Sperm functional changes and fertilization in vitro in co-culture with human skin fibroblasts

This study was undertaken to evaluate the effects of human skinfibroblast monolayers on human sperm function and fertilizationin vitro. Sperm function was evaluated using the hamster oocytepenetration assay (HOPA) and zona binding assay (ZBA) in mediumalone and in co-culture with human skin fibroblast monolayersand suspensions. The ZBA was also studied in fibroblast conditionedmedium and in bovine oviduct cell monolayers and suspensions.Fertilization was measured both in in-vitro fertilization (IVF)couples with a normal semen analysis (first study; randomized)and in IVF couples with subnormal semen analysis (second study;each patient served as its own control). The HOPA results werenot significantly different with or without fibroblasts. Inall co-culture situations and in conditioned medium the ZBAscored significantly lower than medium alone. No significantdifferences with respect to IVF were observed between the co-cultureand the control group in either study. The mean fertilizationrate per patient was 60% in the group with normal semen analysisand 25% in the group with abnormal semen analysis. From thisstudy we conclude that although co-culture with human skin fibroblastsand epithelial cells influences the results of some sperm functiontests, it does not influence fertilization in vitro.     

Infertility: The prognosis for assisted conception treatment after unexpected failure of fertilization in vitro: a comparative study

The relative prognosis for further assisted conception treatment(without micro-injection) after initial unexpected failure offertilization in apparently favourable couples undergoing in-vitrofertilization (IVF) treatment was assessed. After their firstcycle of treatment, 481 consecutive couples were grouped accordingto their fertilization (including cleavage) rate per oocyteinto five bands. Proportions of couples proceeding to furthercycles of treatment by IVF or gamete intra-Fallopian transfer(GIFT) and resulting fertilization and pregnancy rates werecompared. Pregnancy rates in the first cycle of treatment weresignificantly related to fertilization rate. The fertilizationrate was zero in 13 couples (3%) and only 1–24% in 18(4%). There were no significant differences between these groupsin the proportions proceeding to further treatment (31, 50%)compared with others (overall 37%, including some treated byGIFT), or in their median fertilization rates (75, 60% comparedwith 67% – IVF cycles only), pregnancy rates (20, 38%of cycles compared with 37% – IVF or GIFT) or birth rates(20, 38% of cycles compared with 31% – IVF or GIFT). Amongstcouples whose initial fertilization rate was 50% there wasno fertilization in 4% of subsequent IVF cycles. We concludethat in couples with well defined favourable conditions, includingtests of sperm function for assisted conception treatment, whohave unexpected failure of fertilization, the prognosis forfurther treatment remains favourable without resort to morecomplex investigations or micro-injection methods. Such failureoccurs infrequently and generally as a random event, and shouldhave no appreciable effect on life-table calculation of cumulativepregnancy and birth rates in this group of patients.     

Andrology: Incubation of spermatozoa from asthenozoospermic semen samples with pentoxifylline and 2-deoxyadenosine: variability in hyperactivation and acrosome reaction rates

We examined hyperactivation and acrosomal loss in asthenozoospermicpatients with a history of failed in-vitro fertilization (IVF).After selection by a Percoll gradient, spermatozoa were incubatedwith 3.6 mM pentoxifylline (PTX), 3.0 mM 2-deoxyadenosine (2-DXA)or both. Hyperactivation and ionophore A-23187-induced acrosomereaction were assessed immediately after sperm treatment andagain after 180 min. In all groups studied, the mean hyperactivationrates were found to be low. No significant differences werenoted between assessments immediately after treatment and 180min later, except after treatment with both PTX and 2-DXA. Themean hyperactivation rates were found not to improve as a resultof either PTX or 2-DXA, while the combination of both PTX and2-DXA revealed a significant enhancement of total hyperactivation.When individual hyperactivation rates between control and treatedsperm samples were compared, large differences in response wereobserved. Some sperm samples showed a marked increase in hyperactivationwith one treatment, while another treatment led to a decrease.Acrosome reaction rates assessed immediately after ionophoreA-23187 stimulation were found not to be significantly differentfrom those assessed 180 min later. No significant effect couldbe demonstrated for either treatment, although, here too, markedinterindividual variations were noted. It was concluded thatan unselective use of PTX, 2-DXA or both compounds together,may restore sperm function in certain of these patients, andperhaps improve fertilization in vitro, but in others it mayproduce no change or may even be detrimental to sperm function.     

