Evaluation of spermatogenesis using flow cytometry and confocal microscopy.
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Age Factors↗
Biomedical subjects
Y Soffer
Publications and source records attributed to Y Soffer.
Factors influencing the outcome of ICSI in patients with obstructive and non-obstructive azoospermia: a comparative study.
BACKGROUND: Factors influencing success of sperm retrieval in azoospermic patients and outcome of ICSI were evaluated. METHODS AND RESULTS: Uni- and multifactorial analysis were performed using logistic and stepwise analysis, following surgical sperm retrieval by percutaneous epididymal sperm aspiration (55 cycles) or testicular sperm extraction (142 cycles) in 52 and 123 patients with obstructive azoospermia (OA) and non-obstructive azoospermia (NOA) respectively. ICSI cycles using fresh or cryopreserved-thawed sperm were included. Sperm were retrieved to allow ICSI in 100 and 41% of OA and NOA patients, with no significant correlation with patients' age or FSH level. Occurrence of pregnancy was significantly correlated with female age (90th quantile: 38 years), number of oocytes retrieved (10th quantile: five oocytes) and number of oocytes injected (10th quantile: four oocytes). Sperm origin (epididymal versus testicular), status (fresh or thawed), male partner's age, and serum FSH had no significant effect upon implantation rate, pregnancy rate per embryo transfer or spontaneous miscarriage rate. CONCLUSIONS: In OA patients ICSI should be planned in conjunction with surgical sperm retrieval. In contrast, the lack of efficient non-invasive parameters to predict sperm retrieval in NOA suggests that elective surgical sperm retrieval may be offered to these patients prior to ovarian stimulation of their partners, especially when donor back-up is not an alternative. Female factors such as age and ovarian reserve have significant impact upon clinical success rates.
Adult↗
Influence of a short or long abstinence period on semen parameters in the ejaculate of patients with nonobstructive azoospermia.
OBJECTIVE: To compare the effect of a short (4 days) or a long (14 days) abstinence period on sperm retrieval by extended sperm preparation in patients with nonobstructive azoospermia scheduled for testicular biopsy and intracytoplasmic sperm injection (ICSI). DESIGN: A prospective case control study. SETTING: Male infertility clinic in a university hospital. PATIENT(S): Fifty male patients with nonobstructive azoospermia, scheduled for testicular biopsy for ICSI. INTERVENTION(S): Diagnosis of nonobstructive azoospermia and a thorough microscopic search for sperm cells (extended sperm preparation). MAIN OUTCOME MEASURE(S): The number of sperm cells collected, sperm motility, and total motile sperm count after short and long abstinence periods. RESULT(S): There was a significant difference between long and short abstinence with an increase in sperm count (log-to-log transformed analysis of variance P<.025) and total motile sperm (P<.025 analysis of variance, P<.02 paired Student's t-test) in the former group, but no significant change in sperm motility (Wilcoxon and paired Student's t-test). In 18 patients, sperm concentration and sperm motility were similar in a second collection, done after the same abstinence period, compared with the same parameters in the first sample. When at least 10 motile sperm were defined as the cutoff number, allowing ICSI without testicular biopsy, no significant differences were found between the two abstinence periods. No clinical or laboratory male characteristic could predict the detection of 10 motile sperm by extended sperm preparation either after a short or a long abstinence period. CONCLUSION(S): Sperm count and total motile sperm were increased after a long abstinence period, with no change in sperm motility. No additional advantages were conferred by long abstinence as opposed to short abstinence when 10 motile sperm were defined as the cutoff number for ICSI. The recommended period of abstinence for extended sperm preparation and ICSI, whether short or long, should be individualized for each patient.
Adult↗
In vitro susceptibility of recent clinical isolates of Chlamydia trachomatis to macrolides and tetracyclines.
