Biomedical subjects

D R Franken

Publications and source records attributed to D R Franken.

At least 37 records · Page 2Linked to original sources

PubMed1999

Use of failed-fertilized oocytes for diagnostic zona binding purposes after sperm binding improvement with a modified medium.

PURPOSE: Because the availability of prophase oocytes for zona binding testing is limited, we compared sperm binding to the zona of failed-fertilized intracytoplasmic sperm injection (ICSI) and in vitro fertilization (IVF) oocytes after incubation in a standard IVF medium and a specially composed binding improvement medium. METHODS: Semen samples from nine patients and nine fertile donors were separated in parallel by the standard swim-up method in both media. Subsequently, hemizona assays were performed with prophase, failed-fertilized ICSI and IVF oocytes. RESULTS: Sperm separation resulted in a significantly higher sperm count (P < 0.01) and progressive motility (P = 0.018) in binding improvement medium. Moreover, spermatozoa coincubated with hemizonae (prophase, failed-fertilized ICSI and IVF oocytes) in binding improvement medium bound significantly more to hemizonae than in the controls (P < 0.01). However, the hemizona index did not differ. CONCLUSIONS: Thus, the limited number of human zonae can be increased by the use of oocytes that failed to fertilize during ICSI or IVF. This will lead to a qualitative improvement of the diagnostic spectrum in male-factor infertility.

Adolescent↗

PubMed1999

Normal sperm morphology and chromatin packaging: comparison between aniline blue and chromomycin A3 staining.

The successful implementation of ICSI has provided a unique means of allowing couples suffering from severe male infertility to achieve their reproductive goals. However, despite the great therapeutic advantages of the technique, ICSI often provides solutions to clinicians in the absence of an aetiological or pathophysiological diagnosis. The development of a sequential diagnostic schedule for patients consulting for fertility disturbances would be an ideal method of approach. Since sperm morphology recorded by strict criteria has often been correlated with fertilization failure, the present study aimed to evaluate the relationship between normal morphology and chromatin staining among fertile and subfertile men. Both chromomycin A3 (CMA3) and acidic aniline blue (AAB) were employed to record chromatin packaging quality among 58 men visiting the andrology laboratory. Intra- and interassay variations were initially recorded for fertile sperm donors. The coefficients of variation (CV) for all intra- and inter-assay assessments were < 12%. Chromatin packaging was significantly and negatively correlated with normal sperm morphology, namely r = 0.40 (P = 0.001) and r = 0.33 (P = 0.001) for CMA3 and AAB, respectively. Receiver operator characteristics illustrated sensitivity and specificity values of 75% and 82% for CMA3 and 60% and 91% for AAB, respectively. Significantly different CMA3 and AAB staining was recorded among men with severe teratozoospermia (< 4% normal forms) when compared with normozoospermic men (> 14% normal forms), namely 49% vs. 29% for CMA3 and 51% vs. 26% for AAB staining, respectively. Chromatin packaging assessments should be a valuable addition to the sequential diagnostic programme in an assisted reproduction arena.

Aniline Compounds↗

PubMed1999

Characterization of human zona pellucida glycoproteins.

The human egg may only be fertilized by one spermatozoon to prevent polyploidy. In most mammals, the primary block to polyspermy occurs at the zona pellucida (ZP). Little is known of the human ZP and the changes occurring following fertilization to prevent polyploidy. Using antibodies directed against synthetic peptides predicted from the human ZP2 and ZP3 cDNA, we identified ZP3 as a 53-60 kDa glycoprotein and ZP2 as a 90-110 kDa glycoprotein in prophase-I oocytes. Characterization of the ZP from metaphase II arrested eggs (inseminated-unfertilized and fertilized-uncleaved), shows no visible modification of ZP3, but demonstrates that ZP2 undergoes limited proteolysis in the amino terminal domain, to a 60-73 kDa species, denoted ZP2p, which remains linked to the proteolysed fragments by intramolecular disulphide bonds. A lack of ZP2 proteolytic activity in acrosomal supernatants is consistent with an oocyte origin for the protease. The ZP2-specific protease may be released during cortical granule exocytosis which occurs during meiotic maturation and following sperm-egg fusion as part of the block to polyspermy. Since mouse ZP2 acts as a secondary sperm receptor, it is possible that intact ZP2 binds a secondary egg binding protein, whereas cleaved ZP2 does not, suggesting a possible mechanism for the block to polyspermy.

Acrosome Reaction↗

PubMed1998

The clinical significance of sperm-zona pellucida binding.

