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.     

17-beta-estradiol in relation to age at menarche and adult obesity in premenopausal women

BACKGROUND: We hypothesize that premenopausal endogenous estradiol may beassociated with age at menarche and adult overweight and obesity,potentially contributing to breast cancer risk. METHODS: We assessed age at menarche by questionnaire among 204 healthyNorwegian women, aged 25–35 years. Measures of body compositionincluded body mass index (BMI, kg/m²), waist circumference (WC,cm), waist-to-hip ratio (WHR) and fat percentage dual energyX-ray absorptiometry, (DEXA). Daily salivary 17-β-estradiol(E₂) concentrations were collected throughout one entire menstrualcycle and assessed by radioimmunoassay (RIA). Linear regressionanalyses and linear mixed models for repeated measures wereused and potential confounding factors and effect modifierswere tested. RESULTS: Among women with an early age at menarche (12 years), the overallmean salivary E₂ concentration increased by 3.7 pmol/l (95%confidence interval, 1.8–5.7 pmol/l) with each 9.8 cm(1 SD) increase in WC, which represents a 20.7% change in themean for the total group. Among the same early maturers, a 1SD (0.06) change in WHR was directly associated with a 24.0%change in mean E₂ concentration for the total group. CONCLUSIONS: Our findings support the hypothesis that early age at menarche,together with adult overweight and obesity, result in high levelsof 17-β-estradiol throughout the menstrual cycle.     

Epidermal growth factor receptors in human corpora lutea during the menstrual cycle and pregnancy

We have demonstrated the presence of epidermal growth factor(EGF) and its receptors in human non-gestational corpora lutea.To determine further the characteristics of EGF receptor binding,we examined 30 human corpora lutea throughout the luteal phaseand during pregnancy. Scatchard plots of EGF binding in 29 ofthe 30 corpora lutea were curvilinear, suggesting negative co-operativity.The mean ± SE of the association constant K_a was (0.9± 0.2) x 10⁹ 1/mol, the dissociation constant K_d was(2.2 ± 0.3) x10^–9 mol/1 and the number of bindingsites (Rt) was (15.8 ± 2.1) x10^–19 mol/µgprotein for non-gestational corpora lutea. The K_d increasedsignificantly in late pregnancies compared to early pregnancies(P = < 0.005), while Rt was significantly higher in termpregnancies than in either early pregnancy (P < 0.01) orthe menstrual cycle (P < 0.001). Corpora lutea atretica (n= 2) and ovarian stroma (n = 6) did not show any EGF bindingactivity. Our findings demonstrate the presence of specificEGF receptors in human corpora lutea of both the menstrual cycleand pregnancy. The changes in EGF binding parameters in earlypregnancy suggest that there may be a relationship between therole of EGF and ovarian steroidogenesis.     

Andrology: Two functional assays of sperm responsiveness to progesterone and their predictive values in in-vitro fertilization

We have recently reported, in a small cohort of subjects, thatacrosome reaction (AR) and intracellular free calcium ([Ca²⁺]_i)increase in response to progesterone were significantly correlatedwith in-vitro fertilization (IVF) rate. In the present studywe extended these results to 90 subjects undergoing IVF. Weconfirm that both parameters were highly significantly correlatedwith the fertilization rate (P<0.001). In particular, significantlylower responses to progesterone were detected in subjects witha fertilization rate <50%, further enlightening the functionalsignificance of sperm responsiveness to progesterone with respectto the process of fertilization. Moreover, we report here thatboth tests are highly discriminant of fertilization success,with positive predictive values >90% for [Ca²⁺]_i values whichincrease by >1.2-fold and AR inducibility >7% (cutoffvalues). Conversely, AR following challenge with the calciumionophore A23187 was less significantly correlated with thepercentage fertilization rate (P<0.05), and showed lowerpredictive values than response to progesterone. All these tests([Ca²⁺]_i increase in response to progesterone, AR in responseto progesterone and to A23187) appear highly sensitive and moderatelyspecific The positive predictive value may rise to >95% whenthe combination of two tests ([Ca²⁺]_i and inducibility of ARin response to progesterone) is considered. No correlation withfertilization rate has been found for spontaneous AR or basal[Ca²⁺]_i. In conclusion, we propose that assessment of humansperm responsiveness to progesterone may be clinically usefulin predicting fertilizing ability in vitro.     

