to evaluate if in vitro fertilization (IVF) with intracytoplasmic sperm injection (ICSI) is an effective treatment for infertility complicated by the presence in the male partner of sperm autoantibodies. Over a 1-year study period comparisons of fertilization, pregnancy, and implantation rates were made in couples where the male partner was negative or weakly positive for sperm autoantibodies ( 80%) (gr 2); or autoantibodies were moderately positive (50-80%) (gr 3). Only patients having oocytes fertilized by ICSI were included. The fertilization, clinical pregnancy, implantation, and miscarriage rate for group 1 (n = 67) was 56, 43, 21, and 14%. Comparable values for group 2 (n = 20) were 55, 40, 23, and 25%, and for group 3 (n = 6) were 63, 33, 23, and 0%. IVF with ICSI demonstrates comparable fertilization, pregnancy, implantation, and miscarriage rates in female partners of males with and without sperm autoantibodies.

Mechanism of ejection during ejaculation: identification of a urethrocavernosus reflex.

Abstract: The ejaculatory mechanism involves 2 reflexes: the "glans-vasal", which seems to bring the semen to the posterior urethra (emission phase of ejaculation), and the "urethromuscular" which ejects it to the exterior (ejection phase). This study investigated the mechanism of bulbocavernosus muscle (BCM) contraction, once the seminal fluid reaches the bulbous urethra. The study included 14 healthy male volunteers (mean age 37 +/- 10.2 SD years). To test the response of the BCM to urethral distension, a 10F balloon-tipped catheter was introduced into the prostatic urethra and filled with saline in increments of 0.25 mL; a needle electrode recorded the response. The balloon was then withdrawn to lie in the membranous, bulbous, and pendulous urethra and the test was repeated at each site. The latency of the muscle response was calculated. The BCM response to each of the anesthetized bulbous urethra and anesthetized BCM was recorded. Distension of the prostatic, membraneous, or pendulous urethra effected no BCM EM...

Schedule to inject in vitro matured oocytes may increase pregnancy after intracytoplasmic sperm injection

Abstract: To ascertain the value of using immature oocytes in an intracytoplasmic sperm injection (ICSI) program, the authors designed a schedule, at 5 p.m. on day 1 (the day of oocyte retrieval) and at 8 a.m. and 2 p.m. on day 2, to recognize and inject the in vitro matured (IVM) oocytes. For the 1,166 oocytes retrieved in 107 ICSI cycles, 128 (11.0%) were at the stage of metaphase I (MI) and 113 (9.7%) at germinal vesicle. Routine ICSI for metaphase 11 oocytes was performed at 2 p.m. on day 1 (initial ICSI). In culture medium of human tubal fluid with 15% maternal serum, 85.1% (205/241) immature oocytes progressed to maturation in which 16.4% (21/128) of MI oocytes matured at 5 p.m. of day 1. The rate of normal fertilization for IVM oocytes (58.5%) was not significantly different from that of initial ICSI (64.0%). One patient received a transfer of two fertilized IVM oocytes alone that were injected at 5 p.m. of day 1, maturing from the MI stage, and achieved a normal pregnancy. The fertilized IVM oocytes were replaced along with the embryos from initial ICSI for 40 cycles that led to 14 (35%) clinical pregnancies. In 43 fertilized IVM oocytes donated for research, we observed that cleavage (95.3%) to the 2- to 4-cell stage was not distinct from that of initial ICSI (94.6%); however, the percentage of embryos of grade I and II morphology was significantly smaller (24.4% vs. 62.5%). Only five (11.6%) developed to blastocysts in vitro. Twenty-one fertilized IVM oocytes were frozen for future transfer. A schedule to inject IVM oocytes in ICSI cycles may generate more accessible embryos for fresh transfer or cryopreservation to increase the chance of pregnancy, although the embryo quality was relatively poor.