The development of homologous functional bio-assay for sperm quality assessment has been a focal point of reproductive biologists; in order to provide a scientific based diagnosis in cases of fertilization failure. The availability of oocytes still remains an important limiting factor for laboratories to embark on the methodology of the assay. The use of zonae pellucidae, derived from post mortem and different in vitro fertilization oocytes, enhanced to availability of zonae. Sperm-zona binding has been illustrated to be an essential requisite during human fertilization and can be measured under hemizona assay as well intact zona pellucida conditions. The sensitivity and specificity of sperm-zona binding results indicated the assay to be positively and significantly correlated with in vitro fertilization outcome. Furthermore, a highly significant correlation were illustrated to exist between the normal sperm morphology, hyperactivation, sperm creatine kinase activity and the zona binding capacity of a given sperm sample. It was concluded that andrology testing remains an ever-growing component in the work-up of the infertile couple. We enter the next millennium with many questions that remain to be answered by the hand of efficacious screening techniques and a new formidable therapy in intra cellular sperm injection.

Female↗

PubMed1996

Defining bioassay conditions to evaluate sperm/zona interaction: inhibition of zona binding mediated by solubilized human zona pellucida.

OBJECTIVE: Our goal was to study the influence of solubilized human zonae pellucidae on zona binding potential and acrosome reaction. MATERIALS AND METHODS: Zona pellucida (ZP) solutions were prepared by dissolving zona in acidic buffer, NaH2PO4 (pH 2.5), to obtain 0.1 and 0.5 zona pellucida/microliters. Zona binding capacity was evaluated by the addition of oocytes (10-fold) to sperm/zona pellucida solution droplets. The number of sperm bound to each oocyte was recorded. Zona pellucida-mediated acrosome activity was evaluated after 60 min of coincubation of sperm and 0.5 ZP/microliters. RESULTS: The mean (+/- SE) number of sperm bound for control, 0.1 ZP/microliter, and 0.5 ZP/microliter was 181.2 +/- 12, 79.6 +/- 5, and 38.8 +/- 3, respectively. Zona pellucida-exposed sperm populations showed significant more acrosome-reacted sperm compared to control sperm, namely, 78 versus 32%, respectively (P = 0.001). CONCLUSIONS: The observed zona binding inhibition might be ascribed to zona receptor blocking on the sperm surface or the inability of acrosome-reacted sperm to bind to the zona pellucida.

Acrosome↗

PubMed1996

Acrosomal morphology as a novel criterion for male fertility diagnosis: relation with acrosin activity, morphology (strict criteria), and fertilization in vitro.

OBJECTIVE: To determine the relationships between sperm acrosin activity, sperm morphology evaluated according to strict criteria, visually observed acrosomal morphology, and IVF rates. DESIGN: Prospective analytic study. Acrosin activity was determined on all semen samples together with a standard semen analysis. Emphasis was placed on sperm morphology and especially a novel criterion viz acrosome morphology (acrosome index) as recorded with bright field microscopy. SETTING: University-based tertiary care center. PATIENTS: Thirty-three couples undergoing IVF or GIFT with two or more metaphase II ova inseminated in vitro. MAIN OUTCOME MEASURE: In vitro fertilization rates of inseminated ova. RESULTS: Strong correlations were found between acrosome index, normal sperm morphology, and IVF rates. An acrosome index cutoff value could be established at > 10% normal acrosomes for IVF rates of > or = 50% (sensitivity and specificity = 100%) and an acrosin activity cutoff value at > 18 microIU/10(6) sperm. A multiple linear regression analysis showed that the acrosome index and acrosin activity added a significant contribution to the explanation of the variation in the fertilization rates. CONCLUSIONS: A strong positive correlation was found between acrosome index and IVF rates. Although the numbers of the study are small, the results indicate that the acrosome index possibly may be regarded as an additional tool in the prediction of IVF outcome and especially may be of value in the group of men with severe teratozoospermia, i.e., < or = 4% morphologically normal spermatozoa.

Acrosin↗

PubMed1996

A prospective study on the predictive value of normal sperm morphology as evaluated by computer (IVOS).