A self-migration method for preparation of sperm for in-vitro fertilization

Human sperm samples (n = 211) were prepared for in-vitro fertilization(IVF) and embryo transfer by a self-migration procedure in Earle'smedium containing highly purified hyaluronic acid (Hya) (MW3 000 000) included to increase the viscosity of the medium.The method resulted in the recovery of a significantly higherpercentage of motile spermatozoa compared with the traditionalcentrifugation method, 87.5 ± 0.9% versus 76.1 ±1.3% (P < 0.001). When comparing media with and without Hyain the selfmigration method for preparation of normal spermsamples, the media containing Hya resulted in the recovery ofa significantly higher percentage of motile spermatozoa, 89.0± 0.8% versus 73.8 ± 2.0% (P < 0.001). In agroup of 80 consecutive couples entering our IVF programme,sperm samples from 44 of the men were allocated at random forthe self migration method in medium containing Hya and spermsamples from 36 men for preparation by centrifugation and swim-up.Significantly more pregnancies were achieved in the group preparedin medium containing Hya. It is concluded that self-migrationof sperm in a medium containing Hya is simple and rapid, andresults in a high recovery of motile spermatozoa which can beused for in-vitro insemination of human oocytes with favourableresults.     

Andrology: Pentoxifylline-enhanced acrosome reaction correlates with fertilization in vitro

To evaluate the possible effect of pentoxifylline on the acrosomereaction (AR) and its correlation with in-vitro fertilization(IVF), sperm samples obtained from 51 patients who underwentIVF treatment were studied. Acrosome reactions were evaluatedas spontaneous, pentoxifylline-treated and calcium ionophore(A23187) induced, before and after treatment. The correlationof AR with fertilization in vitro in spermatozoa pre-treatedwith pentoxifylline was sought. In cases with failure or verylow fertilization rate (10%) in their previous trials, spermatozoaafter swim-up were treated before insemination. Spontaneousacrosome loss remained low even after treatment (mean ±SD: 8.18 ± 1.74%). Response to A23187 was enhanced significantly(P < 0.001) by pre-treatment with pentoxifylline in 33 controlcases (group A) in which fertilization in vitro was previouslysuccessful without this treatment. Patients with at least twoepisodes of failed fertilization were divided into two groups.In 11 cases (group B), the IVF rate was improved significantly(P < 0.001) by the treatment. This was not observed in sevencases (group C) in which the treatment induced no increase inIVF rate. We achieved nine (27.3%) pregnancies in group A andfive (45.4%) pregnancies in group B. This study demonstratedthat pentoxifylline enhanced A23187 induced the acrosome reactionand this effect was correlated with improvement in IVF rate.     

Protein tyrosine phosphorylation, hyperactivation and progesterone-induced acrosome reaction are enhanced in IVF media: an effect that is not associated with an increase in protein kinase A activation

