The effect of RU486 administered during the proliferative and secretory phase of the cycle on the bleeding pattern, hormonal parameters and the endometrium

Seventeen healthy women aged 24–45 years with regularmenstrual periods, proven fertility and not using steroidalcontraceptives or IUD were recruited for the study. The volunteerswere followed during one control, one treatment and one follow-upcycle. Daily morning urine samples were obtained during thecontrol and the treatment cycle. The samples were analysed withregard to pregnanediol glucuronide (P₂-G), oestrone glucuronide(E₁-G), oestradiol (E₂), progesterone (P₄), LH and creatinine.During the entire 3-month study the subjects kept a record ofuterine bleeding and side effects. The subjects received 50mg RU486 daily either on cycle days 7–10 (n = 7) or oncycle days 20–23 (n = 10). An endometrial biopsy was takenon cycle day 10 in the first group and on cycle days 21–28in the second group of patients. Treatment during the proliferativephase caused significant prolongation of the cycle length dueto a delay of the oestrogen and LH surge. However, once theoestrogen concentration started to increase, the remaining partof the cycle was normal. The length of the follow-up cycle wassimilar to that of the control cycle. The morphology of theendometrium did not differ from control samples taken from untreatedwomen at the same time of the cycle. All ovulating women (n= 9) treated in the mid-luteal phase started to bleed on the3rd to 4th day of the treatment. In four of these women thebleeding was scanty and followed by a menstrual-like bleedingat expected time, while in the remaining five volunteers thetreatment bleeding was heavier and not followed by a new bleedinguntil a month later. The duration of the secretory phase was16.5 ± 1.3 days in women with two bleeding episodes and11.8 ± 1.9 days in women with one bleeding episode (P< 0.05). The hormonal parameters were similar in both groupsup to the start of the treatment. In the patients with one bleedingepisode, the treatment was associated with a reduction in progesteroneconcentration, while in the patients with two bleeding episodesthe progesterone concentration remained elevated until the secondbleeding episode. Light microscopic examination of the endometriumrevealed unique changes in the endometrial morphology. The resultsindicate that RU486 acts mainly on the endometrium but a director indirect effect on the corpus luteum cannot be excluded.The age of the corpus luteum may be of importance for its susceptibilityto RU486 treatment.     

The expression of {alpha}v and {beta}3 integrin subunits in the normal human Fallopian tube epithelium suggests the occurrence of a tubal implantation window

The co-expression of ₁ß₁, ₄ß₁ and _vß₃integrins in the human endometrium coincides with the implantationwindow. The _vß₃ integrin is expressed in the apicalsurface of the luminal epithelium and may serve to anchor trophoblastcells in the adhesion phase of implantation. Using immunohistochemistry,we compared the expression of _v, ₁, ₄ and ß₃ integrinsubunits in samples of normal human Fallopian tube and endometriumobtained from five women in the non-receptive period (lutealphase days 2–4) and from another five women in the receptiveperiod (luteal phase days 6–8). The staining was quantifiedvisually on a scale of 0 to ++, according to the intensity anddensity of stained cells. The _v subunit is expressed in theFallopian tube epithelium during both periods in a pericellulardistribution. The ß₃ subunit is also expressed inthe same location, but it is up-regulated during the periodof endometrial receptivity. The other subunits are expressedin localizations which are not relevant to trophoblast adhesionand exhibit little or no difference in the level of expressionbetween the non-receptive and receptive periods. Based on theseresults we postulate that the expression of the ß₃subunit in the human tubal epithelium is under the same systemiccontrolling signals as in the endometrium and that the normaltubal epithelium may have an implantation window, at about thesame time as the endometrium, that affords the opportunity fortrophoblast attachment should a 5–7 day embryo be undulyretained in the tube.     