OBJECTIVE: To evaluate the IVOS (Hamilton Thorne Research Version 2.1 Dimension Program, Beverly, MA) system's ability to predict fertilization in vitro in a prospective study. DESIGN: Hospital-based academic ART program. PATIENTS: Eighty patients from the IVF-GIFT program were evaluated. The same semen sample was analyzed on a day-to-day basis by both laboratory (manual method) and the computerized system for percentage normal morphology, concentration/mL, motility, and forward progression. Only patients with two or more metaphase II (MII) oocytes available were allowed into the study and excluded where the male partner had antisperm antibodies or qualified for intracytoplasmic sperm injection (<500,000 motile spermatozoa obtained after glass wool separation). STATISTICAL ANALYSIS: Logistic regression analysis was used to study predictors of fertilization in vitro. RESULTS: Three hundred thirty-eight oocytes were obtained from 80 patients of which 239 fertilized. The logistic regression analysis of the manual method (percentage normal morphology) and IVOS indicated that both were predictors of fertilization. Sperm morphology as evaluated by IVOS in patients with <10 x 10(6) motile spermatozoa/mL retrieved after swim-up was a significant predictor of fertilization as was the number of oocytes obtained. Thus, the more oocytes obtained in the lower morphological groups, the better the chance of fertilization. The fertilization rate in the morphology group of 0% to 4% normal forms was 45.6% (37/81) and in the group >14% normal forms was 85.2% (69/81). CONCLUSIONS: It was shown that in patients where </= 10 x 10(6) motile spermatozoa were obtained, the role of morphology (evaluated by IVOS) as well as the number of oocytes were important predictors of fertilization. The computer can assist to identify these patients with a poor prognosis for fertilization.

Computer Simulation↗

PubMed1996

Inhibition of G protein in human sperm and its influence on acrosome reaction and zona pellucida binding.

OBJECTIVE: To evaluate human sperm acrosomal status, zona pellucida (ZP)-binding capacity, and sperm motion characteristics after treatment with pertussis toxin followed by exposure to increasing concentrations of solubilized human ZP. DESIGN: Prospective analytical study. SETTING: Normal human sperm donors in an academic research environment. INTERVENTION: Sperm were prepared with a wash and swim-up method and treated with a final concentration of 100 ng/mL pertussis toxin. Acrosomal status were determined using a Pisum sativum agglutinin-fluorescien-isothiocyanate method after exposure of sperm to 0.25, 0.5, 0.75, and 1.00 ZP/microL solutions of human ZP. Zona binding potential was recorded using intact zona-binding assays. Motion characteristics were recorded with a semen analyzer. MAIN OUTCOME MEASURE: Percentage acrosome-reacted sperm, number of zona-bound sperm, and sperm motion parameters. RESULTS: Spermatozoa treated with 100 ng/mL pertussis toxin, followed by ZP-mediated acrosome reaction induction, showed a significant decrease in the percentage of acrosome-reacted sperm compared with untreated controls. Motion characteristics of 3-hour capacitated sperm after treatment with either phosphate-buffered saline (PBS) or pertussis toxin were not different. Pertussis toxin-treated sperm populations bound significantly more sperm to the ZP after 4 hours incubation compared with the PBS-control groups: 137.1 +/- 8.0 compared with 96.3 +/- 7.0 (mean +/- SEM). CONCLUSIONS: The data support the concept of the controlling mechanism and importance of G proteins during the ZP-mediated acrosome reaction. Intact acrosome correlate with and are needed to ensure tight zona binding.

Acrosome↗

PubMed1995

Influence of elevated pH levels on structural and functional characteristics of the human zona pellucida: functional morphological aspects.

PURPOSE: A total of 86 fresh and salt-stored immature human oocytes derived from postmortem ovarian tissue were used for this study. METHODS: Oocytes were randomly incubated either in synthetic human tubal fluid medium (untreated zonae) or in a chemically defined medium (treated zonae). RESULTS: Sperm binding experiments using hemizona assay conditions exhibited a 10-fold increased binding of sperm to treated compared to untreated oocytes (272.7 +/- 43 versus 24.3 +/- 15 sperm bound, respectively; P < 0.0001). pH recordings during incubation showed elevated pH levels of 8.1 compared to pH 7.2 among treated and untreated zonae, respectively. Ultrastructural examination showed a spongy appearance of the surface of treated zonae, whereas untreated zonae appeared compact with smooth surface. CONCLUSIONS: The marked increase in sperm binding among treated zonae, together with the ultrastructural findings, suggest that the altered zona surface enhances sperm binding. The physiological maturational process of the zona pellucida might be manipulated in vitro, thus increasing sperm binding to the zona.

Culture Media↗

PubMed1995

Sperm morphology: assessing the agreement between the manual method (strict criteria) and the sperm morphology analyzer IVOS.

