Infertility: Characterization of Tru-Trax in-vitro penetration testing of cervical mucus

To determine the distribution of results of an in-vitro cervicalmucus penetration system employing both human and bovine mucus(Tru-Trax) in a general infertility population, 133 couplesprospectively underwent in-vitro and post-coital testing ofcervical mucus. The distribution of Tru-Trax results in coupleswith normal semen analyses and Insler scores showed significantlygreater penetration in bovine (22.3 ± 6.0 mm) than inhuman mucus (20.3 ± 5.4 mm) (P < 0.001). However,the lower limit of the 95% confidence interval of the normalpopulation with either type of mucus was 10 mm. This limit wassignificantly lower than that described by the kit manufacturer.The predictive value for post-coital tests using human mucusin the Tru-Trax system was good (>90%) in all groups. Theoverall penetration into either human or bovine mucus was significantlycorrelated with the percentage of motile spermatozoa in thesemen sample (P < 0.001). In conclusion, in-vitro cervicalmucus penetration tests with the Tru-Trax system are relatedto in-vivo post-coital tests, although the lower limit of therange of anticipated results in the normal population was significantlylower than that described by the manufacturer. The ultimatevalue of this type of testing will await clinical trials whichevaluate clinical end-points such as pregnancy rates.     

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.     

Fertilization and early embrology: A sequential analysis of the effect of progesterone on specific sperm functions crucial to fertilization in vitro in infertile patients

The objective of these studies was to evaluate the modulatoryeffect(s) of progesterone on sperm functions crucial to fertilizationin infertile men with abnormal sperm parameters. A prospective,controlled study applying a sequential diagnostic analysis capableof identifying specific dysfunctions of the male gamete wasperformed. Patients (n = 14) were allocated to the study groupif they had a history of infertility of >1 year durationand after semen evaluation showed teratozoospermia (< 14%normal sperm forms as diagnosed by strict criteria) or terato-asthenozoospermia(< 50% progressive motility). After swim-up separation ofthe motile sperm fraction, the following functions were assessedwith and without previous exposure to progesterone (1.0 µg/ml):acrosome reaction (using Pisum sativum agglutinin), hyperactivatedmotility (using a computerized semen analyser), sperm-zona pellucidabinding (in the hemizona assay), sperm-zona pellucida penetration(in a sperm-zona penetration assay), and sperm-oocyte penetration(using the hamster zona-free oocyte/sperm penetration assay).Progesterone did not affect the percentage of acrosome-reactedspermatozoa after 1 or 3 h of incubation. Hyperactivated motilitywas significantly enhanced by progesterone after 1 h (12 ±4 versus 6 ± 2% in controls; P < 0.02). Although progesteronedid not affect sperm-zona binding, it significantly enhancedboth sperm-zona pellucida penetration (27 versus 12% in controls;P = 0.03) and sperm-oocyte penetration (15 versus 8% in controls;P < 0.05). Because those sperm functions enhanced by progesteroneare crucial to fertilization, the steroid may have value inthe treatment of some male-factor patients undergoing assistedreproductive therapy.     

In-vitro cervical mucus -- sperm penetration tests and outcome of infertility treatments in couples with repeatedly negative post-coital tests

The results of in-vitro cervical mucus—sperm penetrationtests and cross-hostility tests in 178 couples with repeatedlynegative post-coital tests were recorded. Using a protocol ofthree cycles with intra-uterine inseminations (IUI) followedby three cycles with ovulation induction + IUI, the associationbetween the cause of infertility, results of the in-vitro testsand the outcome of infertility treatment was investigated. Wefound that repeatedly negative post-coital tests are a goodindicator of a cervical mucus — sperm penetration problem.The crosshostility test clearly differentiates the abnormalfactor in this interaction, and a good performance of the donors'spermatozoa in the cervical mucus correlates with increasedpregnancy rate. In male factor infertility, failure of the husbands'spermatozoa to penetrate cervical mucus was not indicative ofa deficient fertilization potential in vivo. In these patientsa serious attempt should therefore be made to reverse the infertilityby treatment with IUI or ovulation induction + IUI before attemptingassisted reproduction. Women with polycystic ovaries and repeatedlynegative post-coital tests should be investigated for spermreceptivity of the cervical mucus. Low receptivity of the cervicalmucus may imply that endometrial receptivity and oocyte qualityare also low. Ovulation induction and not IUI alone should thereforebe used as the preferred mode of treatment to improve pregnancyrate.     

