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Testicular aspiration of sperm for intracytoplasmic sperm injection: a novel treatment for ejaculatory failure on the day of oocyte retrieval.

OBJECTIVE: To present a successful outcome of an IVF cycle complicated by failure to produce a sperm sample on the morning of oocyte retrieval. DESIGN: Case report. SETTING: Hospital-based IVF clinic. PATIENT: A couple suffering secondary infertility undergoing their first IVF and intracytoplasmic sperm injection (ICSI) cycle. INTERVENTION: Fine-needle testicular biopsy for sperm retrieval. MAIN OUTCOME MEASURES: Sperm retrieval, fertilization, pregnancy. RESULTS: Sufficient spermatozoa were obtained for ICSI. Ten metaphase II oocytes were injected with two embryos being transferred fresh and four cryopreserved. A single fetal heart was confirmed at 6-week sonar. CONCLUSION: Testicular aspiration of sperm for ICSI is a simple, inexpensive, and effective means of rescuing an IVF cycle in which the male partner unexpectedly has been unable to produce a sperm sample on the morning of oocyte collection.

Adult↗

Epididymal micropuncture with in vitro fertilization and oocyte micromanipulation for the treatment of unreconstructable obstructive azoospermia.

OBJECTIVE: To provide fertility to couples in whom the man has surgically unreconstructable obstructive azoospermia. DESIGN: Prospective. SETTING: Hospital-based IVF unit, including associated division of urologic microsurgery. PATIENTS: Couples referred to our fertility unit for treatment of men with surgically unreconstructable reproductive tract obstruction, including congenital absence of the vas deferens. MAIN OUTCOME MEASURES: Fertilization, pregnancies, and live births. RESULTS: Of 51 cycles in which sperm and eggs were retrieved, 67% (34/51) resulted in fertilization and 27.5% (14/51) developed clinical pregnancy. Clinical pregnancy rate per couple was 33% (14/43). A total of 15 live births have been obtained in 11 couples with one ongoing pregnancy. Epididymal length was the best predictor of sperm quality and pregnancy results. For couples with at least the corpus epididymis present, 41% (9/22) of cycles resulted in clinical pregnancies. CONCLUSIONS: Pregnancy rates are optimized using sperm retrieved from the epididymis by micropuncture and when micromanipulation is available for use during IVF.

Cell Survival↗

Birth of a healthy male after frozen thawed blastocyst transfer following intracytoplasmic injection of frozen thawed testicular spermatozoa from a man with nonmosaic Klinefelter's syndrome.

We here report on a birth of a male, with normal karyotype, after frozen thawed blastocyst transfer following intracytoplasmic injection of frozen thawed testicular spermatozoa from an azoospermic 27-year-old man with nonmosaic Klinerfelter's syndrome. Testicular sperm were retrieved by percutaneous needle biopsy.

Adult↗

Microsurgical vasoepididymostomy in the treatment of occlusive azoospermia.

OBJECTIVE: To evaluate the practice and outcome of the end-to-end and end-to-side techniques of vasoepididymostomy for the treatment of occlusive azoospermia. PATIENTS AND METHODS: This retrospective study comprised 31 unselected patients (mean age 32 years, range 22-59) operated on with one of two techniques, more usually a modified end-to-side technique. Sperm quality and paternity were assessed in 24 patients during a 2-year follow-up. RESULTS: Of the 15 patients in whom sperm were found at anastomosis, sperm were retrieved from 11. Of 24 patients, 11 had patent anastomoses and paternity was achieved in three cases. CONCLUSION: Although performed by a proficient microsurgeon the operative success rate was low, partly because the operation was technically demanding but largely because of the difficulty in assessing intraoperatively the level of occlusion and the quality of the sperm. Better techniques for the evaluation of sperm during the operation should be developed.

Adult↗

Chromatin defects in normal and malformed human ejaculated and epididymal spermatozoa: a cytochemical ultrastructural study.