We tested the in vitro activity of clarithromycin, azithromycin, roxithromycin, erythromycin, doxycycline, and tetracycline against 50 clinical isolates of Chlamydia trachomatis. The minimal inhibitory concentrations (MICs) and the minimal bactericidal concentrations (MBCs) were determined in a tissue culture system using cycloheximide treated McCoy cells. MIC values for all the isolates were < or =0.015 microg/ml for clarithromycin, < or =0.125 microg/ml for roxithromycin and azithromycin, and < or =0.25 microg/ml for erythromycin and doxycycline. Almost half of the isolates (44%) were inhibited only by a concentration of 0.5 microg/ml of tetracycline. MBC as high as 4 microg/ml was displayed by doxycycline and tetracycline against 8% and 4% of the isolates respectively of the agents recommended by the Center for Disease Control as drugs of choice for the treatment of chlamydial infections, azithromycin exhibited a markedly better in-vitro activity than did erythromycin and doxycycline.
Anti-Bacterial Agents↗
Outcome of surgical treatment for superficial dyspareunia from vulvar vestibulitis.
OBJECTIVE: To evaluate the results of surgical treatment for superficial dyspareunia, as manifested by patient satisfaction, as well as epidemiologic characteristics of women with this medical problem. STUDY DESIGN: A questionnaire was sent to 69 women six months after the operation. It included questions about treatment before surgery and the impact of pain on the sexual relationship before and after the operation. Demographic, social and general health data were recorded before the operation. All patients returning the questionnaire were examined. RESULTS: Fifty-four (78%) patients replied. Half of those abstained from sexual relations before surgical treatment. Sixty-seven percent of patients required more than six visits to various physicians, before vestibulitis was diagnosed. Prior to surgery, 80% of patients received conservative treatment, whereas after surgery only 34% required it. A moderate to excellent improvement was reported after surgery by 45 (83%) patients. Repeat surgery (n = 7) resulted in further improvement in four patients. There were no major operative complications. Forty-five patients (83%) were satisfied with the results and would recommend the surgery to other women with this clinical problem. CONCLUSION: Surgical treatment for superficial dyspareunia from vestibulitis is quite safe and results in a high rate of patient satisfaction.
Adult↗
Sperm ultramorphology as a pathophysiological indicator of spermatogenesis in males suffering from varicocele.
Varicocele of spermatic veins is considered to be one of the major causes of male infertility associated with reduction of sperm quality. The pathophysiology of this condition is not yet completely understood. The aim of this study was to shed light on the pathophysiology of varicocele by identifying semen parameters, especially sperm ultramorphology, which improve following high ligation of the spermatic vein. Seventy-five males with diagnosed varicocele were included in this study. Semen parameters were assessed prospectively using light microscopy, semen biochemistry and sperm quantitative ultramorphological analysis, before high ligation and 3-9 months after high ligation. The control group consisted of twenty-five untreated varicocele patients who underwent two semen examinations within 3-9 months. No statistical difference in any of the examined variables was found between the two examinations in the control group. The treated patients exhibited a significant improvement in sperm density, progressive motility, percentage of normally formed spermatozoa, agenesis of sperm acrosome, chromatin condensation and incidence of amorphous heads compared with the pretreatment condition (P < or = 0.01). In contradiction, no significant improvement was observed following treatment in any of the sperm tail subcellular organelles. It is concluded that varicocele may cause deleterious alterations in early spermatid head differentiation during spermiogenesis and that varicocele patients with a high incidence of sperm acrosome and nucleus malformations are appropriate candidates for varicocele correction.
Adult↗
ART success and in vivo sperm cell selection depend on the ultramorphological status of spermatozoa.
Management of male infertility has recently shifted from treatment of the subfertile man towards techniques of assisted reproduction (ART). This study aimed to evaluate the possible role of the ultramorphological status of the spermatozoon with respect to sperm selection in vivo and prediction of ART success. Ultramorphological sperm parameters were assessed retrospectively for 92 males with sufficient sperm density (10(7) spermatozoa ejaculate-1) whose wives conceived following a stepwise discarding of the female genital tract barriers, using intra-uterine insemination (IUI) (n = 26), in vitro fertilization (IVF) (n = 45) or intracytoplasmic sperm injection (ICSI) (n = 21). In parallel, sperm samples of 71 fertile males were examined. Normal ultramorphology of all head and tail subcellular organelles was found to be essential for the ability of spermatozoa to pass the lower female genital tract. The ultramorphological migration threshold for this barrier is apparently higher than that essential for oocyte fertilization. No specific indication associated with passage through the upper genital tract was found. A high prevalence of axonema defects was found to impair the ability of sperm cells to penetrate the oocyte investment. The natural fertility index, based on routine sperm parameters and the ultrastructural status of the spermatozoon's subcellular organelles was confirmed to be beneficial for directing patients to ART. A discriminative score based on axonema integrity was found to contribute additional information for the first choice decision between conventional ART and ICSI (75% prediction ability). Thus it may be helpful in finding the simplest and least expensive procedure with the greatest long-term chance for pregnancy.