Sperm capacitation is a prerequisite for successful in vitro fertilization (IVF) and therefore a focus of sperm preparation in IVF laboratories. The technology of IVF is, therefore, potentially valuable in advancing our understanding of the molecular processes that occur during sperm capacitation. We have investigated sperm capacitation induced by a commercial IVF medium compared to that occurring in standard capacitating medium (CM) typically used in a nonclinical setting. Percoll-washed spermatozoa were resuspended in Cook Sydney IVF medium, Cook Sydney IVF sperm buffer, Earle's balanced salt medium (capacitating medium) or a modified Earle's balanced salt medium [non-capacitating medium (NCM)] for up to 120 min at 37 degrees C and, if applicable, in the presence of 5% CO2 in air. Sperm protein kinase A (PKA) activity, PKA-dependent serine/threonine phosphorylation, tyrosine phosphorylation, hyperactivation and progesterone-induced acrosome reaction were evaluated. IVF medium was shown to accelerate sperm capacitation (compared with capacitating medium) as determined by tyrosine phosphorylation, sperm hyperactivation and progesterone-induced acrosome reaction. This effect was not associated with enhanced activation of PKA or increased levels of serine/threonine phosphorylation. In contrast, IVF sperm buffer (used for sperm preparation) did not stimulate sperm capacitation when incubated for up to 90 min. We have shown that different capacitating media vary strikingly in their efficacy and that this difference reflects activation of a pathway other than the well-characterized activation of soluble adenylyl cyclase/cAMP/PKA.     

Andrology: High fertilization prediction by flow cytometric analysis of the CD46 antigen on the inner acrosomal membrane of spermatozoa

The study was set up to determine the relationship between thehuman sperm acrosome reaction and fertilization in couples undergoingroutine in-vitro fertilization (IVF) treatment. Prospectivedata analysis was carried out on all IVF patients during a 6month period. Exceptions were those patients having insufficientsperm concentration to allow both acrosome reaction determinationand insemination. The main outcome measures were the predictionof fertiliza tion in IVF patients using flow cytometric analysisof the spontaneous and ionophore-induced acrosome reaction [givingthe acrosomal response to ionophore challenge (ARIC) score]in the male partner's spermatozoa versus standard analyticalmethods of sperm motion parameters and morphology. Stepwiselogistic regression indicated only two independent factors predictiveof fertilization: ARIC score (x² = 109.6, P < 0.0001) andpost-Percoll % motility (x² 8.8, P < 0.003). Of patientswith an ARIC score of >10, 92% had >30% of oocytes fertilized;100% of patients with an ARIC score of <10 had <30% fertilizationof oocytes. The sensitivity and specificity of the assay systemwere 1.00 and 0.82 respectively. The results would indicatethat the ARIC test as measured by flow cytometric analysis ofCD46 binding is a sensitive and specific assay for use in theprediction of fertilization in IVF patients, thus enabling directchannelling of those patients with ARIC scores of <10 intothe more invasive micro-assisted fertilization schemes.     

Andrology: Cystic fibrosis mutations impair the fertilization rate of epididymal sperm from men with congenital absence of the vas deferens

