Sperm morphology--predictive value?
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Animals↗
Biomedical subjects
R Menkveld
Publications and source records attributed to R Menkveld.
The Acridine Orange test: determining the relationship between sperm morphology and fertilization in vitro.
The prediction of human fertilization is an important aspect of research protocols dealing with male fertility. Sperm DNA has been reported to be an indicator of human sperm fertility potential. The Acridine Orange test for evaluation of sperm DNA has been employed during the present study to determine its relationship with human sperm morphology and fertilization in vitro. Seventy-six patients from the in-vitro fertilization (IVF) and/or gamete intra-Fallopian transfer (GIFT) programme were randomly selected for the study. All patients had a routine semen analysis, sperm DNA evaluation and underwent standard IVF procedures at the time of the study. The results indicated a moderate positive correlation (r = 0.38, P = 0.0006) between results of the Acridine Orange test and normal sperm morphology. Patients with an Acridine Orange test value exceeding 24% had significantly higher oocyte fertilization rates than patients with lower values, for metaphase I (74 versus 51%, P = 0.0008) and for metaphase II oocytes (88 versus 60%, P = 0.0001). Sperm morphology, however, proved to be a more significant predictor of fertilization in vitro compared to the Acridine Orange test.
Acridine Orange↗
Sperm selection capacity of the human zona pellucida.
Previous hemizona assay (HZA) results have illustrated a positive and significant correlation between the percentage of morphologically normal spermatozoa in the semen and the number of spermatozoa tightly bound to the zona pellucida. The present study was designed to evaluate the morphologic features using strict criteria of spermatozoa tightly bound to the zona pellucida. Semen samples of 4 normozoospermic and 11 teratozoospermic men were used to compare the percentage of normal spermatozoa in the semen with that found 1) after swim-up separation and 2) bound to the zona under HZA conditions. The mean (+/- SEM) % normal forms for normozoospermic men in semen, after swim-up and zona-bound spermatozoa were 21.5 +/- 1.6, 27.5 +/- 2.9, and 44.8 +/- 3.4, respectively. A significantly higher % of normal forms were found among zona-bound sperm compared to swim-up forms (p = 0.02) and seminal sperm (p = 0.02). The mean % of normal sperm forms present in semen, after swim-up and zona pellucida-binding for teratozoospermic men, were 3.7 +/- 0.9, 5.8 +/- 1.6 and 15.6 +/- 3.1, respectively. Significant differences existed between the % of normal sperm forms found in the swim-up and zona-bound spermatozoa (P = less than 0.01 and P = less than 0.0003, respectively) compared to the original ejaculates. Results indicate that a selective process against abnormal spermatozoa occurs at the site of the zona pellucida.(ABSTRACT TRUNCATED AT 250 WORDS)
Cell Separation↗
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↗
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↗
Hemizona assay and teratozoospermia: increasing sperm insemination concentrations to enhance zona pellucida binding.
This study aimed: (1) to evaluate the zona-binding capacity of patients with abnormal sperm morphology, using standard hemizona assay (HZA) conditions and increasing sperm insemination concentration during the assay and (2) to determine the insemination concentration needed to obtain equality in the number of tightly bound sperm to matching hemizonae, using sperm from teratozoospermic patients versus proven fertile controls. The minimum concentration of motile sperm from fertile controls necessary to validate HZA results was 250,000/mL (35.4 +/- 5.6 tightly bound sperm; mean +/- SE). The "effective number of sperm" (morphologically normal with high motility) was 60,750/mL. Each teratozoospermic patient had a unique, (higher) sperm insemination concentration (range: 0.5 X 10(6) to 2.0 X 10(6) motile sperm/mL) necessary to equal the number of tightly bound sperm representing the lower 95% confidence interval for the control sample (at 0.5 X 10(6) motile sperm/mL) with the matching hemizona. These results suggest that the HZA may be used as an indicator of the sperm insemination concentration during in vitro fertilization in patients with teratozoospermia.