Ovarian response to stimulation of HIV-positive patients during IVF treatment: a matched, controlled study

BACKGROUND: Our aim was to compare the ovarian response of HIV-positiveand -negative patients during IVF. METHODS: Setting –HIV and IVF reference university hospital. Twenty-seven HIV-infectedpatients who had undergone IVF between March 2000 and March2005 were matched with 77 HIV-negative patients for age, aetiologyof infertility, whether it was primary or secondary infertility,duration of infertility, history of pelvic surgery and typeof pituitary inhibition. Outcome – poor responders weredefined using one of the following criteria: a cancelled cycle(for insufficient ovarian response), less than four mature follicles(16 mm), peak serum levels of E₂ lower than 1000 pg/ml. RESULTS:There were no differences between the two groups of patientsfor the matched criteria. The proportion of African women andof women with a history of pelvic inflammatory disease was significantlyhigher among HIV patients than among the control group. Withthe exception of a lower number of transferred embryos amongHIV-positive patients versus HIV-negative ones (1.3 versus 1.9;P = 0.035), there was no significant difference between thetwo groups of patients regarding ovarian response parameters.CONCLUSION: HIV-infected patients who are in good general conditionand who are matched to a control group present a similar ovarianresponse to stimulation, suggesting the existence of a similarovarian reserve.     

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     

Andrology: Semen parameters as predictors of in-vitro fertilization: the importance of strict criteria sperm morphology

This study evaluated 120 couples undergoing in-vitro fertilizationtreatment to determine which semen parameter(s) predicted fertilizationand whether there was any consistent relationship between strictcriteria and standard assessment of sperm morphology. Strictcriteria morphology was the only significant predictor of fertilization(P = 0.0006, r² = 0.09), with a sensitivity of 94% and a specificityof 40%. A 12% cut-off point presented a negative predictivevalue of 98% and a positive predictive value of 22%. The probabilityof satisfactory fertilization is 40% with morphology <4%,which increases to 97% with normal morphology (12%). The receiveroperating characteristic curve deviated significantly from thediagonal with a 76% area wider the curve, making this a superiorpredictive test. This was augmented by likelihood ratios (LR)of 8.25 (LR +) for results with <4% normal morphology and0.15 (LR-) for results with 12% normal morphology by strictcriteria. While there was some correlation between strict criteriaand standard assessment of morphology (r²=0.35), the formerexplained only 12% (r²=0.12) of the variability in the latter.This study concludes that strict criteria morphology predictsfertilization, while other semen parameters do not. A 12% cut-offpoint makes strict criteria morphology an excellent predictorof satisfactory fertilization, while a value <4% is a goodpredictor of poor fertilization.     

Endometriosis: Prevalence and anatomical distribution of endometriosis in women with selected gynaecological conditions: results from a multicentric Italian study

We have evaluated the prevalence of endometriosis in selectedgynaecological conditions requiring surgery. Eligible for thestudy were women with primary or secondary sterility, chronicpelvic pain, fibroids or benign ovarian cysts requiring laparoscopyor laparotomy consecutively observed during the study periodin 23 obstetrics and gynaecology departments in Italy betweenMay 1991 and July 1992. Women with a previous diagnosis of endometriosiswere specifically excluded. A total of 3684 subjects enteredthe study. Of these, 660 (mean age 31 years) were included forsterility, 409 (mean age 32) for chronic pelvic pain, 1880 (meanage 42) for fibroids and 735 (mean age 33) for benign ovariancysts. During the surgical procedure surgeons were asked toexamine the pelvis carefully to identify endometriosis. Outof the 660 women included for sterility, 195 [30%, 95% confidenceinterval (CI) 26–35] had endometriosis; the correspondingfigures were 185 out of 409 (45%, 95% CI 39–52) for pelvicpain, 219 out of 1880 (12%, 95% CI 10–14) for fibroidsand 257 out of 735 (35%, 95% CI 31–40) for ovarian cysts;these differences were significant (x²₃ heterogeneity, absenceversus presence = 323.9, P < 0.001). Among women with endometriosiswho entered the study for sterility, 51% were at stage 1, 22%at stage 2, 20% at stage 3 and 7% at stage 4. The correspondingfigures for pelvic pain and fibroids were largely similar: 37%,24%, 30%, 10% for women with pelvic pain, 36%, 11%, 45% and8% for those with fibroids, but among cases with ovarian cystsstage 3 was over-represented (62% of cases). The most commonsites of endometriosis were, in order of frequency, the ovariesconsidered together, the posterior cul de sac and uterosacralligaments. Endometriotic implants were more common on the uterosacralligaments and the posterior cul de sac among women with sterilityand pelvic pain than in those with fibroids and ovarian cysts.The frequency of endometriosis was not directly related to ageat surgery, but decreased with increasing parity in all thefour criteria for entry groups.     

