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Giant hydronephrosis of a duplex system associated with ureteral ectopia: a cause of retrograde ejaculation.

Retrograde ejaculation is an extremely rare complication of the ectopic ureter or ectopic ureterocele. In the literature, only two cases have been reported worldwide. In this article the authors describe a patient who has a completely duplicated collecting system with massive dilatation of the upper pole system, which has manifested as a huge abdominal and retroperitoneal mass, complicated by retrograde ejaculation.

Adult↗

Comparison between chromatin condensation and morphology from testis biopsy extracted and ejaculated spermatozoa and their relationship to ICSI outcome.

A significant association between male subfertility, imperfect spermiation and abnormal nuclear condensation has been suggested. The DNA content of spermatozoa might be responsible for inducing alterations in sperm morphology. The final nuclear shape, which is species-specific, depends on chromatin condensation during spermatogenesis as well as a precise organization of DNA within the nucleus. Many reports have described the association between disturbances in sperm chromatin condensation, morphology and male infertility. Chromatin condensation is achieved by gradual substitution of lysinerich somatic histones by testis-specific histone and finally by protamine. In this study two groups of patients were compared: the first consisted of 63 patients who had undergone intracytoplasmic sperm injection (ICSI) with freshly ejaculated spermatozoa whereas the second included 47 patients assigned to ICSI with testes biopsy-extracted spermatozoa. In both groups chromatin condensation was assessed by aniline blue staining and morphology evaluated according to strict criteria. The condensed chromatin and morphology of spermatozoa were significantly (P < 0.0001) less in the second group compared to the first. However the fertilization, cleavage, implantation and pregnancy rates were almost the same in both investigated groups. There was no significant difference between the two groups with respect to ICSI outcome. The percentage of chromatin condensation (nuclear maturity) and morphologically-normal spermatozoa were significantly higher (P < 0.0001) in the ejaculated spermatozoa than in those from testis biopsy but the ICSI outcome (fertilization, cleavage, implantation and pregnancy rates) was the same. In view of these results the fertilization capability and the embryo quality obtained using testis biopsy extracted spermatozoa is not influenced by chromatin condensation and sperm morphology in testicular sperm extraction (TESE)-ICSI programmes. Therefore, it could be said that neither chromatin condensation nor morphology of testis extracted sperm could predict the fertilization, implantation and pregnancy rate in TESE-ICSI programmes.

Adult↗

Frequency of hyper-, hypohaploidy and diploidy in ejaculate, epididymal and testicular germ cells of infertile patients.

The hypothesis that sperm aneuploidy and diploidy increase as a function of spermatogenesis impairment was addressed. Ejaculated semen samples from a series of men (n = 22) with very low total normal motile count (1 x 10(6)) was analysed in terms of sperm aneuploidy and diploidy by in-situ hybridization and compared with controls (n = 10). Germ cell aneuploidy was also analysed in an additional series of infertile patients presenting unexplained infertility (n = 3), congenital absence of the vas deferens (CAVD) (n = 6) and non-obstructive azoospermia (n = 3) undergoing IVF, microsurgical epididymal sperm aspiration (MESA)/ICSI and testicular sperm extraction (TESE)/ICSI cycles respectively. In-situ hybridization for chromosomes 1, 17, X and Y was performed on ejaculate, epididymal and testicular spermatozoa. Significantly higher sperm aneuploidy and diploidy rates where found (for the four chromosomes analysed) in spermatozoa from oligoasthenoteratozoospermia (OAT) over controls (18 versus 2.28% and 2.8 versus 0.13% respectively; P < 0.001). Testicular germ cells had even higher rates of sperm aneuploidy and diploidy. However, in this group it was difficult to determine whether the cells analysed were dysmorphic spermatozoa or spermatids. The data warrant further investigation on the cytogenetic abnormalities found in most germ cells identified in testicular tissue biopsies of azoospermic patients.

Adult↗

Birth of a healthy girl after ICSI with ejaculated spermatozoa from a man with non-mosaic Klinefelter's syndrome.

Klinefelter's syndrome is a major contributor to male infertility. Recent reports of births after ICSI with especially testicular spermatozoa from infertile men with this syndrome are promising. The birth of a healthy girl after ICSI treatment with ejaculated spermatozoa from a man with non-mosaic Klinefelter's syndrome is reported. The non-mosaic karyotype was confirmed by chromosome analysis of both peripheral blood leukocytes and fibroblasts from a skin biopsy. In conclusion, in a very few cases, men with apparently non-mosaic Klinefelter's syndrome have ejaculated spermatozoa that can result in a birth of a healthy child following ICSI.

