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Biomedical subjects

M L Windt

Publications and source records attributed to M L Windt.

At least 19 recordsLinked to original sources
PubMed2002

Intracytoplasmic sperm injection with testicular spermatozoa in men with azoospermia.

PURPOSE: The aim of the study was to gain an insight into the optimal management of the infertile couple with the husband suffering from azoospermia. METHODS: One hundred and forty-two intracytoplasmic sperm injection (ICSI) cycles performed with testicular extracted spermatozoa were retrospectively analysed. The following factors were investigated for their possible influence on fertilization, cleavage, damage, pregnancy, and ongoing pregnancy rates: the use of fresh, cryopreserved, and preincubated (24 h) spermatozoa and the etiology of the husbands' azoospermia (obstructive and nonobstructive). All microinjections were performed with apparently normal spermatozoa--a head with a tail of normal length. In 116 cycles at least two embryos were available for transfer. RESULTS: The overall fertilization, clinical pregnancy, and ongoing pregnancy rates obtained for the 116 cycles were 65.0, 30.2, and 22.4% respectively. Similar outcomes were obtained for cycles using fresh testicular and cryopreserved testicular spermatozoa. Similarly, no significant differences were obtained between the cycles using spermatozoa from obstructive or nonobstructive azoospermic patients. An increase in motility after a 24-h preincubation was observed, and although this group was relatively small (n = 17), a significant improvement in fertilization (73.7%) and pregnancy (53.9%) rate was obtained when the testicular sample was preincubated for 24 h. This improvement prevailed in the obstructive azoospermic group, but was less pronounced in nonobstructive patients. CONCLUSIONS: This study shows that the outcome of fresh and frozen-thawed testicular spermatozoa in ICSI is comparable, obstructive and nonobstructive etiologies perform the same, and that preincubation of testicular spermatozoa results in increased fertilization and pregnancy rates. All testicular biopsies are therefore performed the day before oocyte retrieval, superfluous spermatozoa cryopreserved, and the remaining testicular homogenate preincubated for the 24 h prior to oocyte retrieval. With this regime, most azoospermic patients are treated successfully, irrespective of the use of fresh or frozen-thawed spermatozoa from obstructive or nonobstructive cases.

Cryopreservation↗
PubMed2001

Ultrastructural evaluation of recurrent and in-vitro maturation resistant metaphase I arrested oocytes.

An infertile couple whose female partner showed recurrent retrieval of immature metaphase I (MI) oocytes that were resistant to in-vitro maturation, was studied. Four spermiograms revealed teratozoospermia. Consistent non-fertilization and negative pregnancy outcomes were obtained after intrauterine insemination, gamete intra-Fallopian transfer and IVF. Two intracytoplasmic sperm injection (ICSI) cycles were finally performed. All oocytes (n = 17) in both cycles were arrested at MI and failed to mature after 48 h culture. ICSI also resulted in total non-fertilization. In the last cycle, two oocytes were analysed by transmission electron microscopy and showed almost identical results. All organelles showed normal characteristics of an MI oocyte. The main abnormality found was related to the MI spindle, with absence of microtubules and dispersion of the female chromosomes. Minor abnormalities were observed (immature fibrous appearance of the zona pellucida; the presence of small vesicle aggregates which formed a foam-like body). The injected sperm nucleus was arrested in the middle of the chromatin decondensation process, with no visible nuclear envelope reformation. Normal disruption of sperm acrosomal and flagellar components were observed. Only a partial cortical reaction was observed. This represents the first documented case of developmental arrest due to complete absence of spindle formation in association with an otherwise mature ooplasm.

Acrosome↗
PubMed1996

Fertilization and pregnancy using metaphase I oocytes in an intracytoplasmic sperm injection program.

PURPOSE: In this study we investigated whether metaphase I oocytes collected in an intracytoplasmic sperm injection program could successfully be matured and fertilized by injecting aged (> 20-hr) spermatozoa. MATERIALS AND METHODS: Metaphase I oocytes aspirated were preincubated for 20 hr to allow the oocytes to reach meiotic maturity. Only metaphase II oocytes were injected. The original sperm sample processed on the day of aspiration was used in the microinjection process. RESULTS: One hundred eighty-three oocytes were collected, of which 42 (23%) were metaphase I oocytes. These were incubated for 20 hr and microinjected with the original sperm sample. Thirty-one (74%) of the metaphase I oocytes reached meiotic maturity (extruded polar body): 67.7% showed two pronuclei 18 hr after injection and 61.3% embryo development 40 hr postinjection. No difference in fertilization and embryo development rate was found in metaphase II oocytes injected 6 hr postaspiration versus 20 hr postaspiration. An ongoing pregnancy was also achieved using only embryos obtained from matured metaphase I oocytes. CONCLUSIONS: Metaphase I oocytes can be successfully matured in vitro and injected using aged (> 20-hr) sperm samples. Matured metaphase I oocytes, if successfully injected, produce embryos able to induce pregnancy.