The diagnosis of male infertility--prospective time-specific study of conception rates related to seminal analysis and post-coital sperm--mucus penetration and survival in otherwise unexplained infertility

Infertile women without any inherent female infertility factorsand able to secrete normal cervical mucus were studied prospectivelyin relation to post-coital sperm—mucus penetration (PCT)and their partner's seminal analysis, excluding men with azoospermia.Time-specific cumulative conception rates calculated as forlife-table analysis were related to each measured seminal variableon routine analysis of 2–3 samples (volume, density, proportionwith progressive motility, and proportion with normal morphology);to various derivatives from combinations of these variables;to seminal findings after vital staining; and to the PCT results.The best seminal predictor of fertility was the motile normalsperm density (MNSD), the 18 month conception rates being 57.4%+ 4.6 (SE) and 30.2% + 5.9 (ratio 1.9, P < 0.001) above andbelow a derived threshold value of 4 x 10⁶/ml. The PCT led torates of 55.6% ± 4.3 and 14.9% ± 5.1 (ratio 3.73,P < 0.001) for positive and negative results, respectively.The PCT also gave rise to a significantly distinct intermediatepoor-psitive sub-group (conception rate 30.6% ± 9.0).Seminal analysis (the MNSD) did not affect the conception rateassociated with a positive PCT but helped to discriminate furtherwith a negative PCT (conception rates 22.5% ± 8.7 withan MNSD above 4 x 10⁶/ml versus 5.6% ± 4.8 below, P <0.05). The PCT was the single best predictor of fertility butseminal analysis (the MNSD) was of additional value after anegative PCT.     

The possible meaning of transferrin and its soluble receptors in seminal plasma as markers of the seminiferous epithelium

Transferrin (Tf) and soluble transferrin receptors (S-Tf-R)were measured by enzyme immunoassay in seminal plasma of 130semen samples. The mean concentration of S-Tf-R in cases withnormozoospermia was 10.4 IU/ml (95% confidence interval: 9.5–11.3)and it was significantly lower in patients with oligozoospermia(6.6, 95% CI: 5.8–7.5, P < 0.001), asthenozoospermia(8.5, 95% CI: 5.5–10.7, P < 0.05), azoospermia of primarytesticular origin (7.9, 95% CI: 6.1–9.6, P < 0.05)and post-vasectomy samples (5.9, 95% CI: 5.4–6.9, P<0.001). The concentration of S-Tf-R in post-vasectomy sampleswas lower than that in patients with azoospermia of primarytesticular origin (P< 0.05; positive likelihood ratio= 7at value of 8.3 IU/ml). S-Tf-R was positively correlated withmotile sperm concentration (r= 0.50, P< 0.0001), percentagemotility (r= 0.38, P< 0.001), percentage of normal forms(r = 0.43, P < 0.001), sperm linear velocity (r= 0.42, P<0.001), and ATP concentration (r= 0.67, P< 0.0001). Folliclestimulating hormone (FSH) was found to be negatively correlatedwith the concentrations of both Tf (r= -0.31, P< 0.05) andof S-Tf-R (r= -0.45, P< 0.01). The mean concentration ofTf in seminal plasma was 50.4 µg/ml (35.9–67.2)in samples with normozoospermia (n= 22), and the concentrationwas significantly lower in patients with oligozoospermia (P<0.05), azoospermia of testicular origin (P < 0.001), andpost-vasectomy samples (P< 0.001). Seminal Tf was correlatedwith motile sperm concentration (r = 0.36, P< 0.001), percentageof motile spermatozoa (r= 0.25, P < 0.05), linear velocity(r= 0.24, P< 0.05) and ATP concentration (r= 0.44, P<0.001). The concentration of Tf was positively correlated withthat of S-Tf-R both in cases with spermatozoa present (r= 0.66,P < 0.001), and in cases with azoospermia of testicular origin(r = 0.51, P < 0.05) but not in vasectomy cases. It is concludedthat S-Tf-R in seminal plasma is a marker of spermatogenesisand may give information on the presence or absence of spermatogeneticcells in cases with azoospermia. Further investigations areneeded to assess its usefulness for clinical practice.     