Adult↗

Apoptosis and necrosis in human ejaculated spermatozoa.

BACKGROUND: Features of both apoptosis and necrosis have been reported in ejaculated human spermatozoa. This study examines the relative contribution of these two modes of cell death to the demise of these terminally differentiated cells. METHODS: Sperm fractions were prepared from aliquots of semen samples from young normozoospermic donors by simple washing from seminal plasma, by discontinuous density gradient centrifugation or by swim-up technique. They were subsequently incubated in vitro at 37 degrees C for 24 h. Sperm motility, viability, and the presence of two apoptotic markers, phosphatidylserine externalization (annexin-V binding) and DNA fragmentation (TUNEL), were examined before incubation and again after 4 and 24 h of incubation. RESULTS: The swim-up technique was the most efficient in terms of the recovery of viable, motile and non-apoptotic spermatozoa, followed by density gradient centrifugation and finally simple washing. No changes in the parameters tested were observed after 4 h of incubation, but a significant decrease in sperm motility and viability was detected after 24 h irrespective of the sperm preparation technique employed. However, these changes were not accompanied by any increase in the incidence of spermatozoa showing markers of an active apoptotic process. CONCLUSIONS: Healthy human ejaculated spermatozoa appear incapable of initiating apoptosis, at least under in vitro conditions.

Adult↗

Study of two markers of apoptosis and meiotic segregation in ejaculated sperm of chromosomal translocation carrier patients.

BACKGROUND: To try to explain the infertility of chromosomal translocation carrier patients, we compared the expression of two markers of apoptosis in the sperm of patients and of fertile donors, and we studied the meiotic segregation in the ejaculated sperm of these translocation carriers. METHODS: Twenty semen samples of translocation carriers, [reciprocal (n=14) and Robertsonian translocations (n=6)], were compared with the semen samples of donors (n=20). Different tests were applied: annexin V binding assay; terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labelling (TUNEL); and fluorescence in situ hybridization (FISH). RESULTS: The annexin V binding assay in sperm of patients with chromosomal translocation (n=17) showed a significantly increased proportion of sperm with externalized phosphatidylserine (PS) than in the control group (n=20, P<or=0.05). The rates of DNA fragmentation investigated by TUNEL reaction were higher in samples of translocation carriers (n=14) than in donors (n=20, P<0.0001). The measures by FISH technique showed that the proportions of balanced or normal gametes were predominant in the reciprocal translocation group (alternate: n=7; from 33.0 to 58.8%; adjacent I: n=7; from 4.6 to 43.8%) and in the Robertsonian translocation group (normal: n=5; from 76.0 to 88.5%). CONCLUSIONS: Our data show a predominant proportion of balanced gametes in sperm of chromosomal translocation carrier patients. Moreover, PS externalization and DNA fragmentation rates are significantly higher in ejaculated sperm of these patients than in donor sperm. These tests could be used to predict the outcome of ICSI for these patients.

Annexin A5↗

The expression of alpha v, alpha 5, beta 1, and beta 3 integrin chains on ejaculated human spermatozoa varies with their functional state.

Evidence has been presented suggesting the involvement of integrins and their ligands in mammalian fertilization. In this study we asked whether the alpha 5, alpha v, beta 1 and beta 3 integrin chains, which form receptors for fibronectin and vitronectin, are present on human spermatozoa. Fresh ejaculate spermatozoa and capacitated spermatozoa, before and after a calcium ionophore (A23187)-induced acrosome reaction, were either fixed and their reaction with anti-integrin monoclonal antibodies detected by immunoperoxidase staining or studied without fixation, using cytofluorimetric scanning. Expression of specific integrin chains varied with the functional state of spermatozoa. The alpha 5 chain was not detected on fresh living spermatozoa, but was present on capacitated spermatozoa, whether fixed or living. The pattern of beta 1 expression on living spermatozoa paralleled that of alpha 5. No further increase in the expression of either alpha 5 or beta 1 was observed following an ionophore-promoted acrosome reaction. In contrast, alpha v was detected on neither fresh, living ejaculate spermatozoa, nor following capacitation (< 10% reactive). The percentage of alpha v positive cells increased substantially following ionophore exposure. Expression of beta 3 was similar to alpha v, and the percentage of cells displaying beta 3 correlated with the proportion of spermatozoa that had undergone an acrosome reaction, following ionophore exposure. These results indicate that the expression of integrins on spermatozoa is dynamic, varying with their functional state and that integrin receptors for fibronectin (alpha 5 beta 1) become apparent on the spermatozoan surface during capacitation and vitronectin (alpha v beta 3) following the acrosome reaction.