Female↗
Pregnancy and birth after transfer of embryos that developed from single-nucleated zygotes obtained by injection of round spermatids into oocytes.
OBJECTIVE: To use injection of spermatids into oocytes as a mode of infertility treatment in cases in which spermatozoa are not available. DESIGN: Prospective clinical evaluation and case report. SETTING: In Vitro Fertilization Unit, Herzliya Medical Centers, Herzliya-on-Sea, Israel. PATIENT(S): Thirteen couples with male factor infertility in which the male partner lacked spermatozoa in the ejaculate or testicular biopsy samples. INTERVENTION(S): Round spermatid injection and elongated spermatid injection into oocytes. MAIN OUTCOME MEASURE(S): Evaluation of the rate of two-pronucleated and single-nucleated zygote development. RESULT(S): The rate of two-pronucleated zygote development after round spermatid injection and elongated spermatid injection was relatively low (27% and 36%, respectively). Single-nucleated zygotes develop more frequently after round spermatid injection and elongated spermatid injection (35% and 17%, respectively) than after intracytoplasmic sperm injection with mature spermatozoa. A normal pregnancy and childbirth resulted from the transfer of 4 cleaving embryos, each of which developed from a single-nucleated zygote in a round spermatid injection treatment cycle with ejaculated spermatids. CONCLUSION(S): Embryos derived from single-nucleated zygotes after spermatid conception can be viable and give rise to an ongoing clinical pregnancy and childbirth.
Adult↗
The outcome of intracytoplasmic injection of fresh and cryopreserved epididymal spermatozoa from patients with obstructive azoospermia--a comparative study.
The aim of our study was to compare the outcome of intracytoplasmic sperm injection (ICSI) with fresh and frozen-thawed epididymal spermatozoa retrieved by percutaneous epididymal sperm aspiration (PESA) or microepididymal sperm aspiration (MESA) from patients with obstructive azoospermia. A retrospective analysis of consecutive ICSI cycles was performed, comparing the outcome in 24 patients with obstructive azoospermia undergoing surgical sperm aspiration by MESA (7 cycles) or PESA (17 cycles). In 23 of 24 patients, excess spermatozoa were cryopreserved. Following thawing, 21 ICSI cycles were performed (11 cycles after MESA, 10 after PESA). No statistically significant differences were noted in all parameters examined in ICSI cycles with fresh or cryopreserved spermatozoa from the same patients. Comparing all ICSI cycles with fresh and frozen-thawed epididymal spermatozoa, the rates of two-pronuclear fertilization (56% versus 53%), embryo cleavage (90% versus 86%), implantation (10% versus 14%), clinical pregnancy per embryo transfer (32% versus 37%) and delivery/ongoing pregnancy rate (27% versus 26%) were not statistically different. The cumulative ongoing pregnancy rate per sperm retrieval procedure was 46%, respectively. We conclude that the clinical outcome of ICSI with fresh and frozen-thawed spermatozoa after retrieval by PESA was similar to that by MESA. Epididymal sperm cryopreservation in patients with obstructive azoospermia is feasible and efficient using a simple freezing protocol and should be offered to optimize the yield of pregnancies achieved following such procedures.
Adult↗
In-vitro human spermatozoa nuclear decondensation assessed by flow cytometry.