Identification of specific serum proteins synthesized de novo by monolayer cultures of glandular cells of gestational endometrium

Monolayer cell cultures (n = 3) of glandular epithelium of gestationalendometrium obtained from three apparently healthy women undergoingelective termination of pregnancy (7–9 weeks gestation)were established. De novo synthesis of eight serum proteins(albumin, alpha₁-antitrypsin, cerulo-plasmin, beta-lipoprotein,alpha₂-macroglobulin, fibronectin and complement factors C3and C4) was demonstrated by the incorporation of radiolabelledsubstrate ([³⁵-S]methionine) employing autoradiography (AR)in combination with crossed immunoelectrophoresis (XIE), referredto as ARXIE, and line immunoelectrophoresis (LIE), referredto as ARLIE. By contrast, there was no evidence for de novosynthesis of IgA, haptoglobin and orosomucoid. Our findingssuggest that the gestational endometrium may contribute to theproduction of several proteins considered to be synthesizedand secreted mainly by the liver and reticulo-endothelial system.The simple techniques used here to identify the de novo synthesisof human serum proteins could be applied to investigate proteinsynthesis by a wide range of tissues and cells     

Rearrangement and expression of {chi} light chain genes can occur without {micro} heavy chain expression during differentiation of pre-B cells

The kinetics of light (L) chain gene rearrangement and expressionon mRNA and protein level has been studied with four stromalcell/IL-7 reactive, long-term in vitro proliferating pre-B celllines and clones, two from fetal liver of normal mice and twofrom fetal liver of E_µH-bcl-2 transgenic (bcl-2-tg) mice.These pre-B cell lines and clones are DJ_H-rearranged on bothH chain alleles. Two of the clones harbor H chain rearrangementswhich do not allow the expression of V_HDJ_H rearranged H chaingenes as _µH chain proteins. Upon removal of IL-7 fromthe pre-B cell cultures all four cell lines rearrange V_H-DJ_Hand V_L-J_L gene segments, loose the surface expression of c-kit,CD43, and surrogate light chain, as well as the capacity tobe clonable on stromal cells in the presence of IL-7. Pre-Bcells from normal mice die by apoptosis during differentiation,while those from bcl-2-tg mice do not. All four lines and clonesexpress comparable levels of mRNA for _µH and _µLchains with the same time kinetics during 3 days of differentiation.However, only two of the four pre-B cell lines and clones express_µH chain protein, whereas all four pre-B cell lines andclones express _µL chain protein at comparable levels between2x10⁵ and 1.40x10⁶ _µL chain molecules per cell. Theseresults suggest that _µH chain expression is not mandatoryfor rearrangement and normal expression of _µL chain geneswhen pre-B cells differentiate to B cells.     

Pregnacny: The abortifacient effect of misoprostol in the second trimester. A randomized comparison with gemeprost in patients pre-treated with mifepristone (RU486)

The objective of this work was to study the abortifacient effectsof misoprostol, an orally active prostaglandin E₁ (PGE₁) analogue,in the second trimester. A randomized study of two prostaglandinregimens in women pre-treated with the antiprogesterone mifepristonewas carried out in the gynaecological wards of Aberdeen RoyalHospitals, NHS Trust, and included 60 women at 13–20 weeks‘gestation, in whom termination of pregnancy had been agreed.Following pre-treatment with mifepristone 600 mg women wererandomly allocated to one of two prostaglandin regimens whichstarted 36–48 h later. The first misoprostol 400 µgorally (up to three doses) followed by gemeprost vaginal pessary1 mg up to two doses. The second was gemeprost vaginal pessary1 mg up to five doses. The main outcome measures were successrate induction-to-abortion interval and side-effects. Therewere no significant differences between the two groups in anyof the main outcome measures. We conclude that misoprostol isa stable, cheap PGE₁ analogue with demonstrable efficacy andacceptable side-effects in the management of second trimesterabortion. Further work is needed to establish the optimum doseand regimen.     