Cytochemical defects in chromatin were examined by transmission electron microscopy (TEM) after the staining by alcoholic phosphotungstic acid (PTA) of normal and malformed ejaculated spermatozoa from 35 male partners of infertile couples, and in six sperm samples retrieved from the caput epididymidis of men affected by obstructive azoospermia. PTA staining was also analysed in normal ejaculates of fertile men after incubation of the washed spermatozoa with dithiothreitol (DTT) to reduce disulfides to thiols, or with DTT followed by iodoacetamide, a blocking agent for thiol groups. PTA stained 63 (27-100)% of malformed heads and 25 (10-100)% of normal sperm heads (median (range) n = 35; P = 0.0001, Wilcoxon matched pairs test). The percentage of normal heads stained by PTA was negatively correlated with the percentage of heads of normal form, with condensed chromatin and a normal acrosome (Spearman r = 0.75; P = 0.0001), and positively correlated with the percentage of malformed heads after conventional TEM analysis (Spearman r 0.60; P = 0.0001). Staining with PTA in normal heads was not correlated with the presence of non-condensed chromatin in otherwise normal sperm heads evaluated by conventional TEM analysis. In spermatozoa recovered from the caput epididymidis, 15% of normal heads were stained with PTA, significantly fewer than in ejaculated sperm samples (P = 0.014). The reduction of disulfides to thiols was associated with PTA staining of all normal heads, and this was prevented by incubation with iodoacetamide. We conclude that PTA staining of the nuclei of human ejaculated spermatozoa may indicate a defect of chromatin condensation, owing to an excess of free thiol groups. The lower percentage of normal epididymal sperm heads that stained with PTA in cases of obstructive azoospermia compared with ejaculated sperm may be related to an overoxidation of thils owing to the ageing of spermatozoa.

Chromatin↗

[MESA (microsurgical epididymal sperm aspiration) and IVF (in vitro fertilization). A therapy concept in treatment of male infertility].

Microsurgical epididymal sperm aspiration (MESA), or sperm microaspiration retrieval technique (SMART), in conjunction with in vitro fertilization is a successful therapy in male infertility. From November 1991 to March 1994 a total of 29 attempts at MESA with subsequent IVF were made. Of 48 aspirations, 37 were successful and 13 attempts at IVF were possible, 6 of which were successful with 10 subsequent embryo transfers. In all, 3 pregnancies were achieved and 1 boy was born. In conclusion, microsurgical spermaspiration in conjunction with in vitro fertilization is a way of treating male infertility with a chance of achieving paternity with the partner's own sperm; the chances are probably better with intracytoplasmatic sperm injection.

Embryo Transfer↗

Male infertility after surgery for imperforate anus.

Congenital abnormalities of the genitourinary tract often coexist, and cryptorchidism is common in patients who have had imperforate anus. Twenty men who had pull-through procedures for imperforate anus in infancy have been evaluated for infertility. Seven had coexisting renal abnormalities, 4 had had recurrent epididymitis, 3 had had bilateral orchidopexies (at age 7 to 12), 2 had spina bifida, and 1 had a pituitary adenoma. Seven had no ejaculate (aspermia), 11 were azoospermic, 1 was severely oligozoospermic, and 1 had a normal sperm concentration in a small volume of ejaculate. Both vasa were blocked in 5 men, and this appeared to be a result of the original operative procedure. One vas was blocked in another 7 patients who had abnormalities on the contralateral side; three had epididymal blocks after epididymitis, and four had congenital malformations associated with an absent or ectopic kidney. After reconstruction (4), insertion of sperm reservoirs (4), microscopic epididymal sperm aspiration (2), or artificial insemination (1), sperm were retrieved from 9 men (ejaculated by 4) 2 pregnancies occurred. Male infertility after treatment of imperforate anus in infancy can be related to a wide variety of cause, some of which are amenable to treatment.

Abnormalities, Multiple↗

Postchemotherapy ejaculatory azoospermia: fatherhood with sperm from testis tissue with intracytoplasmic sperm injection.