OBJECTIVES: To correlate the percentage normal morphology reported by different observers and a computerized method (IVOS; Hamilton-Thorne Research, Beverly, MA) on a slide-by-slide basis using strict criteria: [1] Experienced observer (T.F.K.) versus experienced observer (R.M.), [2] experienced observer (T.F.K.) versus sperm morphology analyzer (IVOS), and [3] repeatability of normal and abnormal cells (IVOS). DESIGN SETTING, PATIENTS: Slides from 30 different patients from the Tygerberg IVF program were selected randomly. Microscopic fields and sperm cells were chosen randomly and percent normal morphology was recorded (objectives 1 and 2). The same slides were used and a cell-by-cell repeatability was done as outlined (objective 3). RESULTS: Experiment 1 (objective 1): there was no significant bias between T.F.K. and R.M. The limits of agreement were 8.6% and -7.3%. The SDs were not significantly different (P = 0.1283). The Spearman correlation coefficient between readers was 0.83. Experiment 2 (objective 2): the same findings were reported but the limits of agreement were 12.1% and -15.5%. The Spearman correlation coefficient was 0.85. The limits of agreement was tighter below 20% normal forms (+8.4 and -6.6). Experiment 3 (objective 3) (repeatability): 255 cells were analyzed three times in succession. Estimating pairwise agreement, the kappa statistic for the pairs are 0.85, 0.80, and 0.85, respectively, which compares favorably with the second canonical moment of 0.8329 (kappa = 0.83). DISCUSSION: The computer's ability to classify normal morphology per slide is promising. Below 20% normal forms, the limit of agreement is tight. Because of the 6% higher reading compared with the manual method, different thresholds possibly will be developed to identify subfertile from fertile patients. The computer gives excellent repeatability of normal and abnormal cells. Based on results obtained, this system can be of clinical value both in IVF units and andrology laboratories but more clinical data is required in this field.

Cytological Techniques↗

PubMed1995

Fluorescent in situ hybridization evaluation of human Y-bearing spermatozoa separated by albumin density gradients.

OBJECTIVE: Fluorescent in situ hybridization of decondensed sperm nuclei was employed to evaluate the effectiveness of the Ericsson albumin gradient enrichment technique of Y-bearing sperm. DESIGN: Mature human spermatozoa were separated from donor- and randomly selected patient semen using three different albumin density layers. Sperm nuclei, in the albumin layer with highest density, were decondensed and the percentage Y-bearing sperm was determined by fluorescent in situ hybridization. RESULTS: A slight, although statistically significant, increase in percentage Y-bearing sperm was observed. CONCLUSION: Although statistically significant, the clinical relevance of the small increase in Y-bearing sperm remains uncertain. A randomized controlled clinical study should help to clarify the above laboratory results.

Cell Nucleus↗

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↗

PubMed1994

Fertilization efficiency of morphologically abnormal spermatozoa in assisted reproduction is further impaired by antisperm antibodies on the male partner's sperm.

OBJECTIVE: To investigate the influence of antisperm antibodies on the sperm surface on the outcome of IVF and GIFT. DESIGN: Matched controlled retrospective review of two large series. SETTING: Reproductive endocrine divisions of two level-three academic centers. PATIENTS: Twenty-nine male factor patients (38 IVF cycles) showing positive antisperm antibodies on the sperm by immunobead test treated by IVF at the Norfolk program and 56 similar patients (57 cycles) treated by GIFT at the Tygerberg program. Twenty-nine male factor patients (29 IVF cycles) with negative antisperm antibodies screening matched by wife's stimulation protocol and baseline semen analysis characteristics were used as controls in Norfolk; 56 GIFT patients (56 GIFT cycles) matched similarly were the Tygerberg controls. Study and control groups were divided according to sperm morphology pattern in normal, good, and poor prognosis subgroups for comparison. MAIN OUTCOME MEASURES: Fertilization rate of preovulatory oocytes used in IVF in Norfolk and of GIFT supernumerary preovulatory oocytes in Tygerberg; total and term pregnancy rates (PRs) and abortion rates. RESULTS: Fertilization rate was significantly lower in the IVF (41.9% +/- 2.8%) as well as in the GIFT (26.8% +/- 3.8%) (mean +/- SE) study groups than in the respective control groups (73.1% +/- 3.9% and 61.8% +/- 3.9%). Total and term PRs in IVF per cycle (21.1% +/- 6.6%; 13.2% +/- 5.5%) and per transfer (23.5% +/- 7.4%; 14.7% +/- 6.1%), and in GIFT (25.0% +/- 5.8%; 19.6% +/- 5.3%) in the study groups were also lower when compared with their control counterparts (IVF per cycle: 62.1% +/- 6.2% and 41.4% +/- 6.0%; IVF per transfer: 41.9% +/- 2.0% and 27.9% +/- 1.9%; GIFT: 31.6% +/- 6.2% and 28.1% +/- 6.0%), but the difference did not reach statistical significance. Abortion rates were similar in the IVF study group (37.5% +/- 17.1%) and its control groups (39.9% +/- 11.5%). The abortion rate in the GIFT study group was 14.3% +/- 9.4%, and no abortions were recorded in the control group (not significant). CONCLUSIONS: The presence of antisperm antibodies on the sperm surface per se impairs the outcome of assisted reproduction, mainly in terms of fertilization rate of preovulatory oocytes, and possibly in terms of total and term PRs. This holds true regardless of the impact of other semen parameters, particularly the morphology of the sperm within the semen sample.