Difference in signalling between various hormone therapies in endometrium, myometrium and upper part of the vagina

BACKGROUND: Combined hormone treatments in post-menopausal women have different clinical responses on uterus and vagina; therefore, we investigated differences in steroid signalling between various hormone therapies in these tissues. METHODS: A total of 30 post-menopausal women scheduled for hysterectomy were distributed into four subgroups: control-group (n = 9), Tibolone-group (n = 8); estradiol (E(2))-group (n = 7); E(2) + medroxyprogesterone acetate (MPA)-group (n = 6). Medication was administered orally every day for 21 days prior to removal of uterus and upper part of the vagina. Tissue RNA was isolated, and gene expression profiles were generated using GeneChip technology and analysed by cluster analysis and significance analysis of microarrays. Apoptosis and cell proliferation assays, as well as immunohistochemistry for hormone receptors were performed. RESULTS: 21-days of treatment with E(2), E(2) + MPA or tibolone imposes clear differential gene expression profiles on endometrium and myometrium. Treatment with E(2) only results in the most pronounced effect on gene expression (up to 1493 genes differentially expressed), proliferation and apoptosis. Tibolone, potentially metabolized to both estrogenic and progestagenic metabolites, shows some resemblance to E(2) signalling in the endometrium and, in contrast, shows significant resemblance to E(2) + MPA signalling in the myometrium. In the vagina the situation is entirely different; all three hormonal treatments result in regulation of a small number (4-73) of genes, in comparison to signalling in endometrium and myometrium. CONCLUSION: Endometrium and myometrium differentially respond to the hormone therapies and use completely different sets of genes to regulate similar biological processes, while in this experiment the upper part of the vagina is hardly hormone responsive.     

Hormonal characteristics of follicular fluid from women receiving either GnRH agonist or hCG for ovulation induction

BACKGROUND: A recent prospective randomized study from our groupcompared GnRH agonist (0.5 mg buserelin) and hCG (10 000 IU)for triggering of ovulation following a flexible antagonistprotocol. The agonist group showed a poor reproductive outcomedespite luteal phase support with progesterone and estradiol(E₂). In the present prospective observational study, the healthstatus of follicles from the above study was monitored by analysingthe hormonal content of frozen/thawed follicular fluid samples.The aim was to test whether the poor reproductive outcome couldbe related to a defective pre-ovulatory follicular maturationresulting in oocytes with a compromised developmental competence.METHODS: Hormone concentrations were measured in two individualfollicular fluid samples from each of 32 women receiving buserelinand 37 receiving hCG, thus representing a subset of the folliclesretrieved. RESULTS: Follicular fluid levels of LH in the agonistgroup as compared with the hCG group was 11.1 ± 0.5 versus3.6 ± 0.3 IU/l (mean ± SEM; P < 0.001); FSH,6.3 ± 0.6 versus 3.3 ± 0.2 IU/l (P < 0.001);hCG, not determined versus 139±8 IU/l; E₂, 1.9 ±0.2 versus 1.8 ± 0.2 µmol/l (P > 0.10); progesterone,70 ± 4 versus 93 ± 6 µmol/l (P < 0.001);inhibin-A, 36.9 ± 3.1 versus 37.1 ± 2.5 ng/ml(P > 0.10) and inhibin-B, 35.6 ± 2.8 versus 40.1 ±3.1 ng/ml (P > 0.10). Thus, pronounced hormonal differencesexist in follicular fluid, and the collective concentrationof all three gonadotropins and the follicular fluid concentrationof progesterone were much higher in the group of women receivinghCG for ovulation induction. CONCLUSION: The study suggeststhat GnRH agonist results in proper pre-ovulatory follicularmaturation, but the ovulatory signal – probably in synergywith the resulting pituitary down-regulation – is toolow to support appropriate corpus luteum (CL) function.     