Pregnancy: Adverse effect of a homogeneous hyperechogenic endometrial sonographic pattern, despite adequate endometrial thickness on pregnancy rates following in-vitro fertilization

We have previously presented data to show that in patients whohad in-vitro fertilization (IVF)—embryo transfer usingovarian stimulation involving the luteal phase leuprolide acetate—humanmenopausal gonadotrophin (HMG) regimen, poor pregnancy resultsensued if either the endometrial thickness was < 10 mm ora homogeneous hyperechogenic sonograpic pattern was presentimmediately prior to taking a human chorionic gonadotrophin(HCG) injection. There were only 15 cases with this hyperechogenictype endometrium (and no pregnancies). The purpose of the presentstudy was to evaluate the influence of a hyperechogenic endometriumwhen the endometrial thickess was 10 mm, in a more extensiveseries, in women having IVF—embryo transfer using thesame ovarian stimulation regimen. A total of 273 consecutivecycles, where endometrial thickness was 10 mm, were evaluated(not including the 85 cycles previously reported). Of 22 patientswith the hyperechogenic pattern, one achieved a chemical pregnancy(-HCG >500 mIU/ml) and none achieved clinical pregnancies(ultrasound confirmation). In contrast, 67 of 251 (26.7%) patientsconceived with other echo patterns (x² analysis = 5.9, df =1, P = 0.01). These data thus confirm, in a larger series, thenegative influence of this type of echo pattern on subsequentpregnancy rates following the luteal phase leuprolide acetate—HMGovarian stimulation regimen.     

Comparison of SUZI and ICSI for severe male factor

We compare the results of subzonal insemination (SUZI) and intracytoplasmicsperm injection (ICSI) carried out between February 1993 andend of August 1994. A total of 232 couples underwent 302 cyclesof micro-assisted fertilization (79 patients had SUZI for atotal of 93 cycles, 153 patients ICSI for a total of 209 cycles).The indications for treatment were obstructive azoospermia in35 cycles, ejaculatory failure with severely low sperm countin 7 cycles, and failure of fertilization in a previous IVFcycle or less than 10% of oocytes fertilized in 87 cycles. In173 cycles the indication for treatment was a poor semen parameter.Patients undergoing ICSI had significantly higher fertilizationrates [43 (728/1692) versus 22.3% (151/676), ² = 86.308, P <0.0001], better chances of embryo transfer [95 (199/209) versus73% (68/93), ² = 30.671, P < 0.001], and greater numbersof embryos transferred (2.4 ± 0.9 versus 1.6 ±1.2 F = 42, P < 0.0001) than patients who had SUZI. Eighteenpatients became pregnant following the SUZI procedure, a pregnancyrate of 19% per egg collection, compared with 28% for thosewho underwent the ICSI procedure, where 58 out of 209 becamepregnant. The pregnancy rate was similar in those who underwentembryo transfer, whether they had ICSI or SUZI (29.2 and 28.6%respectively). Overall, the pregnancy rate doubled with eachnumber of embryos transferred, so it was 8.9% when one embryowas transferred, which increased to 18.3% when two embryos weretransferred, and this rose to 37.7% when three embryos weretransferred. There was no significant difference in the pregnancywastage rate between SUZI and ICSI. None of the offspring fromeither SUZI or ICSI showed any evidence of fetal abnormalities.Pregnancy rate was negatively correlated, with sperm progressionbeing 36% (36/100) if progression was <2 and 19.8% (40/202)if it was 2 (² = 8.99, P < 0.002). ICSI therefore providesa higher number of embryos available for transfer and shouldbe the primary treatment for severe male factor infertility.     