PURPOSE: To define the success of testis sperm extraction (TESE) and intracytoplasmic sperm injection (ICSI) in azoospermic men with a history of chemotherapy. PATIENTS AND METHODS: In a retrospective study, 23 men with ejaculatory azoospermia and a history of chemotherapy underwent TESE in a search for usable spermatozoa. In six patients cryopreserved tissue and in nine patients fresh tissue provided sperm for an ICSI cycle. Histologic analysis of the testis was performed in all patients. The presence or absence of sperm, fertilization rates with ICSI, and final outcomes of pregnancy were recorded. RESULTS: Spermatozoa were found on TESE in 15 (65.2%) of 23 men. On histopathology, the predominant pattern observed was Sertoli cell only (47.8%), followed by hypospermatogenesis (30.4%), mixed (17.4%), and late maturation arrest (4.3%). The fertilization rate was 65.2%, and ongoing/delivered pregnancies occurred in 30.8% of cycles. Six healthy boys and four healthy girls have been born to date. CONCLUSION: Men who are azoospermic and have had prior cytotoxic therapy make up a small subgroup of males with nonobstructive azoospermia. It is important to define and characterize this subgroup and better define their true fertility potential. Approximately two thirds of these men have retrievable testis sperm, which may be used with ICSI to have healthy offspring. This exciting avenue for paternity has heretofore not been available to such patients.

Adolescent↗

Preimplantation genetic diagnosis for beta-thalassaemia: the Sardinian experience.

OBJECTIVES: To report the experiences on preimplantation genetic diagnosis (PGD) in couples at risk for beta-thalassaemia in Sardinia. METHODS: 23 couples at risk for beta-thalassaemia were included in the PGD programme with a total of 42 cycles performed. Among these, 11 couples were fertile, while the remaining 12 had associated fertility problems. In vitro Fertilization (IVF), PGD and prenatal genetic molecular confirmation protocols and results are reported. RESULTS: All the patients followed the protocol of ovarian stimulation, oocyte retrieval, intracytoplasmic sperm injection (ICSI), embryo biopsy and genetic analysis. A total of 272 oocytes were fertilized in the regular way, and embryo biopsy was performed on 202 embryos. Out of these 202 embryos, 192 (95%) were successful. The genetic diagnosis was performed on 150 embryos (78.1%). Ninety-eight were identified as unaffected and 75 were transferred in 31 cycles. In the infertile patient group, two biochemical pregnancies (11.1% per transfer), in the fertile patient group, four clinical pregnancies, two twin and two singleton pregnancies (30.8% per transfer), were obtained. The genetic molecular results were confirmed in all pregnancies by first-trimester chorionic villus sampling (CVS). CONCLUSION: Our study shows that PGD for beta-thalassaemia is an available procedure for couples who wish to avoid termination of pregnancy, except in cases where the IVF cycle efficiency is very poor.

Biopsy↗

Influence of staining and sampling procedures on goat sperm morphometry using the Sperm Class Analyzer.

Computer-assisted sperm morphometry analysis (ASMA) has improved the assessment of sperm morphology, but the results depend on the use of adequate sampling and staining procedures of spermatozoa from individual species. In this study, the Sperm Class Analyzer ASMA system was used for the morphometric analysis of goat sperm heads. Semen samples, obtained from four bucks, were used to evaluate the influence of three staining procedures (Diff-Quik, Hemacolor and Harris' Haematoxylin) on the accuracy of image processing and sperm morphometry, the effect of the number of cells analysed and the repeatability of the method. These experiments were performed to obtain objective, accurate and reliable sperm morphometric measurements of goat spermatozoa. Diff-Quik and Harris' Haematoxylin were significantly (p<0.05) more accurate than Hemacolor. However, Diff-Quik obtained the highest proportion of correctly analysed sperm heads (86.06%) and the lowest coefficients of variation on the image processing and morphometric measurements. The staining methods affected significantly the sperm dimensions (p<0.001) with increased values from Diff-Quik than Hemacolor and Harris' Haematoxylin, respectively (Diff-Quik>Hemacolor>Harris' Haematoxylin). No differences in morphometric parameters were found when 100, 150, 175 or 200 spermatozoa were analysed. The repeatability of results obtained was very high since no differences were found when measuring the same sperm on multiple attempts. In conclusion, to obtain objective, accurate and repeatable sperm morphometric measurements by the Sperm Class Analyzer system in goats, the analysis of 100 spermatozoa from slides which have been previously stained with Diff-Quik is recommended.

Animals↗

Successful birth after intrafallopian transfer of microhatched embryos.