Two-dimensional gel analysis of human endometrial proteins: cyclic changes in the expression of specific proteins during the normal menstrual cycle

High resolution two-dimensional (2-D) gel electrophoresis wasused to compare the patterns of [³⁵S]methionine-labelled cellularproteins in endometrial tissue from healthy, normally menstruatingwomen. Samples of endometrial tissue were incubated with [³⁵S]methioninefor 20 h, and total cell lysates were processed for 2-D gelelectrophoresis. Using this technique it was possible to studyproteins with iso-electric points (pI) ranging from 3.5 to 11and relative molecular weights (M_r) ranging from 10 000 to 300000 Da. The fluorograms were compared by computer-aided analysiswhereby a total of 1095 [³⁵S]-labelled proteins were resolvedon the iso-electric focusing gels (IEF, pI 3.5–7) and488 on the non-equilibrium pH gradient electrophoresis (NEPHGE)gels (pI 6.5–11). Of the proteins on the IEF gels, 125showed differential expression during the menstrual cycle. Ofthese, 36 were maximally expressed in proliferative phase endometrium,26 in the interval phase and 63 in secretory and/or late secretoryphase endometrium. Correspondingly, on the NEPHGE gels a totalof 61 proteins exhibited cyclical variation, of which 30 weremore prominent in proliferative phase, 13 in interval phaseand 18 in secretory phase endometrium. This study shows that2-D gel electrophoresis is eminently suited to the identificationof proteins whose expression varies in a cyclical manner duringthe menstrual cycle. Further investigations should be carriedout to isolate and characterize these proteins with the aimof establishing useful markers for specific endometrial phasesof the menstrual cycle.     

Endocrinology: Prostaglandin F2{alpha} stimulates release of insulin-like growth factor binding protein-3 from cultured human granulosa-luteal cells

Human ovarian follicular fluid contains a number of insulin-likegrowth factor binding proteins (IGFBP) of which IGFBP-3 is themost abundant. IGFBP-3 synthesis is growth hormone-regulated.We studied the effect of prostaglandin F₂ (PGF₂) on IGFBP-3secretion by cultured human granulosa-luteal cells from follicularaspirates of women participating in an in-vitro fertilizationprogramme. The IGFBP-3 concentration was measured using a specificmonoclonal immunofluorimetric assay. Contrary to a previousreport on unstimulated follicles, this study demonstrated apositive correlation between follicular fluid IGFBP-3 concentrationand follicular size. PGF2a was found to stimulate in a dose-dependentfashion the secretion of IGFBP-3. Significant (p < 0.05)effects were found at PGF₂ concentrations of 10^–8, 10^–7and 10^–6 M. Because IGFBP-3 inhibits progesterone productionstimulated by insulin-like growth factor (IGF)-I, the PGF₂-inducedstimulation of IGFBP-3 production may be one of the mechanismswhereby PGF₂ exerts its luteolytic effect via the IGF system     

Endocrinology: Randomized trial of misoprostol and cervagem in combination with a reduced dose of mifepristone for induction of abortion

Mifepristone (600 mg) in combination with a prostaglandin hasbeen demonstrated to be a safe, acceptable alternative to vacuumaspiration for induction of abortion in the first 9 weeks ofpregnancy. However, the efficacy and side-effects of differentprostaglandins used in combination with mifepristone have notbeen assessed in a randomized trial. In this study, 800 womenseeking an abortion at gestational age 63 days amenorrhoea wererandomized to receive either 0.5 mg gemeprost by vaginal pessary(group I) or 600 µg misoprostol (group II) by mouth –48h after taking 200 mg mifepristone by mouth. The side-effectsand number of complete abortions were used as measures of efficacy.There was no significant difference in the rate of completeabortion between group I [96.7%; 95% confidence interval (CI)94.9–98.5%, n = 391] and group II (94.6%; 95% CI 92.3–96.9,n = 386). It was not possible to assess the outcome with certaintyin the remaining 23 women. However, there were significantlymore ongoing pregnancies in the women who received misoprostolthan in those who received gemeprost (nine versus one, P <0.01) and in eight of these 10 women the gestation was >49days. Fewer women in group II required analgesia than in groupI (48 versus 60%, P < 0.001) although the number requestingopiate was similar in each group (6.9 versus 5.2%, P > 0.4).The incidence of nausea and vomiting after misoprostol (47.8and 21.9% respectively) was higher (P < 0.001) than aftergemeprost (33.9 and 12% respectively). The incidence of infectionand heavy bleeding was low in both groups (<2%) and onlyone woman required blood transfusion. We conclude that the recommendeddose of mifepristone and gemeprost can be reduced without impairingclinical efficacy in pregnancies up to 63 days amenorrhoea.Misoprostol is a safe alternative prostaglandin but has a higherincidence of ongoing pregnancies especially at gestation after49 days amenorrhoea.     