The process of sperm chromatin decondensation occurs when a spermatozoon enters an ovum. Protamine disulphide bonds are reduced to SH and the polycationic protamines combine with the polyanionic egg protein, nucleoplasmin, thus being stripped from DNA which then combines with histones. Defective chromatin decondensation will thus prevent further development of the male pronucleus. In this study human sperm samples were incubated in vitro at 28 degrees C (using a medium in which the polyanion, heparin, substitutes for nucleoplasmin and beta-mercaptoethanol for egg glutathione) for 10, 20 and 30 min before stopping the reaction with formalin (to 3.6%). The DNA of the fixed cells was stained with Acridine Orange by a one-step method and subjected to flow cytometry and data analysis, in which a zone characteristic of condensed chromatin is outlined on red-green fluorescence contour plots. After 20 min of incubation 97% of the control spermatozoa that were in the mature window (WIN M) had decondensed and moved out of this region. Defects in sperm decondensation were seen in four semen samples of the 20 that were tested. In cases where spermatozoa fail to produce a fertilized egg the cause may lie with defective chromatin quality, including failure of the sperm chromatin to decondense. The method described here is a simple procedure for detecting sperm samples containing such defective cells.
Animals↗
Intracytoplasmic injection of fresh and cryopreserved testicular spermatozoa in patients with nonobstructive azoospermia--a comparative study.
OBJECTIVE: To compare the outcome of intracytoplasmic sperm injection (ICSI) with fresh and frozen-thawed testicular spermatozoa in patients with nonobstructive azoospermia. DESIGN: Retrospective analysis of consecutive ICSI cycles. SETTING: In Vitro Fertilization Unit, Assaf Harofeh Medical Center. PATIENT(S): Eighteen with nonobstructive azoospermia in whom testicular sperm was found after testicular sperm extraction. INTERVENTION(S): Testicular sperm retrieval, cryopreservation, and ICSI with fresh or frozen-thawed testicular spermatozoa. MAIN OUTCOME MEASURE(S): Two-pronuclear fertilization; embryo cleavage rates, mean number of embryos transferred per cycle, and their relative quality, embryo implantation, clinical pregnancy, and ongoing pregnancy rates (PRs) per ET. RESULT(S): No statistically significant differences were noted in all parameters examined between ICSI cycles with fresh or cryopreserved testicular spermatozoa from the same nine patients and comparing all ICSI cycles performed; with fresh (25 cycles) and thawed (14 cycles) testicular spermatozoa, respectively: two-pronuclear fertilization, 47% versus 44%; embryo cleavage rates, 94% versus 89%; implantation rates, 9% versus 11%; and clinical PR, 26% versus 27%. The delivery or ongoing PR using fresh sperm was better (21% versus 9%), but the difference did not reach statistical significance. The cumulative clinical PRs and ongoing PRs per testicular sperm extraction procedure were 36% and 24%, respectively. CONCLUSION(S): Testicular sperm cryopreservation using a simple freezing protocol is promising in patients with nonobstructive azoospermia augmenting the overall success achieved after surgical sperm retrieval.
Adult↗
The outcome of in vitro fertilization in unexplained habitual aborters concurrent with secondary infertility.
OBJECTIVE: To compare the IVF-ET reproductive performance of unexplained habitual aborters with nonaborting women with similar characteristics also treated with IVF-ET. DESIGN: Controlled clinical study. SETTING: In Vitro Fertilization Unit, Assaf Harofeh Medical Center, Zerifin, Israel. PATIENT(S): Twenty patients with habitual abortions and secondary infertility were subjected to 42 IVF-ET cycles. Their reproductive performance in IVF-ET was compared with the performance of the next followed nonaborting patients, pulled from a database, based on similar age and IVF indications. INTERVENTION(S): Ovulation induction with IM Depo-triptorelin 3.75 mg and menotropins. Oocyte retrieval, embryo transfer, and luteal supplementation with hCG. MAIN OUTCOME MEASURE(S): Mean number of oocytes and fertilizations, mean number of embryos and their morphology. Pregnancy rates (PRs) and pregnancy outcome. RESULT(S): The mean number of oocytes, fertilizations, and embryos was higher in the aborters compared with the controls. Morphology of the transferred embryos was similar in both groups though in general better quality embryos developed in the aborters group. Pregnancy rates were similar in both groups (32% and 29% per cycle). Pregnancy outcome of the habitual aborting patients in IVF-ET remained unsuccessful, as before treatment: 50% abortions (7/14) compared with only an 8% abortion rate (1/12) in the controls. CONCLUSION(S): Habitual aborters' performance at early stages of IVF-ET is better compared with their controls. Despite similar PRs in IVF-ET cycles, their pregnancy outcome after IVF-ET is unsuccessful, as before IVF-ET.