Embryo Transfer↗
PubMed1992

The influence of solubilized porcine zona pellucida protein on the binding capacity of human spermatozoa.

The acrosome reaction, sperm-zona pellucida binding, sperm-oolemma binding/fusion and subsequent fertilization are known to be influenced by homologous as well as heterologous follicular fluid and zona pellucida protein. In this study, the effect was investigated of different concentrations of solubilized porcine zona pellucida protein on the zona binding potential of human spermatozoa under hemizona assay conditions. Human spermatozoa incubated with 617 and 142 micrograms/ml porcine zona pellucida protein showed a statistically significant increase in zona binding when compared with control spermatozoa (106.5 +/- 18.0 versus 60.9 +/- 29.0, P less than 0.02 and 111.0 +/- 26.6 versus 63.0 +/- 25.5, P less than 0.0001, respectively). Concentrations of 67 micrograms/ml porcine zona pellucida protein did not show a significant increase in zona binding (78.7 +/- 21.7 versus 66.7 +/- 25.4, P greater than 0.05). Control zona binding values for different experiments did not differ significantly (60.9 +/- 29.0; 63.0 +/- 25.5; and 66.7 +/- 25.4, P greater than 0.6). In conclusion, it seems likely that a factor(s) present in the porcine zona pellucida might play a beneficial role during human sperm-oocyte binding. The results of the study might be used in future investigations to manipulate gamete interaction to such an extent that improved fertilization rates can be accomplished.

Animals↗
PubMed1991

Comparison of sperm binding potential of uninseminated, inseminated-unfertilized, and fertilized-noncleaved human oocytes under hemizona assay conditions.

In this study, human oocytes obtained after ovarian hyperstimulation for in vitro fertilization (IVF) and gamete intrafallopian transfer (GIFT) were utilized to evaluate sperm/zona pellucida binding potential. Three groups of oocytes were evaluated: 1) uninseminated; 2) inseminated-unfertilized; and 3) fertilized-uncleaved. All oocytes had undergone germinal vesicle breakdown at the time of retrieval and were salt-stored (pH 7.2) for not more than 30 days. Sperm binding was recorded under hemizona assay (HZA) conditions using spermatozoa from eight fertile men (HZA control) and from 1) four teratozoospermic (HZA test) and 2) four normozoospermic (HZA test) infertile men. First, the mean numbers (+/- SD) of sperm tightly bound for fertile controls and teratospermic men to hemizonae from uninseminated oocytes were 69.7 +/- 16 and 14.5 +/- 7, respectively (P = 0.02). Likewise, hemizonae from uninseminated oocytes bound 102.0 +/- 19 and 114.0 +/- 28, respectively, for fertile controls and normospermic men (P = 0.5). Second, hemizonae obtained from inseminated-unfertilized IVF oocytes bound 44.2 +/- 12 and 19.7 +/- 6 for fertile controls and teratospermic men, respectively (P = 0.02). This category of oocytes bound 100.5 +/- 7 and 108.5 +/- 11 sperm, respectively, for fertile controls and normospermic semen (P = 0.3). Third, HZA results of fertilized but uncleaved oocytes showed a mean number of tightly bound sperm of 6.0 +/- 4 compared with 65.0 +/- 1 in control, uninseminated oocytes using fertile sperm. These results demonstrate that uninseminated and inseminated-unfertilized human oocytes, salt-stored under controlled pH conditions, give reliable information regarding sperm binding potential under HZA conditions.

Female↗
PubMed1991

Detection of sperm antibodies on unwashed spermatozoa with the immunobead test: a comparison of results with the routine method and seminal plasma TAT titers and SCMC test.