Ultrastructural interrelationship between the pineal gland and the testis in the male rat

Abstract: The ultrastructural interrelationship between the pineal gland and testis was evaluated in the rat. Wistar rats were divided into 6 groups. Groups I and II were sham-orchidectomized and orchidectomized rats, respectively. Rats in group III were orchidectomized and daily injected with testosterone propionate (TP) for 1 month. Groups IV and V were sham-pinealectomized and pinealectomized, respectively. Group VI was pinealectomized and daily injected with melatonin for 2 months. All animals were anesthetized with ketamine for fixation by vascular perfusion. Pineal glands of groups I, II, and III and the testes of groups IV, V, and VI were removed and weighed. All specimens were examined by electron microscopy. Orchidectomy caused an increase of lipid droplets, cytoplasmic dense bodies, and lysosomes. Rough endoplasmic reticulum, Golgi apparatus, and mitochondria were extensive in the cytoplasm. TP administration to orchidectomized rats resulted in formation of less extensive lipid droplets and mitochondria. In pinealectomized rats, golgi complex, mitochondria, and enlarged smooth endoplasmic reticulum were extensive in the cytoplasm of Leydig cells. Formation of cytoplasmic secretory granules and osmiophilic bodies was observed. Testicular weight increased compared to group IV. Melatonin decreased testicular weight in comparison to group V and prevented ultrastructural changes. Pinealectomy and orchidectomy caused hyperactivity in Leydig cells and pinealocytes, respectively, which suggests a mutual relationship between the pineal gland and testis in the rat.

Semen analysis at the turn of the century: an evaluation of potential uses of new sperm function assays

Abstract: Semen analysis is a critical assay in the evaluation of infertility and may yield critical information regarding the etiology and prognosis of many types of reduced male fertility. However, basic semen analysis does not directly measure sperm fertilizing capacity, or many of the biochemical events both prior to and subsequent to fertilization. In the last two decades numerous assays of sperm function have been developed. These assays can be classified as: 1) Assays of general biochemistry and ultrastructure, 2) Assays of zona binding and oocyte penetration, and 3) Assays of postpenetration events. Sperm function assays not only allow an accurate diagnosis of many infertilities not diagnosed by the semen analysis, but can also lead to improved treatment modalities. In this review, basic semen analysis and many sperm function assays are briefly reviewed. Novel uses of sperm function are demonstrated in brief case studies.

Liquid storage (5°c) of ram semen in different diluents

Abstract: This study examined the effects of three diluents - skim milk (M), TesT (TE) and Tris-trehalose (TR) - on quality of ram semen stored at 50C over a long term. Percentage of motile spermatozoa (PM), motility score (MS), percentage of intact acrosomes (PIA), and percentage of swollen spermatozoa under hypoosmotic conditions (PS) were assessed after cooling and every 24 h up to day 16. There was a significant effect (p 50% up to day 8, while TE and M were greater than 50% up to day 5. Skim milk maintained progressive movement of sperm cells for longer (MS=3 until day 5) than the other diluents. There were no significant differences in...

Role of prosaposin in the male reproductive system: effect of prosaposin inactivation on the testis, epididymis, prostate, and seminal vesicles

Abstract: SGP-1/prosaposin can be secreted or targeted to the lysosomes where it is processed into smaller saposins (A, B, C, and D) required for the hydrolysis of glycosphingolipids. The deficiency of saposins B and C results in variant forms of metachromatic leukodystrophy and Gaucher's disease, respectively, which are characterized by lysosomal storage of undegraded glycosphingolipids. In the nervous system, prosaposin presents trophic activity. A mouse model was recently developed by creating a null allele in embryonic stem cells through gene targeting to investigate the phenotypic diversity of prosaposin mutations and the involvement of this protein in lysosomal storage diseases, and for the development of therapeutic approaches. Mice homozygous mutants die at the age of 35-40 days and neurological disorders contribute to the early demise of the mutant mice. The male reproductive organs in homozygous mutants show several abnormalities, such as a decrease in testis size with reduced spermiogenesis and an involution of the prostate, seminal vesicles, and epididymis. In these animals, the blood levels of testosterone remain normal. In the prostate of homozygous mutants, only the basal epithelial cells appear to be present, while the secretory cells are absent. These findings suggest that prosaposin may be involved in the development and maintenance of the male reproductive organs, as well as, in cellular differentiation.

Ultrastructural investigation of human sperm using atomic force microscopy.