Fertilization and early embryology: Effects of embryo density and co-culture of unfertilized oocytes on embryonic development of in-vitro fertilized mouse embryos

We have evaluated the effects of embryo density and the co-cultureof unfertilized (degenerating) oocytes on the development ofin-vitro fertilized (IVF) mouse embryos. In experiment 1, groupsof one, five, 10 or 20 zygotes were cultured in 20 µldrops of modified human tubal fluid (HTF) medium for 168 h at38.7°C in 5% CO₂ and 95% air. As the embryo density increased,significantly (P < 0.05) higher rates of embryos reachedhatched blastocyst stage. In addition, the time required forhatching after IVF was significantly (P < 0.05) shortenedby the increase in embryo density. In experiment 2, 10 IVF zygoteswere cultured with or without 10 unfertilized (degenerating)oocytes in 20 µl drops of HTF medium. The rates of IVFembryos that developed to morula, blastocyst, expanded blastocystand hatched blastocyst stages were decreased significantly (P< 0.01) by culturing embryos with unfertilized oocytes comparedwith culturing embryos alone. In experiment 3, groups of oneor 10 IVF zygotes or 10 IVF zygotes plus 10 unfertilized oocyteswere cultured in 20 µl drops of HTF medium and the numberof cells per blastocyst was examined at 120 h after IVF. Increasingembryo density resulted in a significant (P < 0.05) increasein the number of cells per blastocyst. In contrast, the cellnumber of IVF embryos that developed to blastocyst decreasedsignificantly (P < 0.05) when they were cultured with unfertilizedoocytes. The results suggest that in-vitro development of IVFmouse embryos is enhanced by increasing embryo density and isimpaired by co-culture with unfertilized (degenerating) oocytes.     

Infertility: Dummy embryo transfer using methylene blue dye

The aim of this prospective work was to evaluate different cathetersand techniques used for embryo transfer. Studies were performedon 105 IVF patients before the start of treatment cycles. Eachpatient was used as her own control to study the expulsion ofmethylene blue (MB) dye after dummy embryo transfer. Group A(n = 35) underwent the test twice, before and after aspirationof the cervical mucus. Group B (n=30) underwent the test twicewith and without the presence of two air bubbles in the embryotransfer catheter. Group C (n = 40) underwent the test twiceusing two different catheters, the Wallace and the Craft catheters.The results showed that the dye was extruded at the externalos in 57% of the cases when the cervical mucus was not aspiratedcompared to 23% when the mucus was aspirated (P = 0.01). Thedye was extruded in 33% of the cases with air bubbles in thecatheter as compared to 27% when no air was present (P >0.05). When the Wallace catheter was used expulsion occuredin 25.5% compared to 77.5% when the Craft catheter was used.We concluded that using soft catheters and complete aspirationof cervical mucus significantly reduced the expulsion of thedye. The presence of air bubbles did not affect rate of expulsionof the dye.     

Relationships between motility parameters, morphology and acrosomal status of human spermatozoa

A total of 130 semen samples were examined for motility (bycomputer-assisted sperm analysis), morphology and acrosomalstatus. A high positive correlation was found between percentagesof normal forms and progressive motility in the whole semen(r = 0.539, P < 0.0001) as well as in the Percoll fraction(r = 0.702, P < 0.0001). Among the specific abnormalities,acrosome defects were most highly correlated with progressivemotility (r = –0.492, P < 0.0001, in the Percoll fraction).The percentage of total spontaneously acrosome-reacted spermatozoain the Percoll fraction was negatively correlated with the progressivemotility (r = –0.499, P < 0.0001) and with the percentageof normal forms (r = –0.430, P < 0.0001). Surprisingly,the percentage of total spontaneously acrosome-reacted spermatozoawas poorly linked with head abnormalities but displayed significantpositive correlations with the percentages of bent tails (r= 0359, P < 0.0001) and of coiled tails (r = 0371, P <0.0001). These data suggest that sperm defects are often linkedtogether, reflecting spermiogenesis and/or epididymal dysfunctions.     

Zona-opening of hamster oocytes: a comparative study using macro- and micro-manipulation methods