This study investigated whether the immunobead test (IBT) could, for the purposes of simplicity and saving time, be applied directly on an unwashed semen sample instead of washed spermatozoa. These two methods were performed simultaneously on the semen samples of 15 men with a positive MAR test and 10 men with a negative MAR test. A possible association was found between the unwashed samples, showing positive IB binding (greater than 20%) on the tail and/or head and the seminal plasma TAT titers (P less than .00001, Fisher's exact test). In all cases with IB binding of greater than or equal to 20% on unwashed spermatozoa, positive seminal plasma TAT titers (greater than or equal to 32) and SCMC tests (greater than or equal to 50%) were found. In all cases where the binding of the beads was mainly located on the tailtips of the washed or unwashed spermatozoa, negative seminal plasma TAT titers and SCMC tests were found. Coating of the head and/or upper tail regions in both methods was always associated with high TAT titers and a strong-positive SCMC test. It is concluded that the IBT for IgA, but not for IgG, can be performed directly on unwashed semen and that the position of IB binding on the spermatozoa is of prognostic importance with regard to the expected outcome of the SCMC test and seminal plasma TAT titers.

Agglutination Tests↗
PubMed1990

The in vitro fertilisation programme at Tygerberg Hospital and the University of Stellenbosch. Five years' experience, April 1983-January 1988.

The results of the in vitro fertilisation programme at Tygerberg Hospital for the period April 1983 to January 1988 are presented. Of the 1117 laparoscopies performed, 825 patients reached the transfer stage. A live-birth rate of 9.3% was achieved. The pregnancy rate after transfer of 4 embryos was 25.9% compared with 15.4% after 2 embryos and 10.8% after 3 embryos (P = less than 0.0001). The multiple pregnancy rate was 2.8% in the group receiving 2 embryos and 11.7% and 10.4% in those receiving 3 and 4 embryos, respectively. Of the 77 successful pregnancies (90 babies), 1 baby died at 34 weeks' gestation as the result of abruptio placentae due to preeclampsia and 1 cot death occurred. The only congenital abnormality encountered was a cleft palate.

Female↗
PubMed1990

Treatment of male sperm autoimmunity by using the gamete intrafallopian transfer procedure with washed spermatozoa.

Sixteen couples were diagnosed as having immunological infertility. To detect sperm-bound immunoglobulin (Ig), i.e., IgA, IgG, and IgM antibodies, the direct immunobead test (IBT) was used. In each individual patient, the direct IBT was greater than or equal to 70% positive for either IgA or IgG or both. The indirect IBT was positive for IgA and IgG antibodies in the serum of all the patients. Semen was collected in 15 mL medium (Ham's F10 [Gibco, Grand Island, NY] + 10% whole blood serum) and prepared with the wash and swim-up method. Patients in the study group were treated for their immunological infertility problem by performing the gamete intrafallopian transfer (GIFT) procedure. An ongoing pregnancy was achieved in 7 of the 16 (43%) couples treated with the GIFT procedure with an ongoing pregnancy rate of 24.1% (7 of 29) per cycle. The GIFT procedure appears to be an effective and safe way of treating male immunological infertility.

Autoantibodies↗
PubMed1989

Use of specific monoclonal antibodies to secretory IgA for the detection of spermatozoal antibodies in serum and seminal plasma by enzyme-linked immunosorbent assay.

The role of anti-sperm secretory IgA has recently received attention since some workers feel it plays an important role in the prognosis of the immunologically infertile couple. Current methods used in our laboratory cannot separately detect anti-sperm secretory IgA. An enzyme-linked immunosorbent assay (ELISA) utilizing specific monoclonal antibodies to secretory IgA was used to detect anti-sperm secretory IgA as well as anti-sperm monomeric IgA and IgG in serum and seminal plasma of a sperm-antibody-positive (ASA+) and sperm-antibody-negative (ASA-) group of men. Results showed significantly raised serum levels in the ASA+ group when compared to the ASA- group for anti-sperm secretory IgA (P less than .001), anti-sperm monomeric IgA (P less than .001), and anti-sperm IgG (P less than .01). Seminal plasma levels were also raised in the ASA+ group, but only significantly so for monomeric IgA (P less than .02). The performed ELISA has definite potential in research, especially with the use of monoclonal antibodies for the detection of anti-sperm secretory IgA, but cannot as yet be used as a prognostic predictor of fertility in the individual antibody-positive patient. Infertility specific antigens will have to be identified and isolated and subsequently used in the ELISA.

Antibodies, Anti-Idiotypic↗
PubMed1989

Effect of sperm washing and swim-up on antibodies bound to sperm membrane: use of immunobead/sperm cervical mucus contact tests.

