PubMed Health⌕ Search

Biomedical subjects

D R Franken

Publications and source records attributed to D R Franken.

At least 19 recordsLinked to original sources
PubMed1995

Fluorescein isothiocyanate conjugate-wheat germ agglutinin staining of human spermatozoa and fertilization in vitro.

OBJECTIVE: To determine the sensitivity and clinical significance of fluorescein isothiocyanate wheat germ agglutinin (FITC-WGA) staining of human spermatozoa. DESIGN: Percentage FITC-WGA staining was assessed in the different morphological zones of human spermatozoa and compared with fertilization rates of metaphase II oocytes in vitro. PATIENTS: Spermatozoa and oocytes were obtained from normospermic (n = 12) and male factor patients (n = 15) attending the Tygerberg Hospital's IVF clinic. MAIN OUTCOME MEASURES: Aliquots of swim-up spermatozoa used in the IVF laboratory were examined for the presence and morphological localization of WGA receptors. Fertilization rates were expressed as the percentage of metaphase II oocytes with successful fertilization. RESULTS: Human spermatozoa capacitated in vitro for use in an IVF program showed variations in the percent positive staining of the acrosomal region which correlated with IVF rates in both patient groups studied. Values of FITC-WGA acrosomal staining of < 35% corresponded with IVF rates of < 50% whereas > 35% WGA acrosomal staining corresponded with IVF rates > or = 50%. Significant differences between acrosomal and equatorial region staining from normozoospermic samples and male factor samples also were observed. CONCLUSION: The results indicate the practical use of the WGA assay for molecular assessment of male fertilizing capacity in conjunction with an assisted reproductive program.

Acrosome↗
PubMed1994

Selective capacity of glass-wool filtration for the separation of human spermatozoa with condensed chromatin: a possible therapeutic modality for male-factor cases?

PURPOSE: The aim of this study was to evaluate chromatin condensation of human spermatozoa following swim-up compared to glass-wool separation. Semen aliquots from men attending an andrological outpatient clinic were processed by means of a swim-up procedure and glass-wool filtration. Chromatin condensation was recorded using aniline blue staining and results were reported according to color intensity of stained sperm heads. Morphometric measurements of sperm heads were performed on stained sperm samples. RESULTS: Glass-wool filtration resulted (i) in a significantly higher total motile sperm count (P < 0.0005) compared to swim-up and (ii) in a significantly higher percentage of normal chromatin-condensed spermatozoa compared to the ejaculate (P < 0.01). CONCLUSION: In contrast, comparing swim-up to the ejaculate, the percentage of matured nuclei (unstained spermatozoa) retrieved following swim-up was significantly lower (P < 0.005). Glass-wool filtration separates human spermatozoa according to motility and size of the sperm head. The size of the sperm head closely correlated with the chromatin condensation quality.

Aniline Compounds↗
PubMed1992

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↗
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

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↗
PubMed1991

Hemizona assay: use of fresh versus salt-stored human oocytes to evaluate sperm binding potential to the zona pellucida.

The hemizona assay (HZA) has a high predictive value for in vitro fertilization (IVF) results. Oocyte quality plays a significant role in the validation of this test. The question was asked whether human salt-stored oocytes (up to 30 days) are damaged and subsequently lose their sperm binding capacity when compared to fresh human oocytes. Equivalent binding in both the salt-stored and the fresh group of oocytes was observed in the hemizonae incubated with normal semen as well as in their matching halves incubated with semen from an infertile man. Based on the results, we conclude that salt-stored oocytes (pH 7.0) give reliable information regarding sperm binding potential under HZA conditions.

Female↗
PubMed1991

Defining the valid hemizona assay: accounting for binding variability within zonae pellucidae and within semen samples from fertile males.

OBJECTIVE: To achieve a better understanding of the variability in sperm and oocyte binding capacities will optimize use of the hemizona assay (HZA) as a predictor of sperm function. DESIGN: Limitations of the HZA were more clearly delineated by current studies: (1) variability of sperm binding capacity of men over a 90-day interval; (2) variability of sperm binding using different oocytes; and (3) lower limits of the number of sperm bound from the fertile control in two laboratories. PATIENTS: Semen was obtained from proven fertile men and one subfertile individual. MAIN OUTCOME MEASURE: The number of sperm tightly bound to the hemizona were measured and compared. RESULTS: In the initial study, 6 fertile control men exhibited a similar degree of variability in zona binding when studied over a 90-day interval. Average sperm binding for individuals ranged from 68 to 127. Second, 3 of the 15 simultaneous assays showed very low numbers of sperm bound, indicating that 20% of the zonae had poor binding. Third, from 18 men who had 0% fertilization in an in vitro fertilization system using mature oocytes, evaluation of their sperm by HZA was performed. The sperm bound poorly and the 95% confidence interval was 20 sperm bound. Thus, the fertile controls should bind greater than 20 sperm to distinguish them from the infertile group in the HZA system resulting in a valid assay. CONCLUSIONS: With these guidelines, applications of the HZA may be made with greater reassurance of a valid bioassay of sperm fertilizing potential.