One of the limiting factors for the successful treatment ofmale sterility due to congenital absence of the vas deferens(CAVD) is the low (<20%) and extremely unpredictable rateof in-vitro fertilization of their epididymal spermatozoa. Therecent demonstration that CAVD is a mild form of cystic fibrosis(CF) disease, almost exclusively involving the genital tract,has prompted us to investigate the hypothesis of whether therecould be an association between particular CF genotype mutationsand the in-vitro performance of epididymal sperm. In this study63 patients with surgically confirmed diagnosis of CAVD undergoingmicrosurgical epididymal sperm aspiration (MESA) and in-vitrofertilization (IVF) were evaluated. The genetic screening wascarried out on DNA extracted from peripheral lymphocytes andamplified by the polymerase chain reaction. A total of 12 mutationsin the cystic fibrosis transmembrane regulator gene (CFTR),representing 90% of the total known CF mutations, were tested.The presence or absence of mutations, as well as the type ofmutation found, was correlated with the IVF and pregnancy rates.Of the 63 patients examined, 40 (64%) were positive to the CFscreening and 23 were negative. In three cases no spermatozoawere found because of rete testis blockage. The difference infertilization rates between patients testing positive and negativewas highly significant (P = 0.000003), 13 versus 23%, respectively.In order to pinpoint which mutation could account for the mostdeleterious effect on the IVF results, we analysed these bykeeping the patients separated by CF mutations. Three majorgroups were identified: group I was formed by 21 patients (52%of the positive) with Delta F508, the most common and lethalof the CF mutations, as the only detectable. Group II was formedby 18 patients with various other CF mutations, including fourcompound heterozygotes (three with Delta F508/R117H and onewith R553X/R117H) and one homozygote (R117H/R117H). Group IIIwas formed by 21 patients in whom no mutations could be detected.The results showed a very low fertilization and pregnancy rateper patient (7 and 10%, respectively), in group I, while ingroups II and III the fertilization and pregnancy rates weresimilar (group II, 21 and 39%, respectively; group III, 23 and48%, respectively). The differences in the fertilization andpregnancy rates between groups I and III were statisticallysignificant (P < 0.000001 and P < 0.01, respectively),while there was no difference between groups II and III. Inconclusion, this study demonstrates a correlation between CFmutations and IVF capacity of epididymal spermatozoa retrievedfrom men with CAVD. Men with Delta F508 not opposed to knownCF mutations give the poorest IVF outcomes; also at repeatedattempts their pattern of poor fertilization is maintained,perhaps reflecting sperm intrinsic biochemical defects (alterationof chloride channels). These data open a new avenue in the interpretationof the IVF results from men with CAVD.     

Andrology: A new predictive test for in-vitro fertilization based on the induction of sperm acrosome reaction by N-acetylglucosamine-neoglycoprotein

Neoglycoproteins with N-acetylglucosamine residues (BSA-GIcNAc)induced specifically the acrosome reaction (AR) in human spermatozoa.Our objective was to investigate the relationship between thisphenomenon and the in-vitro fertilization (IVF) rate. Spermsuspensions from IVF protocols were incubated with BSA-GlcNAc(t), using calcium ionophore (i) or medium alone (c) as positiveor negative controls. When the normalized AR percentage ratio(STIM) (%ARt-%ARc): (%ARi-%ARc) was compared with fertilizationrate in 31 couples from our IVF programme, a positive correlationwas found (r = 0.46, P < 0.01). The fertilization rate inpatients with STIM 0.2 was higher than in non-responders (STIM< 0.2); 72 ± 7% compared with 5 ± 3%. The overallpredictive value of this test for adequate fertilization rate(>30%) was 87%, sensitivity 91% and specificity 78%. Falsepositives were 9% and false negatives 22%. For successful fertilizationrates (>60%), the results were: overall predictive value,84%; sensitivity 100%; specificity 64%. False positives were23% and no false negatives were found. The results indicatedthat the induction of AR in human spermatozoa by GlcNAc-neoglycoproteinscould be used to predict their fertilizing ability in vitro.     

Andrology: Intracytoplasmic sperm injection does not require special treatment pf the spermatozoa

In order to assess whether specific treatment of spermatozoais required prior to intracytoplasmic sperm injection (ICSI),three methods of sperm preparation were compared in this study.These three methods were (A) incubation of spermatozoa withpentoxifylline (PTX) and 2-deoxyadenosine (DOA), (B) electroporationfollowed by incubation in medium with PTX, and (C) no furthertreatment with the Percoll gradient. Controlled comparisonswere carried out between method A and method B in 21 patients,and between method A and method C in 32 patients. There wasno difference in the rates of fertilization and embryo cleavagewhen ICSI was done with spermatozoa treated by procedures A,B or C. Furthermore, the sperm selection procedure prior toICSI was done in two different media: T6 medium containing 1.78mM CaCl₂·2H₂O and a final washing step after the Percollgradient in T6 medium containing 5.0 mM CaCl₂·2H₂O, andEarle's medium containing 1.78 mM CaCl₂·2H₂O. The resultsof ICSI on sibling oocytes from 12 patients revealed no differencein the fertilization and embryo cleavage rates between the twodifferent media used during the sperm selection procedures.In conclusion, it appears that high fertilization and pregnancyrates can be obtained in couples with severe male-factor infertilityby ICSI and that no special treatment of the spermatozoa priorto ICSI is required.     