Acetone↗

Detection of mRNA transcripts of beta 1 integrins in ejaculated human spermatozoa by nested reverse transcription-polymerase chain reaction.

Matrix proteins (e.g. fibronectin, laminin and vitronectin) and their receptors, especially beta 1 integrins, may play a role in sperm-oocyte interactions. These proteins are well described on spermatogenic cells but especially in ejaculated spermatozoa, the presence of mRNA as a necessity for their synthesis is not clear. Therefore the mRNA synthesis of the beta 1, alpha 3, alpha 4, alpha 5 and alpha 6 chains (fibronectin and laminin receptors) of the beta 1 integrins was investigated by nested reverse transcription-polymerase chain reaction (RT-PCR) in spermatozoa of human semen samples (n = 18) which showed normal classical semen parameters after swim-up preparation. After the second amplification, we obtained specific signals for mRNA transcripts of the beta 1, alpha 3, alpha 4 and alpha 6 chains in 17 of 18 semen samples investigated. All control PCRs were negative for somatic cell transcripts and contamination by somatic cells was excluded. These results demonstrate for the first time that human ejaculated spermatozoa contain the mRNA transcripts of beta 1 integrins and support the role of these proteins in adhesive mechanisms in the fertilization process.

Actins↗

Detection of mRNA transcripts of cyclic nucleotide phosphodiesterase subtypes in ejaculated human spermatozoa.

Multiple types and subtypes, including splice variants, of cyclic nucleotide phosphodiesterases (PDE) have been shown to be expressed in various tissues and organs. They control the intracellular level of cyclic nucleotides and are involved in hormonal signalling. In human spermatozoa, PDE play an important role in the regulation of motility, capacitation and acrosome reaction. The aim of this study was to investigate which transcripts of the different PDE types and subtypes could be found in human spermatozoa using reverse transcription-polymerase chain reaction (RT-PCR). Ejaculated spermatozoa from 10 single semen samples as well as another three semen sample pools were separated by swim-up and were investigated by RT-PCR. We obtained PCR products of the PDE types/subtypes 1A/B/C, 2, 3A/B, 4A/B/C, 5, and 8 with different intensities. Control PCR for leukocyte contamination were negative and contamination by other somatic cells was excluded by the spermatozoa preparation protocol, immunohistochemistry and visual examination. These results demonstrated for the first time that human ejaculated spermatozoa contain an extended pattern of PDE mRNA transcripts.

3',5'-Cyclic-AMP Phosphodiesterases↗

Caspase activity and apoptotic markers in ejaculated human sperm.

The objectives of this study were to determine if human ejaculated sperm exhibit active caspases and if caspase-dependent apoptosis markers are identifiable. Sperm from fertile donors and infertile patients were examined after gradient separation into leukocyte-free fractions of high and low motility. Sperm were evaluated for motion parameters, morphology, caspase activation, and apoptosis markers including phosphatidylserine (PS) translocation (annexin V binding) and DNA fragmentation (TUNEL). Active caspase-3 was detected by immunofluorescent microscopy in a small proportion of sperm in situ, in fractions of high and low motility sperm of patients and donors, but low motility fractions had significantly higher numbers of positive sperm. Immunoblot analysis detected inactive procaspase-3 (32 kDa) in all fractions of low sperm motility from patients and donors, while active caspase-3 (17 kDa) was only detected by immunoblotting in a limited number of low motility fractions from patients and in even fewer fractions from donors. Caspase enzymatic activity, as measured using the fluorogenic substrate DEVD-afc, was higher in patients than in donors in both low and high motility fractions. Annexin V staining and DNA fragmentation were detected in a proportion of sperm, with a higher frequency in the low motility fractions. A significant positive correlation between in-situ active caspase-3 in the sperm midpiece and DNA fragmentation was observed in the low motility fractions of patients, suggesting that caspase-dependent apoptotic mechanisms could originate in the cytoplasmic droplet or within mitochondria and function in the nucleus. These data suggest that in some ejaculated sperm populations, caspases are present and may function to increase PS translocation and DNA fragmentation.

Adult↗

Somatic cell apoptosis markers and pathways in human ejaculated sperm: potential utility as indicators of sperm quality.