The present study compared a new macro-manipulation techniquefor zona-opening of hamster oocytes with existing micro-manipulationtechniques. In experiment I the zona pellucida of hamster oocyteswas partially opened with a 32 gauge steel needle (macro-manipulation)and these oocytes were then co-incubated with human spermatozoaat 36.5°C in 5% CO₂ in air for 3.5 h. Zona-free and zona-intactoocytes similarly inseminated served as controls. Of 113 oocytes,30 (26%) lysed following zona-opening with the macro technique.The sperm penetration rates of zona-opened and zona-free oocyteswere 37% (31/83) and 60% (49/81) respectively (P<0.01), with12 and 28% oocytes respectively showing polyspermia (P<0.05).No sperm penetration occurred in zona-intact oocytes. In experimentII the zona pellucida of hamster oocytes was partially openedby macro-or micro-manipulation techniques before the oocyteswere coincubated with human spermatozoa as in experiment I.Nine of 156 (6%) macro-manipulated and 11 of 133 (8%) micro-manipulatedoocytes lysed following zona-opening. Of 147 remaining macro-manipulatedand 122 remaining micro-manipulated oocytes, 80 (55%) and 79(64%) respectively were penetrated (P>0.05), with 30 and37% respectively showing polyspermia (P>0.05). These resultsdemonstrate that macro-manipulation results in similar penetrationand polyspermia rates as compared to conventional micro-manipulation.     

Seminal plasma can be a predictive factor for male infertility

Seminal plasma from ejaculates of 10 healthy, fertile volunteersand 63 infertile males was analysed for superoxide dismutase(SOD) and xanthine oxidase (XO) activities using a chemiluminometer.There was no statistically significant difference in the activityof either enzyme between control and infertile populations (113±74 IU/ml for SOD and 1.17 ± 0.52 IU/l for XO) in samplesfrom normozoospermic ejaculates. Sperm progressive motilitywas positively correlated with SOD activity in seminal plasmaof corresponding ejaculates (P < 0.05) and negatively withXO activity (P < 0.001). An ‘oxido-sensitive’index was defined as the SOD/XO ratio and was found to be inverselyrelated to sperm progressive motility samples (P < 0.01).Analysing this index among all tested samples of semen includingthose with pathological spermiograms, as well as normospermic(N) samples we found statistically significant (elevated) differencesin oligoasthenoteratospermia (OAT) in comparison with N (P <0.05); OAT samples were also significantly different from oligospermic(O) and oligoteratospermic (OT) samples (P < 0.05). Thissuggests that the ‘oxido-sensitive’ index of seminalplasma may be a simple diagnostic factor, useful in the determinationof male infertility.     

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: 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.     

Endocrinology: Luteal phase oestradiol and progesterone levels are stronger predictors than follicular phase follicle stimulating hormone for the outcome of in-vitro fertilization treatment in women with tubal infertility

Basal follicle stimulating hormone (FSH) in a natural cycle,FSH on cycle days 3 and 10 in a domiphene citrate-stimulatedcycle and oestradiol and progesterone area under the curve (AUC)in the luteal phase of the ciomiphene citrate-stimulated cyclewere evaluated as hormonal predictors for the outcome of FVFtreatment in 53 normally cycling women with tubal infertility.The pregnant women had significantly fewer treatment cycles(P < 0.001) and needed fewer ampoules of gonadotrophins (P< 0.001). They also had more oocyte retrievals (P < 0.001),more oocytes per retrieval (P < 0.01), higher fertilizationrate (P < 0.001) and more replaced pre-embryos per replacement(P < 0.01) as compared with non-pregnant women. Significantdifferences were found in FSH concentrations on cycle days 3(P < 0.05) and 10 (P < 0.001) after domiphene citratestimulation and for oestradiol and progesterone AUC in the lutealphase (P < 0.001) between those women who became pregnantand those who did not become pregnant after IVF treatment Lutealoestradiol and progesterone had considerably stronger predictivevalue for the outcome of IVF treatment as compared to basalFSH and domiphene citrate challenge test.     

Deterioration of sperm quality in young healthy Belgian men

We have retrospectively analysed the sperm characteristics of416 consecutive healthy young men who presented themselves inthe past 19 years as candidate sperm donors. Ejaculate volumeincreased slightly (P = 0.067), and average sperm concentrationdecreased (P = 0.035) by 12.4xlO6ml over the observation period,so that sperm count per ejaculate remained unchanged (P = 0.91).In contrast, sperm morphology (r = –0.23, P < 0.0001),rapid progressive motility (r = –0.42, P < 0.0001)and total motility (r = –0.33, P < 0.0001) presentedan important and time-related decrease. When a quadratic modelwas used rather than a linear one to analyse the data on rapidprogressive motility, there appeared to have been no furtherdecline since 1990. The average proportion of spermatozoa withnormal morphology decreased from 39.2% in the period 1977–1980to 26.6% in 1990–1995 (P < 0.0001), and the mean percentageof spermatozoa with rapid progressive motility decreased from52.7 to 31.7% (P < 0.0001). The percentage of candidate donorswith sperm characteristics below the 5th percentile cut-offvalue of a normal fertile population increased from 13 to 54%during the observation period (P < 0.0001). Since the techniqueof semen analysis has remained essentially unchanged in-so-faras has been practically possible, as has the method of recruitmentof candidate sperm donors, the observed deterioration of spermcharacteristics is considered to reflect degeneration of spermproduction among men aged between 20 and 40 years.     