Abortion, Habitual↗
Extended sperm preparation: an alternative to testicular sperm extraction in non-obstructive azoospermia.
Testicular sperm retrieval for the treatment of non-obstructive azoospermia requires the execution of an invasive procedure, with all its possible attending complications and subsequent long-term effects. This study suggests a new non-invasive approach for collection of spermatozoa in these patients: the extended sperm preparation (ESP). ESP consists of conducting a thorough microscopic search through many droplets of ejaculate sediment. ESP was performed for 49 patients; in 17 patients (35%), spermatozoa were found and subsequently used in intracytoplasmic sperm injection (ICSI). Of these preparations, five yielded fewer motile spermatozoa than the number of corresponding oocytes available, and in one patient only non-motile spermatozoa were recovered. The remaining 32 ESP-negative patients underwent testicular sperm extraction (TESE) from testicular biopsy. Spermatozoa were found in 16 of 32 biopsies (50%) and subsequently used in ICSI. Fertilization and cleavage rates were comparable in both ESP and TESE groups, yielding four clinical pregnancies in each group (27 and 29% respectively). Embryo morphology was defined as excellent in significantly more cases in the ESP group than the TESE group, and implantation rate appeared somewhat higher in the ESP group (16%) than the TESE group (13%). The ESP technique yields results similar to TESE, and can be applied in cases of non-obstructive azoospermia as a prerequisite modality enabling us to avoid testicular biopsy in 35% of cases.
Adult↗
Testicular sperm retrieval by percutaneous fine needle sperm aspiration compared with testicular sperm extraction by open biopsy in men with non-obstructive azoospermia.
The efficiency of testicular sperm retrieval by testicular fine needle aspiration (TEFNA) was compared with open biopsy and testicular sperm extraction (TESE), in 37 rigorously selected patients with non-obstructive azoospermia. All patients underwent TEFNA and TESE consecutively. Thus, each patient served as his own control. The case was regarded as successful if at least one testicular spermatozoon was found allowing intracytoplasmic sperm injection (ICSI) of at least one oocyte. The mean age of the male patients was 32.7 years (range 24-47). Whereas by TEFNA spermatozoa enabling performance of ICSI were found in only four patients out of 37 (11%), open biopsy and TESE yielded spermatozoa in 16 cases (43%). The negative predictive value of high serum follicle stimulating hormone (FSH) concentrations (> or =10 IU/l) (predicting failure to find spermatozoa for ICSI) was low (38.4%). The positive predictive value (predicting the chance to find spermatozoa for ICSI) of normal-sized testicle was not different from that of small-sized (<15 ml) testicle (50%). Complications included one case of testicular bleeding following fine needle aspiration, treated locally, and two cases of extratunical haematomata following TESE requiring no intervention. In patients with non-obstructive azoospermia, TEFNA has a significantly lower yield compared to TESE. Performance of ICSI with testicular sperm in these cases resulted in satisfactory fertilization and high embryo transfer rates. The implantation and pregnancy rates per embryo transfer were 13 and 29% respectively. Neither serum FSH values nor testicular size were predictive of the chances to find spermatozoa for ICSI. Some complications may occur even following TEFNA.
Adult↗
Evaluation of chromatin condensation in human spermatozoa: a flow cytometric assay using acridine orange staining.