Abortion, Spontaneous↗

PubMed1994

Development of a predictive model for optimal zona pellucida binding using insemination volume and sperm concentration.

OBJECTIVE: To develop a predictive model under hemizona assay (HZA) conditions for human spermatozoa concentrations and insemination volume for optimum zona pellucida (ZP) binding. DESIGN: Analysis of 20 different insemination volumes for zona binding and sperm morphology under HZA conditions. SETTING: Reproductive biology unit, tertiary medical center. PATIENTS: Four proven fertile sperm donors. MAIN OUTCOME MEASURES: 5-, 20-, 50-, 80-, and 100-microL droplets were analyzed with four different concentrations of 0.5 x 10(6), 1.0 x 10(6), 2.0 x 10(6), and 4.0 x 10(6) cells/mL to determine the number of sperm bound to each hemizona. Fifteen hemizonae were used for each insemination volume or microdroplet. Response surface regression model with volume and concentration as the regressor variables has been used. RESULTS: The response surface of binding for the factors concentration and volume showed nonlinear association. A formula, indicating the optimal sperm insemination volume for maximum sperm binding to the ZP, Vmax = -(b1 + b5c)/2b6c, is described. The transformed data indicated 60 microL containing 4 x 10(6) sperm/mL to be optimal. Although morphology of zona spermatozoa is superior compared with seminal and postswim-up samples, no difference among the percentage of the normal morphology in different microdroplets could be demonstrated. CONCLUSION: Optimal volume for the obtained concentration of spermatozoa from a patient can be calculated and therapeutically used for cases of severe oligozoospermic patients by microvolume inseminations in IVF practice.

Female↗

PubMed1994

The effect of pentoxifylline on sperm movement characteristics and zona pellucida binding potential of teratozoospermic men.

This study investigates the effect of pentoxifylline on sperm movement and zona pellucida binding. Spermatozoa from nine teratozoospermic patients (10.2 +/- 6% normal forms) were included in the study. Test samples were diluted with a 4 nM solution of pentoxifylline, and control samples with Ham's F-10 culture medium only. Zona pellucida binding potential was measured under hemizona assay conditions (HZA). Sperm motility was evaluated at the start (0 h post-swim-up) and end (4 h) of the HZA, under test-tube conditions and under HZA conditions (50 microliters droplet under oil). Motility parameters tested included the curvilinear velocity (VCL), straight line velocity (VSL) and linearity (LIN). Compared to the controls, pentoxifylline-treated samples showed an immediate stimulation of sperm motility, under test-tube conditions, with a significant elevation of VCL at 0 h incubation (102.77 +/- 14.4 versus control value 84.60 +/- 10.6 microns/s; P = 0.005), which however was reversed after 4 h incubation (73.16 +/- 4.6 versus control value 85.47 +/- 12.8 microns/s; P < 0.005). A decline in sperm motility from 0 to 4 h incubation was noted for all the pentoxifylline-treated samples, under both test tube conditions (VCL: 102.77 +/- 14.4 versus 73.16 +/- 4.6 microns/s, P < 0.005; VSL: 27.2 +/- 10 versus 10.66 +/- 2.2 microns/s, P < 0.005; LIN: 23.65 +/- 7.1 versus 11.86 +/- 1.8%, P < 0.005) and HZA conditions (VCL: 100.04 +/- 13.1 versus 76.00 +/- 7 microns/s, P < 0.005; VSL: 26.40 +/- 8.7 versus 9.14 +/- 4.5 microns/s, P < 0.005; LIN: 26.2 +/- 12 versus 11.05 +/- 4.3%, P < 0.005).(ABSTRACT TRUNCATED AT 250 WORDS)

Female↗

PubMed1993

A new computerized method of reading sperm morphology (strict criteria) is as efficient as technician reading.