Studies on psychosomatic implications of infertility-effects of emotional stress on fertilization and implantation in in-vitro fertilization

A case report shows the long and stony path of a couple whohad to go through eight attempts at IVF in 4 years until thedesired child was achieved. The psychosomatic implications ofthe various ‘failures’ of treatment become clearerwhen compared with the results of a psycho-endocrinologicalquestionnaire completed by 551 patients and 115 controls since1982. This questionnaire contains among others the Giessen personalitytest for the image of self, mother and father, a somatic complaintsscore to evaluate the severity of autoiiomic disorders, itemsfor the presence and severity of possible gynaeco-endocrinologicalsymptoms and items to evaluate the reference persons and theirchanges during childhood. The following results could be obtained.In a comparison of the means of 58 different somatic complaints,patients rated significantly higher than controls (P = 0.002).Women with menstrual disturbances, menstrual pains, acne orunclear skin or dry skin also suffered more from other complaints.These symptoms were also correlated with a higher score fordepressive mood. Those women who were not brought up by bothparents during their childhood also scored significantly higher(P < 0.0001) in the somatic complaints score. In the Giessenpersonality test patients appeared significantly more compulsive(P < 0.002) and depressive (P = 0.003) than the controlsand identified themselves more with their mothers. From 60 patientsin our IVF programme, 19 had no fertilization of oocytes andscored significantly higher (P = 0.008) on the social potencyscale than did the patients with fertilized oocytes. Women withunsuccessful treatments in terms of pregnancy scored significantlyhigher in the social resonance and social potency tests. Weinterpret this exaggerated positive attitude as an attempt toovercome inner fears, doubts and ambivalences. These resultssupport the importance of psychosomatic aspects in infertilityand reassure us of the necessity of including counselling inany infertility treatment, including IVF.     

Effects of antimicrobial therapy on sperm-mucus interaction

Sperm-mucus interaction under in-vitro or in-vivo conditionsmight be affected by microorganisms colonizing the reproductivetract. In order to study the influence of antimicrobial therapy,an extensive microbial screening was performed including Chlamydiatrachomatis, Mycoplasma hominis, Ureaplasma urealyticum, Neisseriagonorrhoeae, a broad spectrum of potentially pathogenic aerobicand anaerobic bacteria, Trichomonas vaginalis, herpes simplexvirus and yeasts. One-hundred-and-six couples with a mean durationof infertility of 5.5 years (range 1–12 years) and withisolation of potentially pathogenic microorganisms in semensamples and/or cervical swabs were submitted to a prospectivepilot study. None of the patients displayed signs or symptomsof infection in the lower genital tract. Before and after specifictherapy, based on antimicrobial susceptibility testing, spermanalyses and in-vitro sperm penetration meter tests (SPMT) (Kremer)were performed. SPMT was evaluated with cervical mucus of patients'wives, collected after a standardized oral treatment with oestrogensand, additionally, in a crossed manner with cervical mucus andspermatozoa of fertile donors. The success of antimicrobialtherapy was controlled by repeating the same mkrobial screeningand was 96%. However, there was a marked change in the mkrobialpattern. A comparison of the results of sperm analyses beforeand after treatment revealed neither significant differencesfor sperm volume, sperm count, propulsive motility, morphology,vitality, pH, fructose concentration or number of round cells,nor was there a significant influence on the cervical indexand the number of leukocytes in cervical mucus. In the SPMTpenetration distance, sperm density, quality and duration ofmotility were not significantly influenced by antimicrobialmedication, but were improved in the group of patients withvery poor SPMT results before treatment. The overall pregnancyrate was low (17% after 6 months, 21% after 12 months). Theresults suggest that microbial colonization is of minor importancefor sperm-mucus interaction and that the benefits of antimicrobialtherapy in asymptomatic couples are limited.     