Treatment of severe male immunological infertility by intracytoplasmic sperm injection

A total of 29 infertile couples (group A) with male antispermantibodies detected by the mixed antiglobulin reaction (MAR)and partly by flow cytometry (n = 21) were treated using anintracytoplasmic sperm injection (ICSI) technique to assistfertilization. In all, 22 of them had shown a poor fertilizationrate (6%) in previous in-vitro fertilization (IVF) treatments.The fertilization and cleavage rates in ICSI, 79 and 89% respectively,were similar to those in a MAR-negative group (group B; n =20) injected because of male infertility (68 and 93% respectively).A third group (group C; n = 37) with male immune infertilitywas treated by conventional IVF. All these couples had at leastone oocyte fertilized, but the overall fertilization rate (44%)in group C was significantly poorer (P < 0.001) than thatin the two ICSI groups. However, the embryo quality was lowerin group A compared with that in the other groups. A total of13 pregnancies resulted in group A (46%), of which five endedin miscarriage. None of the six pregnancies (30%) in group Baborted during the first trimester. These results reveal, forthe first time, that ICSI offers a good chance of fertilizationfor couples with male immunological infertility. However, post-fertilizationevents may compromise these results because of factors not yetclearly understood.     

The hypo-osmotic swelling test and the sperm mucus penetration test in determining fertilization of the human oocyte

The usefulness of the hypo-osmotic swelling (HOS) test and thesperm mucus penetration (SMP) test as sperm function tests forin-vitro fertilization was analysed in 56 couples. Using logisticregression analysis only the SMP test was independently relatedto fertilization (P = 0.004), no false negative results wereobtained, i.e. no fertilization if sperm from the ejaculatefailed to penetrate mucus. The HOS test was of no predictivevalue. The results justify a further examination of the SMPtest in other IVF centres.     

The use of Albuminar 5 (TM) as a medium supplement in clinical FVF

Human serum and Albuminar 5 (A5) were compared as medium supplementsto Earle's solution containing pyruvate in clinical IVF. One-hundredpatients in each group showed a fertilization rate of 60% withserum and of 62% with A5. The overall pregnancy rates in theserum and A5 groups were 20 and 24%, respectively. The incidenceof failed fertilization (6–7%) and of multipronucleateoocytes (4–5%) was similar in both groups. At 37°C,sperm survived less well in A5 although the rate of fertilizationwas not reduced. Blastocyst formation was not seen in ‘spare’embryos grown in vitro in medium containing 15% v/v A5.     

Fertilization in vitro of human oocytes by spermatozoa collected in different stressful situations

It has been shown that semen quality is impaired in couplesundergoing in-vitro fertilization (IVF), probably due to stress.A possible effect of stress on the ability of spermatozoa tofertilize human oocytes in vitro was analysed in the presentstudy composed of 26 couples with normozoospermic men undergoingIVF. A semen sample was obtained during the infertility work-upand was cryopreserved (sample 1). A second sample (sample 2)was provided after oocyte retrieval during the IVF cycle. Sample1 was thawed and both samples were washed and preincubated foroocyte insemination. One-hundred-and-five oocytes were inseminatedusing thawed sample 1, and 120 with sample 2.Semen parameterssuch as density, progressive motility and percentage of abnormalforms were compared between sample 1, before and after freezing,and sample 2. Only motility was significantly (P<0.01) decreasedby cryopreservation in sample 1, but no parameter was significantlydifferent when fresh sample 1 was compared to sample 2. Thefertilization rate was 78.6% using sample 1 in comparison to87.5% when sample 2 was employed (not significant, NS). Cleavagerates were 77.7 and 89.7%, respectively (NS). A group of fivepatients undergoing IVF who needed donor semen served as a controlfor the effect of sperm cryopreservation on IVF. In these cases,the donor was asked to provide a fresh sample. Half of thissample was frozen and thawed. Subsequently, fresh and thawedsamples were prepared for insemination and oocytes inseminatedeither with the fresh preparation (n=24) or the frozen and thawedspermatozoa (n=22). There was a significant (P<0.05) decreasein motility in the thawed sample, but fertilization and cleavagerates were not different. These data suggest that the stressfulsituation induced by IVF treatment in normozoospermic men doesnot affect the ability of spermatozoa to fertilize human oocytesin vitro. Cryopreservation of human spermatozoa before IVF maybe a good policy in couples especially suspected of being understress during this procedure.     

