Yigal Soffer

Intracytoplasmic injection of fresh and cryopreserved testicular spermatozoa in patients with nonobstructive azoospermia—a comparative study

OBJECTIVEnTo compare the outcome of intracytoplasmic sperm injection (ICSI) with fresh and frozen-thawed testicular spermatozoa in patients with nonobstructive azoospermia.nnnDESIGNnRetrospective analysis of consecutive ICSI cycles.nnnSETTINGnIn Vitro Fertilization Unit, Assaf Harofeh Medical Center.nnnPATIENT(S)nEighteen with nonobstructive azoospermia in whom testicular sperm was found after testicular sperm extraction.nnnINTERVENTION(S)nTesticular sperm retrieval, cryopreservation, and ICSI with fresh or frozen-thawed testicular spermatozoa.nnnMAIN OUTCOME MEASURE(S)nTwo-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.nnnRESULT(S)nNo 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.nnnCONCLUSION(S)nTesticular sperm cryopreservation using a simple freezing protocol is promising in patients with nonobstructive azoospermia augmenting the overall success achieved after surgical sperm retrieval.

Pregnancy and birth after transfer of embryos that developed from single- nucleated zygotes obtained by injection of round spermatids into oocytes

OBJECTIVEnTo use injection of spermatids into oocytes as a mode of infertility treatment in cases in which spermatozoa are not available.nnnDESIGNnProspective clinical evaluation and case report.nnnSETTINGnIn Vitro Fertilization Unit, Herzliya Medical Centers, Herzliya-on-Sea, Israel.nnnPATIENT(S)nThirteen couples with male factor infertility in which the male partner lacked spermatozoa in the ejaculate or testicular biopsy samples.nnnINTERVENTION(S)nRound spermatid injection and elongated spermatid injection into oocytes.nnnMAIN OUTCOME MEASURE(S)nEvaluation of the rate of two-pronucleated and single-nucleated zygote development.nnnRESULT(S)nThe 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.nnnCONCLUSION(S)nEmbryos derived from single-nucleated zygotes after spermatid conception can be viable and give rise to an ongoing clinical pregnancy and childbirth.

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.

Prevalence of genital chlamydia and mycoplasma infection in couples attending a male infertility clinic

To investigate the prevalence of genital mycoplasma (Ureaplasma urealyticum andMycoplasma hominis) andChlamydia trachomatis infections in infertile and fertile couples, 135 infertile couples from a male infertility clinic and 88 fertile couples from a prenatal care clinic were randomly screened. Semen, urethral and cervical smears were cultured for genital mycoplasma and chlamydia. Antichlamydia IgA, IgG and IgM antibodies were tested in blood and semen. Although the overall prevalence ofUreaplasma urealyticum was higher than that ofMycoplasma hominis, the prevalence ofMycoplasma hominis was significantly higher in the infertile men and women. The prevalence ofChlamydia trachomatis, positive culture was higher in infertile couples, but this difference was not significant. The prevalence of serum-specific IgG was significantly higher in infertile than in fertile women (11.9 vs 3.4%,p<0.05). Specific semen IgA was significantly higher in infertile than in fertile men (8.9 vs 1.1%,p<0.015). A low rate of concordance between partners was found inMycoplasma andChlamydia culture, and in the same patient between seminal IgA and urethral culture. In conclusion it is necessary to check both culture and serology findings in both partners.

Increased fertilization and pregnancy rate in polypronuclear fertilization cycles in in vitro fertilization-embryo transfer

OBJECTIVEnTo compare fertilization and pregnancy rates between cycles with polypronuclear fertilizations and cycles with normal fertilizations.nnnDESIGNnIn vitro fertilization-embryo transfer (IVF-ET) cycles in which oocytes were retrieved were divided into two groups according to the nature of fertilization.nnnPATIENTSnAll patients were participants of our IVF-ET program.nnnRESULTSnA significantly higher fertilization rate was found in the polypronuclear fertilization cycles (61% versus 36.6%) and also an improved pregnancy rate (47.5% versus 19.6%) and per embryo transfers (53% versus 28.8%). The vast majority of polypronuclear fertilizations occurred in mature oocytes.nnnCONCLUSIONnWe believe that the increased receptability of the oocytes improves fertilization and conception rates. The polypronuclear fertilization is an extreme expression of such improved receptibility and should be considered as an encouraging sign for conception.

Difficult ICSI Cases: An Israeli Survey

This chapter is based on a survey of published material from our center as well as from 12 other Israeli in vitro fertilization (IVF) centers during 1997. This material deals mainly with nonobstructive azoospermia, and allowed us to answer the following questions in difficult intracytoplasmic sperm injection (ICSI) cases: Is it possible to do ICSI in every nonobstructive azoospermia? How to retrieve sperm in difficult cases? Is ICSI feasible with nonmotile spermatozoa or with spermatids only? Is sperm cryopreservation contributory? Is it possible to overcome unexpected egg factor in ICSI?

Fertilizing Capability of Frozen-Thawed Immotile Sperm

The introduction of intracytoplasmic sperm injection (ICSI) for severe male cases has made it possible to treat patients with azoospermia, in whom sperm is achieved from the epididymis and the testis (1–3). In some of the treated male patients, especially those with non-obstructive azoospermia, mainly immotile spermatozoa are available. Because it is assumed that the fertilizing potential of testicular sperm is lower (4) and motility is an important predictive factor for successful fertilization (5) it should be questioned whether ICSI perfomed with immotile sperm is justified.

Studies on sperm antigenicity. 2. in vivo and in vitro cellular reactivity in guinea pigs sensitized to homologous and heterologous spermatozoal autoantigens*†*Supported by Grant 8865 from the Israeli Chief Scientist-Ministry of Health and by the French-Israel Treaty on Public Health Research (Centre National de la Recherche Scientifique).†Presented in part at the Second International Meeting of Immunology, July 1974, Brighton, England.

In vitro delayed type hypersensitivity was demonstrated with peritoneal exudate cells from guinea pigs of the Rockefeller and Hartley strains, immunized with different preparations of the guinea pig male reproductive tract (RMT) emulsified in complete Freunds adjuvant-H37Ra. The RMT preparations were purified guinea pig spermatozoal autoantigens S, P, and T; whole guinea pig spermatozoa, and extract from epididymal tissue. The cellular sensitivity in vivo was demonstrated by injecting the proper antigen into the skin of the tested animals and in vitro by the macrophage inhibition technique. Peritoneal exudate cells from guinea pigs sensitized with whole guinea pig spermatozoa cells were inhibited in vitro by the specific antigen, epididymal extract, and autoantigen-T. Autoantigen-S was found to be a weak immunogen. However, the migration of peritoneal exudate cells from guinea pigs sensitized with large amounts of antigen-S was inhibited by whole spermatozoa in vitro. This cross-reactivity revealed the possibility that the immunogenicity of purified autoantigen-S might be connected to its molecular size. According to the immunizing dose of the antigens, testicular lesions of either the aspermatogenic or orchitis type were found in the testes of sensitized guinea pigs. Lesions in the testes of the guinea pigs were not detectable by cross-immunization with heterologous human or rat spermatozoa, although some degree of in vitro cross-reactivity was detected by skin test studies.