Andrology: Comparison between fluorescent peanut agglutinin lectin and GB24 antibody techniques for the assessment of acrosomal status

In order to compare fluorescent peanut (Arachis hypogaea) agglutininlectin and GB24 antibody (specific for the inner acrosomal membrane)techniques for the assessment of acrosome reaction, both methodswere applied on semen specimens obtained from patients undergoingin-vitro fertilization (IVF). The acrosome status was evaluatedafter a 4 h incubation in B2 medium with and without calciumionophore A23187. Results obtained with both techniques werecompared and studied as a function of IVF outcome. The percentageof spontaneous acrosome-reacted spermatozoa was higher whenassessed by lectin than by GB24 (19 ± 2% versus 11 ±1%; P < 0.001). The difference between the two methods [lectinsminus GB24) was significantly higher in abnormal than in normalspermatozoa (10 ± 2% versus 4 ± 2%; P < 0.05),but did not significantly correlate with the percentage of acrosomeswith abnormal morphology (r = 0.28; NS). When studied in relationto the IVF results, the response to A23187 was higher in successesthan in failures (45 ± 2% versus 34 ± 4%; P <0.05) but there was no significant difference between methods.Thus the assessment of acrosome reaction is strongly influencedby the method used, particularly in abnormal spermatozoa. Sincethe results obtained with lectins were higher in abnormal spermatozoa,GB24 seems to be more effective for assessment of true acrosomereaction.     

Semen parameters and sperm morphology in men in unexplained recurrent spontaneous abortion, before and during a 3 year follow-up period

To investigate the role of the ‘male factor’ inthe patho-genesis of recurrent spontaneous abortion (RSA), especiallysperm morphology abnormalities, 120 previously selected coupleswith unexplained RSA were studied for sperm parameters retrospectivelyand prospectively. The patients were subdivided into three subgroups,depending on their reproductive outcome during the 3 years offollow-up study: (i) 48 RSA couples who achieved a successfulpregnancy; (ii) 39 RSA couples who experienced further abortions;and (iii) 33 RSA couples who experienced infertility duringthe follow-up period. A semen analysis was performed twice atthe time of inclusion in the study, and twice again during the3 year follow-up period. No significant differences in semenparameters were observed between the RSA males and fertile controls.Instead, significant differences were observed between the groupof RSA couples who experienced infertility during the follow-upand the other two groups (RSA couples who achieved successfulpregnancy and RSA couples who experienced miscarriages and nolive birth during the follow-up) for sperm concentration (P< 0.01 and P < 0.01 respectively), sperm motility (P <0.01 and P < 0.01 respectively) and sperm morphology abnormalities(P < 0.01 and P < 0.01 respectively). Sperm morphologyabnormalities do not seem to be involved in determining RSA;instead, they are an aetiological factor in determining infertilityin patients, along with the other semen parameters, in the RSAcouple‘s subsequent reproductive life. Semen analysisis an important test in the clinical management of RSA couples.     

Andrology: Seminal volume and total sperm number trends in men attending subfertllity clinics in the Greater Athens area during the period 1977-1993

The trends for such Important parameters of male fertility asseminal volume and total sperm number were assessed in men livingpermanently in the Greater Athens area over a prolonged periodof time. To this end, the records of three andrological laboratoriesemploying the same method for semen evaluation were analysedretrospectively. Out of 23 850 men examined from 1977 to 1993(17 years) for couple subfertillty, a total of 2385 (10%) wereselected for evaluation by a randomization procedure. Analysisof the data included (i) estimation of mean seminal volume andtotal sperm number per year, (ii) assessment of percentage frequencydistribution for each seminal parameter and (iii) evaluationof seminal volume and total sperm number changes in relationto the year of observation and age of the subjects. A significantdecrease (P < 0.01) of total sperm number was observed overthe years with a mean (± SEM) of 154.3 ± 19.2x 10⁶ at the beginning (1977), dropping to 130.1 ± 13.3x 10⁶ in the final year (1993). Mean seminal volume was lowerin the final year of observation, but its difference from theinitial year value was not significant Frequency distributionanalysis showed a marked decline in the 240–400 x 10⁶sub-set of the range of sperm number values from 16.9 ±4.5% (1977) to 10.6 ± 1.6% in the final year (P <0.01). Multiple regression analysis of seminal volume, totalsperm number, age and year of assessment revealed a significantdecline of the two seminal parameters along the years of observation(P < 0.05 and P < 0.0001 respectively). Over the sameperiod, a marked deterioration of some air pollution indiceswas observed in that area. It is concluded that in this raciallyand ethnically homogeneous sample of men, living under the sameenvironmental conditions, a significant decline in seminal volumeand total sperm number occurred over the 17 years of observation.