Biological Assay↗
PubMed1990

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↗
PubMed1989

Hemizona assay using salt-stored human oocytes: evaluation of zona pellucida capacity for binding human spermatozoa.

Human oocytes were stored (25 degrees C) in 1.5 M MgCl2 for 6-30 days, then utilized in the new hemizona assay (HZA) for tight binding of human spermatozoa [Burkman et al.: Fertil Steril 49:688-697, 1988]. We have compared 1) the ability of matching salt-treated hemizonae or dimethylsulfoxide (DMSO)-treated hemizonae to distinguish between sperm from semen having normal versus subnormal characteristics and, 2) the kinetics of fertile sperm binding to salt-treated or DMSO-treated hemizonae. After sperm preparation one salt-treated hemizona was incubated with normal spermatozoa and the matching hemizona was placed with sperm from the subnormal group. As a control, DMSO-treated hemizonae were incubated in additional sperm droplets. After 4 hours, the number of sperm tightly bound to each hemizona was counted. Within the normal semen group, there was equivalent binding to salt- or DMSO-treated hemizonae (54.0 +/- 12 and 49 +/- 14, respectively, mean +/- SEM). Similarly, tight binding of sperm from the subnormal group was not affected by the zona storage method (21 +/- 8 and 17 +/- 5, respectively). For either storage approach, binding of subnormal sperm was significantly less (P less than 0.01) compared with the number of normal sperm attached to the matching hemizona. For the kinetics study, the hemizona binding of proven fertile spermatozoa was followed throughout 8.5 hours. The shape of the binding curve was the same for zonae stored by either method and was consistent with our published kinetics data. Salt storage offers a simple and inexpensive means for accumulating and transporting human zonae pellucida; the resulting hemizonae function effectively in the HZA for estimating sperm binding potential.

Dimethyl Sulfoxide↗
PubMed1989

The hemizona assay (HZA): a predictor of human sperm fertilizing potential in in vitro fertilization (IVF) treatment.

The hemizona assay (HZA) was developed to assess human sperm fertilizing potential. This blinded study investigated the relationship between sperm binding to the hemizona and in vitro fertilization (IVF) success (36 patients). Nonliving human oocytes were recovered from excised ovaries and stored. Each zona pellucida was cut into equal hemispheres by micromanipulation. For the HZA, one droplet exposed a hemizona to abnormal spermatozoa, while the control droplet contained the matching hemizona and spermatozoa from normal semen. After 4 hr, the number of tightly bound spermatozoa was counted. Binding to the hemizona was significantly higher for those having IVF success (mean of 36.1 +/- 7, versus 10.4 +/- 4 from the failure group; P less than 0.05). Fewer sperm from the failure group had a strictly normal morphology (3.2 versus 12.7%; P less than 0.05, Kruger method). Tight zona binding was significantly correlated with the percentage motile sperm, percentage normal morphology, and seminal sperm concentration. These results enhanced our confidence that the HZA is diagnostic for identification of patients at high risk of failing to achieve fertilization in vitro.

Female↗
PubMed1989

Murine oocyte cryopreservation: comparison between fertilization success rates of fresh and frozen metaphase I and II oocytes.

The surplus of oocytes obtained during in vitro and gamete intrafallopian transfer creates the need to either (1) cryopreserve (196 degrees C) or (2) fertilize these surplus oocytes and then freeze the resulting embryos. During this pilot study, C57BL6 X CBA female mice (F1 generation) were superovulated by means of hormone stimulation, resulting in adequate numbers of murine oocytes. Oocytes thus obtained were classified according to their nuclear status. The maturation stage of oocytes was evaluated and classified as metaphase I and metaphase II. The success of oocyte cryopreservation in each group was evaluated according to fertilization rate after the insemination procedure. Metaphase II oocytes had a fertilization rate of 33% (frozen) and 84% (fresh), whereas metaphase I oocytes had a 24% (frozen) and a 79% (fresh) fertilization rate. Metaphase I oocytes can mature to the metaphase II stage after having been frozen, metaphase II oocytes may be more vulnerable to chromosome damage, and the nuclear stage prior to freezing has no significant effect on the fertilizability of the frozen-thawed oocytes.