Female↗
The evaluation of morphological characteristics of human spermatozoa according to stricter criteria.
The evaluation of the morphology of human spermatozoa varies widely between and sometimes even within laboratories. The purpose of this study was to determine whether the method that has been developed in our laboratory and which resulted in the use of stricter criteria for the evaluation of sperm morphology is a practical, reliable and repeatable method and to establish the within and between observer variations. The criteria used for a 'normal' spermatozoon are based on the appearance of spermatozoa found in the mucus of the upper endocervical canal. The results of the morphological evaluations of 26 samples by four observers were statistically analysed by various methods. The method of Barnett showed a high degree of relative accuracy between observers with error variances of between 2.89 and 19.67 as well as high Spearman rank correlation coefficients of between 0.8675 and 0.6537 (P less than 0.0003). The Spearman correlation coefficient for 15 duplicate evaluations by one observer was 0.9650 (P less than 0.0001) while the coefficients of variation for repeated evaluations of single samples were also within acceptable limits. Based on these results, the method described in this article allows comparable and reliable results between and within observers to be obtained. From this and other studies it can be concluded that the method also has a good prognostic value for the prediction of expected IVF fertilization, the hamster test and hemizona assay.
Cytodiagnosis↗
Human pregnancy after transfer of intact frozen-thawed embryos.
Since the birth of the first baby as a result of in vitro fertilisation (IVF) in 1978, many clinics around the world have achieved pregnancies and births for their patients by using IVF and gamete intrafallopian transfer procedures. With the storage of excess embryos, multiple laparoscopies can be avoided; this has favoured the development of better cryopreservation techniques. In our clinic 8-cell human embryos are frozen in a 1.5M dimethyl sulphoxide solution as cryoprotectant using the slow freeze-thaw method. Sixteen thawed embryos were replaced in 8 patients, resulting in 1 pregnancy. Of the thawed embryos 51.6% survived the freezing process in that they had 50% or more of the original number of blastomeres and also the zona pellucida intact.
Cryopreservation↗
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↗
Semen characteristics of vervet monkeys.
Semen samples (91) from 47 vervet monkeys were collected by electroejaculation over a 2 year period. Seventy-eight of these were from 37 singly caged males of unknown fertility and 13 from 10 breeding males of known fertility. Mean values for semen characteristics of the singly caged males were: volume 0.45 ml, pH 7.8, concentration 184 x 10(6)/ml, forward progression rating 2.95 (scale 0-4), motility 55.4%, live 68% and abnormal morphology 3.5%. Mean values for semen characteristics for the breeding males were: volume 0.86 ml, pH 9.00, concentration 117.15 x 10(6)/ml, forward progression rating 3.00 (scale 0-4), motility 43.6%, live 53.3% and abnormal morphology 6%. Semen volumes in the singly caged males were lower than the volumes reported in other studies.
Animals↗
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↗
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↗
Does the wash-up and swim-up method of semen preparation play a role in sex selection?
Because of the higher built ratio of boys to girls (1.5 : 1) by in vitro fertilization (IVF) and gamete intrafallopian transfer (GIFT) at Tygerberg Hospital, the question arose as to whether the wash-up and swim-up method of semen preparation, used for our IVF procedure, plays a role in sex selection. Semen samples of 20 men were evaluated at different time intervals to determine the percentage of F-body-positive spermatozoa. It was found that the swim-up time affects the relationship between Y- and X-bearing spermatozoa, especially at time intervals of 30 and 45 min, where an optimum percentage of Y-bearing spermatozoa (p = 0.0039 and p = 0.0092, respectively) was found. Sperm obtained at these specific time intervals may influence the outcome of sex by IVF and GIFT in favor of males.
Fertilization in Vitro↗
Critical assessment of sperm morphology before and after double wash swim-up preparation for in vitro fertilization.