Abstract: Ultrastructural investigation of human sperm in its natural environment (without fixation, dehydration, embedding, sectioning, etc.) was carried out by using atomic force microscope (AFM) in its tapping mode. This technique permits the examination of fine structural details of undamaged sperm and its topography with precision. Moreover, it allows 3D reconstruction of images and enhances the contrast to resolve details such as mitochondria that surround the axoneme at the sperm middle piece. An organized structure has been found in the flageller axoneme region. Ultrastructure also reveals folding and details of the depression of the membrane that cannot be examined with conventional techniques.

Low plasma testosterone in varicocele patients with impotence and male infertility.

Abstract: To study the affect of bilateral varicocele (grade 3) on impotence and male infertility patients, 29 patients were selected from an outpatient clinic during 15 May 1998 to 15 August 1999 (the mean age was 33.9 +/- 6.3), 15 patients complaining of erectile dysfunction and 14 patients complaining of male infertility. The mean duration of impotence was 3 +/- 2.3 years and for male infertility was 6 +/- 2.5. All organic and psychogenic causes related to impotence and male infertility except bilateral varicocele (grade 3) and low plasma testosterone were excluded by clinical and laboratory investigations. Twenty males with normal erection and fertility were included as controls. Detailed medical history and complete physical examination included measurement of testicular size by orchiometer; semen and hormonal parameters were measured for all patients and control. In impotent patients left and right testicular volume was significantly decreased (p < .05), while in male infertility patients left and right testicular volume was highly significantly and, significantly decreased (p < .005, p < .05) compared to controls. In male infertility patients, left testicular volume was highly significantly decreased compared to impotent patients (p < .005). The sperm count and semen volume in impotent patients was significantly decreased (p < .05, p < .01), but no significant differences were found in sperm motility and abnormal forms, while in male infertility the sperm count was highly significantly decreased (p < .005), the sperm motility was significantly decreased (p < .05), the abnormal form was significantly increased (p) < .05), but in the semen volume there was no significant difference compared to controls. In impotent patients the sperm count was significantly increased and abnormal form was significantly decreased compared to male infertility (p < .05). The mean serum testosterone was significantly decreased in impotent patients (p < .01), and highly significantly decreased in male infertility (p < .005) compared to controls. The mean serum FSH was significantly increased in male infertility (p < .05) and nonsignificant in impotent patients compared to controls. The mean serum LH and prolactin levels were nonsignificant in both impotent and male infertility patients compared to controls, but LH was significantly increased in impotence compared to male infertility patients (p < .025). Therefore, bilateral varicocele (grade 3) is associated with significant reduction in testicular function with significant increase in serum levels of FSH and LH, which may cause erectile dysfunction and male infertility.

Stimulation and scavestrogen-induced inhibition of reactive oxygen species generated by rat sertoli cells.

Abstract: The ability of Sertoli cells harvested from 18-day-old Sprague-Dawley rats to generate reactive oxygen species (ROS) occurs under unstimulated and stimulated conditions. Thus, the generation of ROS and its regulation by stimulating and inhibiting compounds was determined as a lucigenin-dependent chemiluminescence reaction. According to the data, ROS generation was influenced by different cell preparation conditions--stimulating substances such as PMA, FMLP, C5a, A23187, and scavestrogens characterized by antioxidant, radical-scavenging properties. The mechanical homogenization during cell preparation procedures leads to an increase of ROS generation. ROS generation of Sertoli cells was followed by elected substances in the following rank order of efficacy: PMA > FMLP > or = C5a > Ca-ionophore A23187. The registered inhibiting effects of the scavestrogens J811 and J861 were significant. The measured CL counts decreased at 72 and 77%, respectively, of control experiments done without scavestrogens. The generation of reactive oxygen species in Sertoli cells and especially the increase in oxygen free radicals and their effects on cellular structures of spermatids are directly involved in inducing morphological alterations. Sertoli cells play an important role in spermatogenesis. The measurements of ROS may have clinical relevance in the evaluation of infertility.

The use of ebastine, a mast cell blocker, for treatment of oligozoospermia

Abstract: The effect of mast cell blocker (ebastine) on semen quality was evaluated in 15 idiopathic oligozoospermic males. Ten milligrams/day of ebastine was administered orally for 3 months. Nine cases (66.7%) showed definite improvement in the semen quality. The wives of 3 patients (20%) became pregnant within 6 months of the initial treatment. It would appear that mast cell blocker (ebastine) significantly improves semen quality in men with idiopathic oligozoospermia.