In-vitro development of in-vivo produced rhesus monkey morulae and blastocysts to hatched, attached, and post-attached blastocyst stages: morphology and early secretion of chorionic gonadotrophin

The earliest time of secretion of chorionic gonadotrophin (CG)by primate embryos and its role during preimplantation developmentand implantation are not clearly determined. We cultured in-vivofertilized/developed zona-intact, morphologically normal morulae(n = 11) and early blastocysts (n = 11), freshly recovered (bynon-surgical uterine flushing) on days 5 and 6 of pregnancy,respectively (day 0 = the day following LH surge), from non-superovulatednaturally bred rhesus monkeys (Macaca mulatta). Embryos werecultured for a minimum of 24 days in dishes containing 1 mlof CMRL-1066 supplemented with 20% bovine fetal serum in a humidifiedatmosphere of 5% CO₂ in air at 37°C. The culture mediumwas changed every 48 h. The percentage of hatched blastocysts,developed from morulae and early blastocysts, was 90.9; elapsedtimes (mean ± SEM) were 67.8 ± 4.4 h (morula)and 37.8 ± 3.6 h (blastocyst). The minimum number ofHoechst-stained cells/hatched blastocyst was 531. The mean diameter(± SEM) of cultured embryos increased from 180 µmat the beginning of culture to 374 ± 28 and 450 ±19 µm at the fully expanded and hatched blastocyst stages,respectively. Hatched blastocysts continued to expand (maximumdiameter: 1125 ± 25 µm); after an additional 94–96h they attached firmly to the serum-coated dishes and producedhighly proliferating multinucleate trophectodermal cells, extendingto a maximum diameter of 2–6 mm by 11–21 days ofculture. Biologically active CG in embryo-grown, serial spentmedia samples was measured in a mouse Leydig cell bioassay.The embryonic secretion of CG (ng/ml, mean ± SEM) commencedjust prior to hatching ( 0.014 ± 0.0), increased to 1.7± 0.5 after hatching but prior to attaching, and to 122.7± 45.5 by 5–11, 5108.7 ± 1706.0 by 10–17days, and decreased to 317.0 ± 201.4 by 16–40 daysin culture. These results show firstly that in-vivo producedrhesus monkey morulae and early blastocysts develop in vitroto hatched and attached blastocyst stages, exhibiting extensivetrophectodermal outgrowths. Secondly, the secretion of bioactiveCG commences from low levels during the pre-attaching blastocyststage, and increases exponentially after the attachment andtrophectodermal outgrowth of cultured embryos.     

The effect of RU486 on the gene expression profile in an endometrial explant model

Administration of RU486 in vivo during the receptive phase rapidly renders the endometrium non-receptive to the implanting embryo. In order to identify key pathways responsible for endometrial receptivity we have used cDNA arrays to monitor gene expression changes in short-term endometrial explants in response to RU486. Endometrial biopsies from five normal fertile women at mid-secretory phase were cultured in the presence of estradiol and progesterone with or without RU486 for 12 h. cDNA arrays were produced containing approximately 1000 sequence-verified clones which included genes known to be important in angiogenesis, apoptosis, cell signalling, extracellular matrix remodelling and cell cycle regulation. cDNA probes from the paired endometrial samples were hybridized to the arrays and hybridization signals were quantified. A total of 12 genes displayed significant changes in expression; six were up-regulated and six down-regulated following RU486 treatment. For five of these genes this is the first report suggesting that they are regulated by steroids in the endometrium. JAK1 and JNK1 were two of the genes shown by the arrays to be down-regulated in RU486-treated endometrial explants. This was confirmed by real time RT-PCR. JAK1 immunoreactivity was localized to both glandular epithelium and the stroma of normal endometrium and staining was much stronger in the luteal phase of the cycle. These results show that components of two important signalling pathways in endometrium-the JAK/STAT pathway, and the JNK pathway-are altered by RU486. Genes whose expression is controlled by these pathways are likely to be involved in the mechanism by which steroids render the endometrium receptive to the implanting embryo.     