A follow-up study of sperm preparation for IVF by swim-up in a solution of hyaluronate

Spermatozoa prepared for in-vitro fertilization (IVF) by swim-up in a balanced salt solution containing hyaluronate gave rates of fertilization, cleavage and pregnancy which were not significantly different from those obtained with sperm prepared by swim-up in standard IVF medium followed by centrifugation. However, the content of prostaglandin F2alpha in the final sperm suspension was higher using hyaluronate but this seemed to be of no consequence for IVF. Thus, preparation of normal sperm samples for IVF may be simplified by performing swim-up in a balanced salt solution containing hyaluronate.     

Fertilization and early embryology: Absence of heat treatment of serum for culture medium supplementation does not adversely affect the outcome of in-vitro fertilization

This study was carried out to determine if not heat-treatingserum prior to use for medium supplementation adversely affectedin-vitro fertilization (IVF) of human oocytes. Morphologicallymature human oocytes derived from 135 patients undergoing IVFtreatment were studied. A total of 504 oocytes were incubated,inseminated and the resulting pronuclear oocytes cultured furtherin Earle's balanced salt solution (EBSS) supplemented with 10%non-heat-treated serum. Comparisons of fertilization rate andembryonic development were made between these and 687 controloocytes derived from the same patients but incubated, inseminatedand resulting pronuclear oocytes cultured further in EBSS supplementedwith 10% heat-treated serum. The fertilization rate of 74.4%(375/504) of oocytes handled in serum-supplemented medium thathad not been heat-treated was significantly better than therate of 67.7% (465/687) for controls (P< 0.0125). The proportionof pronucleate oocytes that cleaved was also significantly betterin the non-heat-treated serum group: 270/300 (90%) versus 307/375(81.8%) (P < 0.0025). There was no significant differencein the proportion of embryos with four or more cells at thetime of embryo transfer. The results show that the absence ofheat treatment of serum used to supplement culture medium hasno adverse effect on the fertilization rate and short-term embryodevelopment in vitro; hence we suggest that serum heat treatmentis an unnecessary procedure and could be abandoned.     

Sperm DNA integrity assessment in prediction of assisted reproduction technology outcome

BACKGROUND: The sperm chromatin structure assay (SCSA) has beensuggested as a predictor of fertility in vivo as well as invitro. The available data however, have been based on limitednumbers of treatments. We aimed to define the clinical roleof SCSA in assisted reproduction. METHODS: A total of 998 cycles[387 intrauterine insemination (IUI), 388 IVF and 223 ICSI]from 637 couples were included. SCSA results were expressedas DNA fragmentation index (DFI) and high DNA stainable (HDS)cell fractions. Outcome parameters were biochemical pregnancy(BP), clinical pregnancy (CP) and delivery (D). RESULTS: ForIUI, the odds ratios (ORs) for BP, CP and D were significantlylower for couples with DFI >30% as compared with those withDFI 30%. No statistical difference between the outcomes of ICSIversus IVF in the group with DFI 30% was seen. In the DFI >30%group, the results of ICSI were significantly better than thoseof IVF. CONCLUSIONS: DFI can be used as an independent predictorof fertility in couples undergoing IUI. As a result, we proposethat all infertile men should be tested with SCSA as a supplementto the standard semen analysis. When DFI exceeds 30%, ICSI shouldbe the method of choice.     