Does seminal fluid viscosity influence sperm chromatin integrity

Abstract: A retrospective study was undertaken to investigate whether viscosity alters sperm chromatin integrity. Semen samples were obtained from 269 men attending the infertility clinic. The viscosity was measured quantitatively by needle and syringe method and the viscosity ratio was calculated against distilled water. The chromatin integrity was evaluated by in vitro decondensation test using 1% SDS and 6 mM EDTA. According to the viscosity ratios the samples were divided into 2 groups: I, normal (ratio 9, n = 30) viscosity. Chromatin integrity was significantly lower in the group with higher viscosity. Significant decrease in sperm count and motility were seen in group II as compared to group I. Thus, hyperviscosity of seminal fluid alters the sperm chromatin integrity.

Potassium increases intracellular calcium simulating progesterone action in human sperm.

Abstract: Progesterone (P) and zona pellucida are known to induce acrosome reaction in human sperm by increasing cytosolic calcium. High concentrations of potassium ions (K+) improve the rate of acrosome reaction in human sperm in vitro. This article determined whether the effect of K+ on the acrosome in human sperm is mediated by increasing intracellular calcium ([Ca2+]i). The effect of K+ on [Ca2+]i was examined by using Fura 2 as the fluorescent indicator. The effect of K+ and P on [Ca2+]i in sperm and the involvement of ion channels was compared. Motile sperm were collected by the swim-up method from semen of healthy volunteers and capacitated overnight in BWW containing 0.5% BSA. Incubation of capacitated sperm with different concentrations of potassium chloride (1.25-20 mM) resulted in dose-dependent increase in [Ca2+]i similar to that observed with P. The increase in [Ca2+]i by K+ and P was blocked by the addition of EGTA, a Ca2+ chelator. K+-induced change in [Ca2+] was not altered by the addition of dihydr...

Evidence that sperm with low hypoosmotic swelling scores cause embryo implantation defects.

Abstract: A previous prospective study using matched samples found that sperm with low hypoosmotic swelling (HOS) scores had no adverse effect on fertilization rates but markedly reduced pregnancy and implantation rates. The present study attempted to corroborate or refute the aforementioned study by comparing pregnancy rates in donor-recipient pairs using shared oocytes where the sperm of one of the two males had low HOS scores. The results found no pregnancies from the sperm with low HOS scores versus a 41% live delivered rate for those with normal scores. However, fertilization rates were not affected. This retrospective study thus confirms that sperm with low HOS scores cause implantation defects of the embryos that are formed without affecting fertilization rate, embryo cleavage rate, or embryo quality.

Functional activity of mouse sperm was not affected by low doses of aspirin-like drugs

Abstract: To investigate some possible effects of low doses of nonsteroidal anti-inflammatory drugs upon functional activity of mouse sperm, the authors injected lysine acetyl salicylate (im 14.3mg/kg day-1), ibuprofen (ip 5.6 mg/kg day-1), or piroxicam (ip 0.28 mg/kg day-1) to pregnant females (the male cohort was sacrificed at adulthood) (A) or to adult males during 35 (B) or 60 (C) days. Parameters measured were motility, viability, acrosomal integrity, responses to hypoosmotic shock, in vitro fertilization index, and testosterone plasma levels. Salicylate evoked a slight reduction in the percentage of swollen gametes in A, and ibuprofen diminished testosterone plasma levels in B. The other parameters remained unchanged in all groups. Results are well supported by the low doses assayed, which are equivalent to the content of one tablet commercially available for each compound.