In this study we extended earlier work to determine whether sperm respond to somatic cell apoptotic stimuli and whether apoptotic phenotypes are significant indicators of human sperm quality. We evaluated ejaculated sperm from fertile donors and subfertile patients following purification of fractions of high and low motility. In unstimulated conditions, caspase enzymatic activity was higher in motile fractions from subfertile patients than in donors, and was higher in low motility fractions from both groups. Staurosporine, but not a Fas ligand or H2O2, significantly increased caspase activity, but only in high motility fractions. Procaspase-3, -7 and -9 and low levels of active caspase-3, -7 and -9 were identified by immunoblot analysis. Apoptosis-inducing factor (AIF) was present in all samples but poly ADP-ribose polymerase-1 (PARP-1) was not detected. Phosphatidylserine translocation was significantly increased only with H2O2 treatment. In ejaculates of both subfertile and fertile men, we demonstrated the presence and activation of several proteins that are key constituents of apoptosis-related pathways in somatic cells, which may serve as markers for sperm quality.

Adult↗

Identification of transcripts by macroarrays, RT-PCR and in situ hybridization in human ejaculate spermatozoa.

Round spermatids contain high levels of extremely varied mRNAs that are synthesized either throughout early spermatogenesis or during spermiogenesis from the haploid genome. Concomitantly, with major changes in the chromatin organization, arrest of transcription occurs at midspermiogenesis. However, previous investigations using RT-PCR have revealed the persistence of numerous and different transcripts in ejaculated spermatozoa. In the present study, a step-by-step analysis by means of macroarray hybridization, RT-PCR and in situ hybridization was performed to identify more accurately the different mRNA species found in the human ejaculated spermatozoa. The data showed an extended pattern of various transcripts encoding a diverse range of proteins involved in signal transduction and cell proliferation. For the first time, they demonstrated that mRNAs coding for the transcription factors NFkappaB, HOX2A, ICSBP, protein kinase JNK2, growth factor HBEGF and receptors RXRbeta and ErbB3 accumulate within the sperm nucleus. The origin and fate of the sperm transcripts remain subject to discussion.

Ejaculation↗

Effects of multiple ejaculations after extended periods of sexual abstinence on total, motile and normal sperm numbers, as well as accessory gland secretions, from healthy normal and oligozoospermic men.

Four healthy donors provided three ejaculates on one day to deplete their sperm reserves and also after abstention periods of 1, 2, 4, 7, 10 and 14 days. The percentage of spermatozoa that were normally formed, progressively motile and which swelled in hypotonic medium remained unchanged throughout this period and the increase in total numbers of viable spermatozoa for 10 days prompted a comparison of seven donors and six oligozoospermic men. Multiple ejaculations after 10 days of abstinence for the patients with low normal, motile sperm numbers increased the total number, number of normal and number of motile spermatozoa by a mean of 230, 370 and 520%, respectively.

Adult↗

Pregnancy after intracytoplasmic sperm injection of metaphase II oocytes with spermatozoa from a man with complete retrograde ejaculation.

Retrograde ejaculation is an uncommon cause of infertility, which has been treated successfully with different kinds of artificial reproduction technique, e.g. cervical cap artificial insemination by husband, intra-uterine and intraperitoneal insemination, standard in-vitro fertilization, pronuclear stage transfer and gamete intra-Fallopian transfer. All these techniques require a minimal number and motility of spermatozoa obtained after post-masturbation voiding. In some cases, only very few spermatozoa with very poor or no motility are found in the urine voided immediately after masturbation. In such a case, where no more than 14 spermatozoa were recovered over a 3 h search, intracytoplasmic sperm injection of metaphase II oocytes led to the development and replacement of three fair embryos, resulting in an ongoing twin pregnancy. This technique opens up perspectives for the treatment of men with complete retrograde ejaculation and quasi-azoospermic post-voiding specimens.

Adult↗

Immunocytochemical localization of lipocalin-type prostaglandin D synthase in the bull testis and epididymis and on ejaculated sperm.