The quality of sperm chromatin is an important factor in fertilization and is especially critical where one spermatozoon is artificially selected for fertilizing an egg (as in intracytoplasmic sperm injection). In this study, flow cytometry after staining of human spermatozoa with Acridine Orange was used to study chromatin structure. A method is described for estimating the percentage of cells in a human sperm sample that have completed epididymal maturation in regard to chromatin condensation. Of the 121 samples of the semen that were examined, nine contained a higher percentage of hypocondensed spermatozoa and six samples contained elevated amounts of hypercondensed spermatozoa. In addition to aberrancies in chromatin condensation other defects showed up as satellite populations of spermatozoa with higher than normal ratios of red/green fluorescence after Acridine Orange staining. Such defects were found in 15 semen samples. The use of swim-up and Percoll gradient centrifugation methods was shown to improve the percentage of spermatozoa with normal chromatin structure in some samples with poor initial quality.
Acridine Orange↗
Postejaculation serum prostate-specific antigen level.
OBJECTIVE: This study was carried out to determine whether ejaculation may modify the serum prostate-specific antigen (PSA) level and to investigate whether postejaculation serum PSA may play a role in male hypofertility work-up. METHODS: Serum PSA concentration was determined before and 1 h after ejaculation in 18 healthy men (group A) and in 16 men with male-factor infertility (group B). PSA change (delta) was recorded and analyzed. RESULTS: Postejaculation serum PSA differed significantly from basal levels (p = 0.0037 by the Wilcoxon signed rank test). Following ejaculation, a rise was noticed in 74% of subjects. The median relative change in PSA concentration was 54%. No difference in respect to mean, median and relative change was observed between study groups. In two cases (6%), PSA rose from normal level to more than 4 ng/ml. CONCLUSION: A significant postejaculation serum PSA elevation does occur, it is thus recommended that men abstain from ejaculation for 24 h prior to PSA sampling. Postejaculation PSA was not found to significantly correlate with hypofertility.
Adult↗
The benefits of mid-luteal addition of human chorionic gonadotrophin in in-vitro fertilization using a down-regulation protocol and luteal support with progesterone.
Luteal support is essential in in-vitro fertilization (IVF) when long-acting gonadotrophin-releasing hormone agonist (GnRHa) is used. Because progesterone lacks luteotrophic stimulation, it seems to be the drug of choice in cases with an increased risk of ovarian hyperstimulation syndrome (OHSS). The aim of this study was to assess the beneficial effect of the mid-luteal addition of human chorionic gonadotrophin (HCG) in IVF, using a down-regulation protocol and luteal support with progesterone, in a prospective randomized study. The study included 170 IVF cycles down-regulated with long-acting GnRHa which were supported with 50 mg/day progesterone i.m. during the luteal phase. Patients were evaluated in the mid-luteal period. Those without clinical signs of OHSS, oestradiol concentrations <1000 pg.ml and progesterone concentrations <50 ng/ml were randomly allocated to either the addition of 2500 IU HCG (HCG+ group) or no HCG (HCG- group). End luteal phase progesterone concentrations among non-pregnant patients were used to assess the contribution of exogenous progesterone and to categorize pregnancies according to their corpus luteum function. Similar low OHSS (2.7 and 1.8%) and pregnancy (30 and 29%) rates were observed in the HCG+ and HCG-groups respectively. Of the 26 pregnancies in the HCG+ cases, there was only one case with reduced corpus luteum function, compared with 12 or the 25 pregnancies among HCG-patients. Cases with reduced corpus luteum function required continuous progesterone support and presented lower betaHCG concentrations and a higher rate of adverse pregnancy outcome. We conclude that mid-luteal HCG addition does not affect pregnancy rate, but in fact helps to preserve corpus luteum function and avoids the need for further supplementation during early pregnancy.
Adult↗
Immunosuppressive effect of uterine fluid on spermatozoa-stimulated mixed lymphocyte cultures.
Spermatozoa, obtained from 20 healthy individuals, were incubated with crude uterine fluid or with a purified IgG fraction obtained from uterine fluid before adding them to cultures with allogeneic lymphocytes. Untreated spermatozoa induced a proliferative 1.5- to 6-fold increase, while the spermatozoa treated with crude uterine fluid or with purified IgG fraction caused 84-100% suppression of lymphocyte blastogenesis. These results suggest the existence of an endogenous uterine fluid factor, apparently IgG, which causes inhibition of lymphocyte proliferation induced by spermatozoa in the uterine lumen.
Adult↗