Critical assessment of sperm morphology using specific and stringent criteria is predictive of the subsequent ability of those sperm to fertilize oocytes in in vitro fertilization (IVF). Previous studies have evaluated sperm morphology prior to sperm preparation and, thus, have not assessed the actual sperm used for insemination. We studied the impact of a double wash swim-up technique used for IVF on sperm morphology using the strict criteria of Kruger et al. [5, 6] in 73 consecutive patients undergoing IVF. Pre- and postswim-up morphological assessments were done in a prospective, randomized, and blinded fashion. The mean percentage of normal forms pre- and postswim-up was 19.8% and 23.4%, respectively, an improvement of 18% (p less than 0.05) with 62 of 73 patients showing improvement. Significantly, analysis of the 27 patients with abnormal morphology on initial assessment (normal forms less than or equal to 14%) revealed an improvement in percentage of normal forms from 9.0% to 21.5%, a mean increase of 239% (p less than 0.005). Eighteen of these 27 patients showed improvement in their percentage of normal forms, eight were unchanged, and only one patient had a decline in their percentage of normal forms. We conclude that the double wash swim-up preparation used for IVF substantially improves the percentage of sperm with normal morphology, and that the benefit is most substantial in those cases where the percentage of normal morphology is abnormal.
Fertilization in Vitro↗
Hypoosmotic swelling test in the prediction of male fertility.
The hypoosmotic swelling test (HOS) is a simple test to measure the functional competence of human sperm membranes. The question is, does a relationship exist between this competence and the fertilization potential of human spermatozoa? In this study the strongest correlation (r = 0.76) was obtained between sperm swelling and sperm viability (supravital staining). Only a moderate correlation (r = 0.50) was obtained with normal sperm morphology; weaker correlations were also obtained with the sperm penetration assay (r = 0.42) and human IVF (r = 0.24). The results, therefore, indicate that the HOS test has a limited predictive value. Notwithstanding this low concordance between sperm swelling and fertilizing potential, a less than 50% HOS test threshold was seen to be a definite indicator of a male factor.
Cell Survival↗
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↗
Usefulness of sperm penetration assay in fertility predictions.
The competence of the sperm penetration assay (SPA) to predict male fertility, as determined by normal sperm morphology and the fertilizing potential, as shown by human in vitro fertilization (IVF), was investigated. A significant correlation was obtained between normal sperm morphology and the SPA (phi = 0.623). A weaker correlation was however obtained with human IVF (phi = 0.397). Notwithstanding this weak association, a positive SPA (greater than 10%) was highly predictive (95%) of human IVF success. In contrast, a negative SPA (less than or equal to 10%) was associated with a high rate of false-negatives (65%). The SPA does however warn that a male factor may be present, as the mean fertilization rate of this group of patients was markedly reduced. The preincubation period for the spermatozoa did not play a major role in the predictive ability of a SPA outcome.
Female↗
Sperm morphologic features as a prognostic factor in in vitro fertilization.
To determine whether there is a prognostic value in the percentage normal sperm morphologic features in a human in vitro fertilization (IVF) program, the authors conducted a prospective study in women with bilateral tubal damage. Based on the percentage of morphologically normal spermatozoa, the patients were divided into four groups: group I, normal morphologic features between 0% and 14%; group II, 15% to 30%; group III, 31% to 45%; and group IV, 46% to 60%. One hundred ninety successful laparoscopic cycles were evaluated. In group I, 104 oocytes were obtained, of which 37% fertilized, but no pregnancy resulted; in group II, 81% of 324 oocytes were fertilized, with a pregnancy rate per embryo transfer (ET) of 22%; in group III, 82% of 309 oocytes were fertilized, with a 31% pregnancy rate; and in group IV, 91% of 69 oocytes were fertilized, with a pregnancy rate of 12%. Probability models indicated that there was a clear threshold in normal sperm morphologic features at 14%, with high fertilization and pregnancy rate in the groups with normal sperm morphologic features greater than 14%.
Embryo Transfer↗