The direct immunobead test (IBT) and the sperm cervical mucus contact (SCMC) test were used to evaluate the effect of sperm washes and swim-up on antibodies bound to the sperm membrane in 11 patients with autosperm antibodies (30-100% IgA and 30-100% IgG) but otherwise normal semen measurements. The tests were performed on semen samples before and after a wash/swim-up procedure in Ham F10 + 10% human blood serum. Sperm-bound antibodies in washed spermatozoa that were able to swim into a layer of medium did not differ from those in unwashed samples. The IBT and the SCMC test on unwashed and washed spermatozoa were also not significantly different. However, all other washed semen parameters were improved. Correct washing of semen samples is important when performing the IBT to prevent false negative results. Eight of the 11 partners (73%) of the patients tested became pregnant after treatment with washed sperm. The ongoing pregnancy rate was 64%.

Animals↗
PubMed1989

Effect of rapid dilution of semen on sperm-bound autoantibodies.

The effect of rapid dilution of autoantibody-positive semen on sperm-bound antibodies was studied in 12 male patients (0-100% IgA and 10-100% IgG). The direct immunobead test (IBT) and the sperm cervical mucus contact (SCMC) test were used to detect sperm-bound antibodies on spermatozoa before and after rapid dilution and swim up in HAM F-10 + 10% human blood serum. All patients tested had normal semen parameters. Sperm-bound antibodies detected after swim-up of semen samples ejaculated into 15 ml HAM F-10 + 10% serum (diluted) did not differ significantly from undiluted samples. Most values were lower (IBT and SCMC) but were not statistically significant. Sperm motility, forward progression, and morphology were statistically improved. Pregnancies resulting from washed semen (diluted) in combination with artificial insemination (AIH) or gamete intrafallopian tube transfer (GIFT) took place in spite of antibodies still present on the sperm membrane and can probably be attributed to improved semen quality, minimizing of ovum-sperm distance, and cervical mucus elimination.

Autoantibodies↗
PubMed1989

Antisperm antibody tests: traditional methods compared to ELISA.

Enzyme-linked immunosorbent assays (ELISAs) to detect sperm antibodies can be a useful addition to other tests. ELISAs allow more quantitative and detailed information than some other tests, but results should be compared to those of other tests. This study was conducted to correlate results of an ELISA that used a sperm membrane extract as antigen with those of other tests. Semen, seminal plasma, and serum of 34 sperm-antibody-positive and 36 sperm-antibody-negative men were tested. There was poor correlation between the ELISA results and those of the other tests. Only the results of a direct immunobead test done on semen correlated fairly well with results from ELISAs done on serum. Further studies are needed on fertility-specific antigens that can be identified and isolated and then studied by means of ELISAs.

Antibodies↗
PubMed1988

Evaluation of immunobead test (IBT), tray-agglutination test (TAT), and sperm immobilization test (SIT).

The present investigation relates the results obtained by the indirect immunobead test (IBT) to those found during the tray-agglutination test (TAT) and sperm immobilization test (SIT) performed on 16 positive blood sera samples. An IB binding of greater than 50% can be regarded as significantly positive (p less than 0.05), since 77% of the positive IB samples with an IB binding of greater than 50% also revealed a TAT titer of greater than 1:8. The IBT is an excellent test for the detection of sperm antibodies during routine screening procedures.

Agglutination Tests↗
PubMed1982

The effect of paraquat on the incorporation of radiolabelled proline into acid-extractable lung proteins and collagens.

In this comparative study we describe the influence of paraquat on the rate of L-2,3-3H-proline incorporation into the acid-extractable proteins and into newly synthesized protropocollagen molecules from the lungs of rats and rabbits. Exposure to paraquat took two forms: (a) addition of paraquat in vitro to lung tissue taken from rats and rabbits, and (b) intraperitoneal injection of paraquat prior to death. Paraquat (0,5 - 1,0 mM) added in vitro significantly slowed the rates of 3H-proline incorporation into the acid-extractable proteins and into newly synthesized protropocollagen in both rat and rabbit lung tissue. Paraquat administered intraperitoneally (27 mg/kg) to rabbits did not markedly influence the rate of 3H-proline incorporation into acid-extractable proteins and collagen assessed in vitro 24, 48 and 96 hours after injection. Paraquat injected intraperitoneally into rats induced no significant difference in synthesis rates of acid-soluble proteins up to 48 hours after injection. During the same period, the collagen synthesis rate of rat lung tissue was reduced. At 96 hours an increase was found when the rate of synthesis was expressed as cpm/mg DNA and as cpm/microgram hydroxyproline.

Animals↗