OBJECTIVES: To compare the ability of a computerized method of sperm morphology with the manually recorded method in predicting in vitro fertilization (IVF) results, to compare results obtained by both methods, and to determine the intraobservation variability. DESIGN, SETTING, PATIENTS: Forty-three stained semen slide preparations from two large level-three academic institutions' reproductive endocrinology units (IVF programs) were blindly evaluated, and the sperm were classified into normal and amorphous forms. RESULTS: Experiment 1: Twenty-one slide preparations from the Tygerberg gamete intrafallopian transfer program were manually evaluated; the fertilization rates for the groups with < 14% and > 14% normal sperm forms were 33.3% (15/45 oocytes) and 76.6% (46/60 oocytes), respectively. Corresponding fertilization rates with FERTECH were 46.8% (30/64) and 75.6% (31/41). Experiment 2: Twenty-two slide preparations from the Norfolk IVF program were evaluated. The manual method reported a fertilization rate in the group with < 14% normal forms of 27.4% (14/51 oocytes) compared with 90.0% (127/141 oocytes) in the group with > 14% normal forms. Corresponding figures for the FERTECH method were 33.9% (18/53) and 88.4% (123/139), respectively. Experiment 3: When the 43 slide preparations were blindly evaluated using both methods, 84% of the FERTECH evaluations correlated well with the manual method and FERTECH ability to diagnose the subfertile male (< 14% normal forms) was 95% (sensitivity). Experiment 4: A total of 16 different slides (8 per group) were randomly selected and analyzed five times (100 cells per reading) by the computerized method. The slides were obtained from men with normal sperm morphology of < 14% and > 14% as classified by the manual method. In the first group (< 14%) 97.5% (39/40) of the readings classified the sperm in the proper category, whereas in the second group (> 14%) 95% (38/40) of the cases were correctly identified. CONCLUSION: Using strict criteria for morphology evaluation, there is a positive and significant correlation between FERTECH evaluation and manual assessment. The reproducibility of the computerized method and the ability to distinguish between fertile and subfertile groups using those criteria are good.

Evaluation Studies as Topic↗

PubMed1993

The hemizona assay: its role in identifying male factor infertility in assisted reproduction.

OBJECTIVES: To identify male factor infertility among a group of patients in an assisted reproductive program (phase 1) and to evaluate the hemizona assay (HZA) in the diagnosis and prognosis of such a program (phase 2). DESIGN: The IVF performance of normal gametes in the Tygerberg program were critically evaluated. Female patients were classified as pure tubal factor infertility, having a normal FSH:LH ratio on day 3 of the menstrual cycle. All participating women produced three or more preovulatory oocytes at retrieval and were inseminated with sperm considered normal by all present diagnostic criteria. The total and normal fertilization rate thresholds were defined in that group. Using those thresholds, couples tested for sperm binding in the HZA (n = 48) were used and divided into two groups according to their fertilization rates, namely group 1, low fertilization (< 55%) and group 2, normal fertilization (> 55%). SETTING: University-based tertiary care center. PATIENTS: Ninety-nine couples (589 oocytes) with pure tubal factor infertility and normal male factor were used in phase 1. Forty-eight couples with normal and abnormal male factors that had both HZA performed and IVF treatment were included in phase 2. RESULTS: Investigation of the performance of normal gametes in 99 couples (589 oocytes) revealed the total fertilization rate (total number of oocytes fertilized/total number of oocytes inseminated) was (mean +/- SD) 88.6% +/- 16.8% and the normal fertilization rate (total number of oocytes with normal fertilization/total number of oocytes inseminated) was 81.3% +/- 22%. The minimum total fertilization rate that can be considered normal in the Tygerberg program using mean--2 SD is therefore 55% and for normal fertilization rate is 37%. The group with low fertilization rate (< 55%) showed a mean hemizona index (HZI) significantly lower; nevertheless, the distribution overlapping indicates a low discriminating power of the HZA. A sensitivity of 75% and a specificity of 75% were found; the positive and negative predictive values were 81% and 68%, respectively. CONCLUSIONS: The results indicated the HZA and HZI contribute important information and can serve in conjunction with other semen characteristics as useful tools during the diagnosis of the male factor in assisted reproduction.

Chorionic Gonadotropin↗