Association of SHBG gene polymorphism with menarche

The age of menarche may be subject to hereditary influencesbut the specific determinants are unknown. Our aim was to investigatethe possible association of a functional (TAAAA)_n polymorphismin the promoter of the sex hormone-binding globulin (SHBG) genewith the timing of menarche. This polymorphism has been associatedwith polycystic ovary syndrome (PCOS) and is considered to contributeto SHBG levels. We studied 130 healthy normal-weight adolescentfemales from a closed community in North–Western Greece.Information on menarche was obtained through interviews. TheBMI was recorded. Genomic DNA was isolated from peripheral bloodleukocytes for genotyping the TAAAA repeat region. We subdividedour subjects into two groups based on median age of menarche:those with menarche <13 years and those with menarche 13years. Genotype analysis revealed six (TAAAA)_n alleles containing5–10 TAAAA repeats. The distribution of alleles was differentin the two groups. Girls with late menarche had more frequentlylonger TAAAA alleles (>8 repeats), while girls with early menarchehad shorter alleles at a greater frequency (P=0.048). The majorcontribution to early menarche was by the 6 TAAAA repeat allele.Furthermore, carriers of the longer allele genotypes had latermenarche (13.24±1.15 years) than those with shorter allelegenotypes (12.67±1.15, P=0.018). These findings provideevidence for a genetic contribution of SHBG gene to the ageof menarche.     

Gonadotrophin surge-attenuating factor bioactivity is present in follicular fluid from naturally cycling women

Rat pituitary monolayer bioassays were used to compare gonadotrophinsurge-attenuating factor (GnSAF) bioactivity in follicular fluidfrom 12 follicles in 10 spontaneously cycling women with thatin pooled follicular fluid from women undergoing ovulation induction.Expressed as ED₅₀s (µl follicular fluid/well producing50% of maximal effect), GnSAF bioactivity was detectable inall spontaneous follicular fluid samples (1.4–33.3 µl/well)and in follicular fluid from women undergoing ovulation induction(6.8 µl/well). This GnSAF bioactivity was unaffected bypre-incubation with an inhibin antibody. When the data weregrouped according to whether the recovered oocytes fertilizedin vitro or not, the fertilized group contained significantlygreater GnSAF bioactivity than the unfertilized group (5.3 ±1.1 and 14.1 ± 2.6 µl/well respectively, P <0.05). While both inhibin bioactivity (9.7 ± 1.4 and28.9 ± 12.1 µl/well) and immunoreactivity (36.8± 2.2 and 21.0 ± 3.0 and ng/ml) were also greater(P < 0.01) in the fertilized compared with the unfertilizedgroups respectively, there were no other significant differencesbetween the two groups. We conclude that GnSAF is found in follicularfluid from spontaneously cycling women, supporting in-vivo evidencefor the involvement of GnSAF in feedback control of the ovary-pituitaryaxis.     