Clinical importance of zona pellucida-induced acrosome reaction and its predictive value for IVF

The study aimed to establish zona pellucida induced acrosome reaction response (ZIAR) among 35 couples with normal and G-pattern sperm morphology and repeated poor fertilization results during assisted reproduction treatment. ZIAR tests were performed using 0.25 zona pellucida/microliter co-incubated with spermatozoa for 60 min. Acrosome reactions were measured with FITC-PSA staining, and expressed as the difference between stimulated and unstimulated (spontaneous) sperm populations. Results were compared with IVF rates of metaphase II oocytes. Interactive dot diagrams divided the patients into two groups, i.e. ZIAR <15% and ZIAR > 15%, with mean fertilization rates of 49 and 79% respectively. The sensitivity and specificity for ZIAR results versus fertilization were 93 and 100% respectively. The area under the curve was 99% and the 95% confidence interval did not include 0.5 which implies that the ZIAR test is able to predict fertilization failure among IVF patients. In conclusion, the ZIAR test has diagnostic potential since it can assist the clinician to identify couples that will benefit from intracytoplasmic sperm injection therapy.     

Andrology: The ability of the hemizona assay to predict human fertilization in different and consecutive in-vitro fertilization cycles

The objective of this prospective study was to examine the abilityof the hemizona assay (HZA) to predict fertilization outcomeof mature, pre-ovulatory oocytes under in-vitro fertilization(IVF) conditions. Since a large number of patients were evaluatedover a long period, the power of the HZA to prognosticate fertilizationresults in the same and subsequent (consecutive) IVF cyclesof those same patients was assessed. For IVF, only metaphaseII oocytes were used. For the HZA, both fresh oocytes donatedby patients at the time of IVF and oocytes recovered from surgicallyremoved ovarian tissue (and salt-stored) were used, and bisectedby micromanipulation techniques. Matching hemizonae were co-incubatedeither with spermatozoa from the patient (test) or from a fertileman (control) for 4 h. The number of spermatozoa tightly boundto the zona was counted. Patients (n = 112) were divided intotwo groups based on HZA results (expressed as HZA index or HZI):HZI 30% (n = 72) and <30% (n = 40). The patients with HZI<30% had significantly lower fertilization rates in boththe HZA—IVF cycle and in subsequent cycles compared topatients with HZI 30% (P < 0.03). Linear discriminant analysisindicated the HZA to have a sensitivity of 84%, and positiveand negative predictive values of 85 and 70% respectively, forprediction of fertilization outcome in a total of 233 cycles.It was concluded that the HZA is a good predictor of fertilizationrate in vitro, and can be used in the IVF setting to supplyadditional clinical information in malefactor patients.     

Does treatment with testosterone undecanoate improve the in-vitro fertilizing capacity of spermatozoa inpatients with idiopathic testicular failure?(results of a double blind study)

Seventy-seven couples in whom conventional in-vitro fertilization(IVF)had remained unsuccessful because of low fertilization rateand abnormal sperm characteristics were given either testosteroneundecanoate 120 mg/day, or placebo during 3 months, after whicha new IVF treatment was applied under identical technical conditions.There were no significant changes in sperm characteristics amongthe treated and placebo couples and the fertilization rate showeda similar increase after treatment in both groups. No significantdifference in pregnancies occurred, with 32% pregnancies inthe placebo controls and 17%among couples treated with testosteroneundecanoate. It is concluded that testosterone undecanoate intakedoes not improve sperm characteristics, or the in-vitro fertilizingpotential, or pregnancy rate over those observed in the placebocontrols in cases with primary idiopathic testicular failure.