Previously, we identified a 26-kDa fertility-associated protein in bull seminal plasma as lipocalin-type prostaglandin D synthase. The objective of the present study was to immunohistochemically localize this enzyme to the various cell types within the bull testis and seven subsegments of the epididymis, and on ejaculated sperm in order to gain further insight into its potential function in male reproduction. In the testis, immunoperoxidase staining was localized within the elongating spermatids and Sertoli cells of the seminiferous tubules, varying with the stage of the spermatogenic cycle. The highest level of staining occurred during stages III-VII. The cuboidal epithelial cells of the rete testis and efferent ducts were also immunoreactive. Expression of lipocalin-type prostaglandin D synthase was not uniform in the seven epididymal subsegments, suggesting a possible role in sperm maturation. In all epididymal regions, expression was limited to the epithelial principal cells; no immunoreactivity was apparent in other cell types. Lipocalin-type prostaglandin D synthase was strikingly localized in the caput epididymidis, while moderate to weak staining was observed in the remainder of the epididymis. Droplets of reaction product observed within the lumen increased progressively from the caput to cauda. Using fluorescence microscopy, we also localized lipocalin-type prostaglandin D synthase to the apical ridge of the acrosome on ejaculated sperm.

Animals↗

Premature ejaculation: the current literature.

PURPOSE OF REVIEW: The recent increase in research with regard to premature ejaculation has led to a significant number of new papers looking at the diagnosis, definition, aetiology and management of this condition. RECENT FINDINGS: The intravaginal ejaculatory latency time remains the primary measure of ejaculatory time although increasing bother and distress require assessment and establishment of quantifiable measures. Biological and psychogenic causes contribute to a multifactorial model of premature ejaculation with some neurobiological vulnerability. The principal treatments are selective serotonin reuptake inhibitors and behavioural cognitive interventions. New treatment interventions are under investigation. SUMMARY: A number of guideline papers confirm that a primary sexual history and a multimodel treatment approach provide the best approach to patients with this common condition.

Adult↗

Comparison of dapoxetine versus paroxetine in patients with premature ejaculation: a double-blind, placebo-controlled, fixed-dose, randomized study.

PURPOSE: To compare the efficacy and safety of dapoxetine, paroxetine, and placebo for the oral pharmacotherapy of premature ejaculation. MATERIALS AND METHODS: Three hundred forty potent men with premature ejaculation were recruited to this study. Patients were randomly assigned to receive 60 mg dapoxetine (group 1, n = 115), or 20 mg paroxetine (group 2, n = 113) or placebo (group 3, n = 112) orally daily during a 12-week period for each agent. The efficacy of the 3 treatments was assessed every 2 weeks during treatment and at the end of study using responses to International Index of Erectile Function (IIEF), intravaginal ejaculatory latency time (IELT) evaluation, mean intercourse satisfaction domain, mean weekly coitus episodes and adverse drug effects. RESULTS: At the end of the 12-week treatment with dapoxetine, paroxetine, and placebo, the mean IELT was increased from 38, 31 and 34 seconds to 179, 370 and 55 seconds, respectively (P = 0.01 in group 1 and P = 0.001 in group 2). Baseline mean intercourse satisfaction domain values of International Index of Erectile Function of 10, 11, and 11 reached 14, 17 and 12 at the end of the 12-week treatment in groups 1, 2, and 3 respectively (P = 0.03 in groups 1, 2). The mean weekly intercourse episodes increased from pretreatment values of 1.4, 1.3, and 1.3 to 2.2, 2.5 and 1.4, for dapoxetine, paroxetine and placebo, respectively (P = 0.04 in groups 1, 2). The incidence of adverse effects with dapoxetine and paroxetine was significantly higher (P = 0.04 in groups 1, 2) compared to that of placebo. CONCLUSIONS: Paroxetine appears to provide significantly better results in terms of IELT and intercourse satisfaction versus dapoxetine. Each treatment was well tolerated.

Adult↗

A comparative study of ejaculate traits in three endangered ungulates with different levels of inbreeding: fluctuating asymmetry as an indicator of reproductive and genetic stress.

We studied three closely related species of endangered gazelles (Gazella dorcas, Gazella dama and Gazella cuvieri) with different levels of inbreeding in order to determine at which intensities inbreeding influences ejaculate traits. We also examined whether fluctuating asymmetry (FA) is a reliable indicator of genetic as well as reproductive stress. Our results show that, within each population, the individual coefficient of inbreeding is inversely related to ejaculate quality only in the species with the highest levels of inbreeding (G. cuvieri). In addition, FA is a reliable indicator of individual levels of inbreeding in both the species with the highest levels of inbreeding (G. cuvieri) and the species with intermediate levels of inbreeding (G. dama). Thus, FA appears in individuals whose levels of inbreeding are still not high enough to affect male reproductive potential and should therefore be considered a sensitive indicator of genetic stress. Finally, FA is also a reliable indicator of male reproductive stress since it is related to individual semen quality in all the species studied.

Animals↗