Development of the coping scale for infertile couples

Abstract: Infertility treatment involves more complicated, uncomfortable, and humbling medical procedures for women, so the coping strategies used by wives may differ from those used by husbands. The purpose of this study was to develop and test a Coping Scale for Infertile Couples that would be sensitive to the differences in gender. What couples actually do to cope with infertility and whether these strategies are different between couples were also addressed. A total of 138 infertile couples participated in this study. The Coping Scale for Infertile Couples was administered with the Infertility Questionnaire, the Perceived Stress Scale, and the Jalowiec Coping Scale as measures of concurrent validity. Factor analysis extracted 4 subscales from the Coping Scale for Infertile Couples. Test-retest and internal consistency reliability were high. Paired t tests revealed that wives used the "Increasing space and Sharing the burden" strategies to a greater degree than their husbands. Husbands used the "Being the best" strategy to a greater degree than the wives. A significant correlation with distress, stress, and coping measures provided evidence of concurrent validity. Preliminary results suggest that this measure has good reliability and validity, which can contribute toward the elucidation of coping strategies used by infertile couples and assist in planning effective interventions.

A randomized study comparing the effect of standard versus short incubation of sperm and oocyte on subsequent pregnancy and implantation rates following in vitro fertilization embryo transfer.

Abstract: Some data suggest that some sperm are associated with a toxic product that lowers embryo implantation rates when these sperm come in contact with the zona pellucida. The possibility exists that shortening the incubation time could improve pregnancy rates. A randomized prospective study was initiated to evaluate pregnancy and implantation rates with transfer of embryos formed by conventional overnight vs short (2-h) co-incubation of sperm and oocytes. The clinical and viable pregnancy rates were 41 and 29% for conventional co-incubation versus 30 and 30% for short co-incubation. Implantation rates were also similar (17.9 vs 17.8%). Short incubation does not seem to impact any advantage over conventional co-incubation techniques in couples undergoing in vitro fertilization where the male factor is normal.

Pericentric inversion of the y chromosome of infertile male

Abstract: The authors report a case with pericentric inversion of the Y chromosome associated with asthenonecrozoospermia. The conventional karyotype was 46, X, inv (Y) (p11q11). Polymerase chain reaction (PCR) analysis revealed the deletion of DYZ3, DYS139, and RBM1. Three-color fluorescent in situ hybridization (FISH) analysis of the sperm chromosomes showed normal ratio between X- and Y-bearing sperm. In this case, the frequencies of aneuploidy of the sperm are not significantly higher compared with those from the normal volunteers. Cytogenetic analysis is recommended when the patients with pericentric inversion of the Y chromosome are attending an infertility clinic.

Sperm morphology and chromatin condensation before and after semen processing.

Abstract: Semen analysis constitutes the most important investigation of male infertility. However, the true anomalies present in defective sperm cells have been only partially characterized. The integrity of the sperm chromatin may play the most important role, particularly in ICSI, where most of the natural selection mechanisms are bypassed. This study was carried out to characterize sperm morphology (strict criteria), to evaluate chromatin condensation and sperm count in native semen as well as after semen preparation by the swim-up technique, and to eventually evaluate any correlation between these parameters. Semen from 90 men was analyzed for the above parameters in both the fresh and processed semen. Whereas the sperm count decreased after sperm preparation by the swim-up technique in comparison to the value in the fresh semen (p<.001), there was an increase in the percentage of morphologically normal (p<.001) and chromatin-condensed sperm (p=.99). However, there was no correlation between sperm morpho...

Antibodies to human sperm ylp12 peptide that is involved in egg binding inhibit human sperm capacitation/acrosome reaction

Abstract: Recently, the authors reported a novel dodecamer peptide sequence, designated as YLP12, on human sperm, that is involved in binding to zona pellucida (ZP) of human oocyte [10]. This unique sequence is present on the acrosomal region of the human sperm cell and is expressed only in human testis/sperm. The aim of the present study was to examine whether YLP12 sequence is involved in capacitation/acrosome reaction. Swim-up sperm were capacitated with anti-YLP12 Fab' antibodies or control Fab's (40 and 85 µg/mL) and then the acrosome reaction was induced with calcium ionophore. An average of 64-73% sperm underwent acrosome reaction when they were capacitated in the presence of 40-85 µg/mL of bovine serum albumin or control Fab's.A significant (p <. 01 to <. 001) reduction (58-75%) in the percentage of acrosome-reacted sperm was observed when the sperm were capacitated in the presence of YLP12 Fab's. These data indicate that the YLP12 peptide sequence is involved in sperm capacitation/acrosome reaction, ...

Human leukocyte antigen II expression in sperm cells: comparison between fertile and infertile men.