OBJECTIVE: To present a successful transfer of microhatched embryos to the fallopian tubes via microlaparoscopy. DESIGN: Case report. SETTING: Private practice affiliated with a medical university. PATIENT: A 40-year-old woman with primary infertility, mildly elevated baseline FSH levels, and a history of poor ovarian response to ovulation induction. Her husband had severe oligoospermia after vasectomy reversal. INTERVENTION(S): Late luteal leuprolide acetate to pituitary down-regulation followed by pure FSH, 300 IU, and hMG, 300 IU, daily for ovulation induction. Transvaginal oocyte retrieval, intracytoplasmic sperm injection, assisted embryo hatching, microlaparoscopic intrafallopian ET. MAIN OUTCOME MEASURE(S): Amniocentesis at the 14th week of gestation revealed a normal karyotype (46,XX), birth of a normal female infant (3700 g). RESULT(S): Establishment of a single, viable intrauterine gestation followed by a vaginal delivery at term. CONCLUSION(S): This case shows the possibility of using assisted-hatched embryos for laparoscopic intrafallopian tube transfer.

Adult↗

The development of an oocyte-containing follicle during gonadotrophin-releasing hormone agonist administration.

Administration of gonadotrophin-releasing hormone (GnRH) agonist in a 29 year old woman with infertility due to ovulatory dysfunction resulted in the development of several ovarian cysts. After human chorionic gonadotrophin (HCG) was injected, the cysts were aspirated and one mature oocyte was retrieved. Intracytoplasmic sperm injection (ICSI) was performed and the resulting embryo was transferred. A singleton pregnancy was obtained and a healthy baby was born at 36 weeks of gestation. Because GnRH agonist-derived cysts may contain oocytes, we suggest that when the growth of cysts is accompanied by high concentrations of oestradiol, the administration of HCG may be useful to achieve oocyte maturation and advance IVF treatment.

Adult↗

First recorded pregnancy and normal birth after ICSI using electrophoretically isolated spermatozoa.

BACKGROUND: DNA damage in the male germ line is associated with poor fertilization and cleavage rates, impaired embryo quality and early pregnancy loss. Given these associations, embryologists are keen to develop techniques that will allow the selection of viable spermatozoa exhibiting low levels of DNA damage for assisted conception purposes. METHODS: In this article, we describe a novel electrophoretic approach for the rapid isolation of cells possessing little DNA damage. The limits of the method were examined using cryostored and snap-frozen semen samples as well as testicular biopsy material. In addition, clinical utility was demonstrated in a case study involving treatment of a patient exhibiting persistently high levels of DNA damage in his spermatozoa. RESULTS: From a range of difficult starting materials (biopsies, cryostored semen and snap-frozen sperm suspensions), the electrophoretic system rapidly isolated populations of motile, viable, morphologically normal spermatozoa exhibiting high levels of DNA integrity. Clinical application in a couple suffering from long-term infertility associated with extensive DNA damage in the male germ line led to the first human pregnancy following such electrophoretic sperm isolation. CONCLUSIONS: The electrophoretic procedure holds promise as a convenient method for the rapid preparation of high-quality spermatozoa for assisted conception purposes.

Cell Separation↗

Can inhibin-B predict the outcome of microsurgical epididymal sperm aspiration in patients with suspected primary obstructive azoospermia.

AIM: To evaluate whether inhibin-B can predict the outcome of a microsurgical epidymal sperm aspiration (MESA) procedure in patients with suspected primary obstructive azoospermia (OA) and if inhibin-B can replace testicular biopsy in the diagnostic work-up of these patients. METHODS: Inhibin-B levels and testicular biopsy scores were related to the outcome of MESA in 43 patients with suspected primary OA. MESA was considered to be successful when epididymal sperm could be identified during the procedure. RESULTS: Spermatozoa were present in the epididymal aspirate in 28 out of the 43 patients (65%). Inhibin-B values were not significantly different in patients with successful or unsuccessful MESA. The modified Johnsen score, however, was significantly lower in patients with unsuccessful MESA (P = 0.003). A rete testis obstruction or epididymal malfunctioning was found in 15% of patients with suspected primary OA, reflected by unsuccessful MESA despite normal inhibin-B levels and normal testicular histology. CONCLUSION: Inhibin-B cannot replace testicular biopsy as a diagnostic tool in the work-up of patients with suspected primary OA. Testicular biopsy is useful in identifying patients with spermatogenic arrest, who might have normal inhibin-B values.

Azoospermia↗