Variants in the ACVR1 gene are associated with AMH levels in women with polycystic ovary syndrome

BACKGROUND: Polycystic ovaries display an increased number of pre-antraland antral follicles compared with normal ovaries, suggestingthat early and late follicle development are disturbed. Thepathophysiology of this process is poorly understood. Sincethe transforming growth factor β family members, anti-Müllerianhormone (AMH) and bone morphogenetic proteins (BMPs), inhibitFSH sensitivity, their signalling may contribute to the aberrantfollicle development in these women. Here, we investigated therole of ALK2, a type I receptor for AMH/BMP signalling, in PCOSusing a genetic approach. METHODS: Seven single nucleotide polymorphisms in the ACVR1 gene, encodingALK2, were genotyped in 359 PCOS patients and 30 normo-ovulatoryand 3543 population-based control women, and haplotypes weredetermined. Subsequently, the association of ACVR1 variantswith ovarian parameters and hormone levels was investigated. RESULTS: The polymorphisms rs1220134, rs10497189 and rs2033962 and theircorresponding haplotypes did not show different frequenciesfrom controls, but were associated with AMH levels in PCOS women(P = 0.001, P = 0.002 and P = 0.007, respectively). Adjustmentfor follicle number revealed that the association with AMH levelswas, in part, independent from follicle number, suggesting thatvariants in ACVR1 also influence AMH production per follicle. CONCLUSIONS: Genetic variation within ACVR1 is associated with AMH levelsand follicle number in PCOS women, suggesting that ALK2 signallingcontributes to the disturbed folliculogenesis in PCOS patients.     

Immunology: Antiphospholipid antibodies and {beta}2-glycoprotein-I in 500 women with recurrent miscarriage: results of a comprehensive screening approach

Five hundred consecutive women (median age 33 years; range 19–45)with a history of recurrent miscarriage (median 4; range 3–16)were screened for the presence of antiphospholipid antibodies(APA)-lupus anticoagulant (LA) and/or anticardiolipin antibodies(ACA). The prevalence of persistently positive tests for LAwas 9.6% and for immunoglobulin G (IgG) and immunoglobulin M(IgM) ACA was 3.3 and 2.2% respectively. Only seven women (1.4%)were LA and ACA positive. Repeat testing, after an intervalof at least 8 weeks, demonstrated that only 65.7% of LA positive,36.6% IgG ACA positive and 36.0% IgM ACA positive women on initialtesting had a second positive test result. The dilute Russell‘sviper venom time detected the LA significantly more often thaneither the activated partial thromboplastin time or the kaolinclotting time (P < 0.001). There was no difference in thegestation of previous miscarriages between APA positive andAPA negative women. There was no difference in the plasma ₂-glycoprotein-Iconcentrations between APA positive and APA negative women withmiscarriages and normal women. All women with a history of recurrentmiscarriage should be tested for the presence of both LA andACA. A second confirmatory test should be performed in thosewith an initial positive test result.     

Efficacy of the sperm survival test for the prediction of oocyte fertilization in culture

The present study was carried out to investigate the predictivevalue of the sperm survival test (SST) with respect to the fertilizationof oocytes in culture. In general, our laboratory uses a totalof 50 000–150 000 motile spermatozoa to inseminate eachoocyte. The remaining material is evaluated for motility beforeand after 24 h of incubation at 37°C in a 5% CO₂ atmosphere.A total of 250 oocytes from 50 cases (mean ± SD, 5.0± 2.4 oocytes per retrieval) were inseminated and thefinal rate of cleaved embryos obtained was 52.5%. The SST (%)was considered normal when the ratio (final density of progressingspermatozoa after 24 h x 100/initial density of progressingspermatozoa) was 50% or more. Any other result was consideredabnormal. Cases presenting one or more cleaved embryos (n =40) were separated from those in which no embryo formation occurred(n = 10) and the results were compared in terms of the respectivesperm survival rates over a period of 24 h: normal SST (oneor more cleaved embryos, 37; none, five), abnormal SST (oneor more cleaved embryos, three; none, five). The specificityof the SST was 0.92 and sensitivity 0.50, the predictive valueof the abnormal test was 0.62 and the predictive value of thenormal test 0.88. The efficacy of the test was estimated at0.71, which was better than the conventional parameters of spermanalysis. A receiver — operating characteristics curvefor SST confirmed that the test can be useful for the predictionof fertilizability of oocytes in the laboratory.