Abstract: Human leukocyte antigens (HLA) class II transcripts in mature spermatozoa of healthy volunteers have recently been demonstrated using reverse transcription-polymerase chain reaction (RT-PCR). HLA II expression was investigated on ejaculated sperm cells in fertile and infertile men by RT-PCR and flow cytometry. Among 22 fertile and 20 infertile men, 18 were selected for the study because they showed no contamination with non-sperm cells. HLA II mRNA transcripts were expressed in all but 1 of 8 infertile subjects and in only 2 of 10 fertile ones. The cytofluorometric analysis on three RT-PCR positive samples confirmed the presence of class II antigens on cell surfaces. These data clearly confirm the presence of both HLA II mRNA and surface molecules on human sperm cells. In addition, an interesting nonrandom distribution of positivity among fertile and infertile samples regarding HLA II expression (p <. 025) suggests a possible correlation with infertility.

Does the hypoosmotic swelling test predict human sperm viability

Abstract: Human sperm viability is essential for successful fertilization. Eosin Y is the usually accepted method for sperm viability assessment, though the hypoosmotic swelling test has been proposed for the selection of viable spermatozoa in procedures such as intracytoplasmic sperm injection. The present study was designed to determine the value of hypoosmotic swelling test in the prediction of sperm viability. For this purpose, hypoosmotic swelling and eosin Y were performed in parallel and in combination, on both fresh and freeze-thawed semen. Rates for eosin Y were significantly higher than for the hypoosmotic swelling test in fresh semen, with a weak, though significant correlation between the two tests (r = 0.47, p < 0.05). When both tests were performed in succession (hypoosmotic swelling test followed by eosin Y), 14.6% of swollen sperm incorporated the dye. Following exposure to hypoosmotic conditions, sperm viability decreased by 35%. When sperm were killed by freezing, hypoosmotic swelling test rates were higher than eosin Y. Results indicate that these two tests cannot be used interchangeably, since 15% of the swollen sperm apparently died, suggesting that plasma membrane integrity is lost before the capacity to maintain osmotic equilibrium.

PCR analysis of Y-chromosome sequences in a 45,X male patient and a review of the literature.

Abstract: The 45,X karyotype is usually associated with Turner syndrome, while male phenotype is exceptional. The authors report a 45,X male patient with normal external genitalia and sex behavior, but who was azoospermic. He had a normally developed musculature and pilose distribution, testicular volume of 15 mL and no gynecomastia but clinical stigmata of Turner syndrome (short stature, short neck and 4th metacarpal bones) and azoospermia. Hormonal plasma levels of testosterone, estradiol, prolactin, and gonadotrophins were within the normal range as was the response of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) (30 and 60 min) after 100 mug iv of LH-RH administration. Testicular biopsy could not be performed. Karyotype was 45,X without evidence of mosaicism. Polymerase chain reaction of genomic DNA studied with 12 different sequences of Y chromosome revealed only the presence of SRY gene (testis determining factor). It is possible that SRY/autosomal translocation had occurred in this patient...

HIGH-AFFINITY SITES FOR beta-D-GALACTOSIDASE ON MEMBRANE-BOUND VESICLES ISOLATED FROM RAT EPIDIDYMAL FLUID

Abstract: Glycosidases in rat epididymal fluid are secreted under androgen stimulation and possess receptors on the sperm surface. One of these enzymes, beta-D-galactosidase (gal), was found in the epididymal fluid as a soluble enzyme and also in a heterogeneous population of membrane bound vesicles (mbv). beta-D-Galactosidase was specifically localized to a subpopulation of larger, electron-dense mbv. The aim of this study was to analyze the high-affinity sites for gal on the membrane of mbv using two different methods: classical fluorometric assay (used in previous papers) and colloidal gold (20 nm) conjugated to gal as a marker in ultrastructural studies. beta-D-Galactosidase bound to mbv with high-affinity (Kd in a nanomolar range) are in a saturable form. Furthermore, 25 mM fructose-1,6-diphosphate (f-1,6-dip), a sugar that competes for the binding site, showed 50% inhibition of the binding. The gold conjugates were mostly observed on the surface of the large, electron-dense mbv but not on the small, electronl...