Animals↗
PubMed1989

Validation of the hemizona assay in a monkey model: influence of oocyte maturational stages.

The hemizona assay (HZA) was used in the monkey model to investigate sperm binding to the zona pellucida and to evaluate binding patterns according to various stages of oocyte development. Adult cynomolgus monkeys were superovulated with human menopausal gonadotropin. The oocytes were retrieved by laparoscopy, stored in salt solution, then cut into hemizonae by micromanipulation. Metaphase II oocytes showed significantly tighter binding than prophase I oocytes (P less than 0.001). Metaphase I oocytes showed intermediate binding, significantly different from the other groups (P less than 0.01). It is concluded that: (1) This study demonstrates the feasibility of HZA as a test to evaluate sperm/zona interactions in the monkey, using standards reported for human studies. (2) Oocyte meiotic competence seems to be accompanied by an increase in zona-binding properties.

Animals↗
PubMed1988

The SCMC test: a reliable monitor for antispermatozoal antibodies.

Seventeen couples (13%) were selected from a group of 129 infertile patients according to the following criteria: (i) unexplained infertility for 3 years and (ii) greater than 50% shaking spermatozoa during SCMC testing. The couples were tested for sperm antibodies after a complete diagnostic work-up schedule. Post-coital tests were performed during the first menstrual cycle of the wife, followed by SCMC and sperm antibody titre testing. Ten males and seven females were thus treated with 96 mg methylprednisolone. Nine (52%) of the 17 with sperm antibodies achieved a pregnancy. The results of the SCMC test were in all the cases indicative of the actual sperm antibody titre. Reduction of the antibody titre and a decrease in the percentage of shaking spermatozoa as detected by the SCMC test correlated well with the pregnancy rate amongst the patients.

Antibodies↗
PubMed1988

The hemizona assay (HZA): development of a diagnostic test for the binding of human spermatozoa to the human hemizona pellucida to predict fertilization potential.

The authors present their initial results with the hemizona assay (HZA), which was developed to predict the fertilizing potential of spermatozoa. The HZA uses the matching halves of a human zona pellucida from a nonfertilizable and nonliving oocyte, providing an internal control on zona-to-zona variability. Maximal binding of human sperm to the hemizona usually occurred after 4 to 5 hours of coincubation. Sperm from fertile men exhibited significantly higher binding capacity to hemizonae compared with sperm from men who had fertilization failure during in vitro fertilization (IVF) treatment. The HZA index is calculated as follows: (bound sperm from subfertile male) divided by (bound sperm from fertile male) X 100. These findings demonstrate that the HZA may be a useful diagnostic tool in male infertility evaluations.

Female↗
PubMed1987

The immunobead test as a sperm antibody detector.

The occurrence of sperm antibodies on the surface of sperm can be demonstrated by several methods. A study to evaluate the immunobead (IB) test as a laboratory procedure during the detection of sperm antibodies among infertile males showed an overall prevalence of 6.8% and 5.8% for IgG and IgA respectively--convincing evidence that the IB test can be employed as a sperm antibody detector.

Autoantibodies↗
PubMed1987

Gamete intrafallopian transfer: a treatment for long-standing infertility.

Gamete intrafallopian transfer (GIFT) has been described by Asch et al. (1,2) as an alternative technique in the treatment of infertile couples. At the University of the Orange Free State, the GIFT technique was introduced in July 1985, and during phase I, 31 patients were treated by means of GIFT. All patients had had at least six cycles of ovulation induction with human menopausal gonadotropin (hMG) and human chorionic gonadotropin (hCG). Their diagnoses were anovulation (3 patients), mild endometriosis (17 patients), and unexplained infertility (11 patients). All husbands' semen analyses were normal. Of the 31 patients, 3 failed, due to spontaneous ovulation before laparoscopy (1 patient) and unsuccessful follicle aspiration at laparoscopy with no oocytes found (2 patients). Four ongoing pregnancies resulted from the remaining 28 patients. This represents a pregnancy rate of 14.29% per laparoscopy (including the failures). The patients who became pregnant had had infertility treatment for 5, 6, 8, and 8 years, respectively. GIFT therefore appears to be a promising method of treatment for long-standing infertility.

Adult↗