Higher-dimensional separation principle for the analysis of relaxation oscillations in nonlinear systems: application to a model of HIV infection

Author to whom all correspondence should be addressed In this paper, geometric and singular perturbation argumentsare utilized to develop a separation condition for the identificationof limit cycles in higher-dimensional (n 4) dynamical systemscharacterized by highly diversified time responses, in whichthere exists an (n - 3)-dimensional subsystem which quicklyreaches a quasi-steady state. The condition, which has beenused up to now to analyze relaxation oscillation in slow-fastsystems, is extended to accommodate dynamical systems in whichmore state variables are involved in a special manner whichstill allows for the use of singular perturbation techniques.Application is then made to a model of human immunodeficiencyvirus infection in T helper (T_H) cell clones with limiting restingT_H cell supply.     

Intracellular calcium increase and acrosome reaction in response to progesterone in human spermatozoa are correlated with in-vitro fertilization

In this study we have investigated responsiveness to progesteronein spermatozoa from a group of unselected male partners of couplesundergoing in-vitro fertilization (IVF). We evaluated progesterone-stimulatedintracellular Ca²⁺ ([Ca²⁺]_i) and percentage increase in acrosomereaction in the same sperm sample used for oocyte inseminations.[Ca²⁺]_i was measured with a fluorimetric method, while the acrosomereaction was assessed using a fluorescent probe (fluoresceinisothiocyanate-labelled peanut lectin). The average percentage[Ca²⁺]_i as well as the rate of increase in the frequency ofacrosome reaction following progesterone challenge were significantlylower (P < 0.005) in the group of patients with a fertilizationrate <50%. In addition, significant correlations betweenthe fertilization rate and the progesterone-stimulated [Ca²⁺]_iand acrosome reaction increases (r = 0.78 and r = 0.79 respectively)were observed. Furthermore, in cases of fertilization failure,no increase of [Ca²⁺]_i or acrosome reaction was observed inresponse to progesterone with the exception of one case. Ourresults indicate that [Ca²⁺]_i and acrosome reaction increasesin response to progesterone can be of value in the predictionof sperm fertilizing ability. As the two parameters were significantlycorrelated to each other (r = 0.86), the two assays have similarIVF predictive value and might be used interchangeably as adiagnostic tool in the assignment of male patients to the differentkinds of assisted fertilization techniques.     

Ovary and Ovulation: Human follicular nitric oxide pathway: relationship to follicular size, oestradiol concentrations and ovarian blood flow

Nitric oxide (NO) has been suggested to be involved in ovarianphysiology. Our aim was to study follicular nitrite and nitrate(NO₃/NO₂) levels in women undergoing in-vitro fertilization(IVF), and to examine their relationship to follicular size,oestradiol concentrations, and ovarian artery and intra-ovarianblood flow as measured by Doppler ultrasound. A total of 15patients from the IVF programme of Hadassah University Hospital,Mt Scopus, Israel, participated in the study. Detailed transvaginalultrasonographic examination was performed before ovum collection,and ovarian artery and intra-ovarian blood flow were measured.While aspirating the fofficles, the content of two to four ofthe fofficles in each ovary was collected individually, thevolume of follicular fluid measured, and NO₃/NO₂ concentrationswere determined. A statistically significant positive correlationwas found between follicular fluid NO₃/NO₂ concentrations andfollicular volume (r = 0.76), as well as between NO₃/NO₂ concentrationsand oestradiol concentrations (r = 0.63). A statistically significantnegative correlation was found between follicular fluid NO₃/NO₂concentrations and ovarian flow parameters as well as betweenNO₃/NO₂ concentrations in fofficles containing 4–5 mland ovarian artery pulsatility index.     

Pregnancy: Determinants of reproductive prognosis after ectopic pregnancy

The reproductive prognosis of 115 women desiring pregnancy whounderwent surgery for ectopic pregnancy between 1985 and 1990at the Clinica Luigi Mangiagalli, was analysed after a medianfollow-up period of 26 months (range 2–83). Probabilityof reproductive events was assessed by a product-limit model.Women who underwent surgery for ectopic pregnancy had a 54%probability of becoming pregnant (cumulative pregnancy rate,CPR) and a 36% probability of giving birth to a child (cumulativelivebirth rate, CLB) during the 3 years after surgery. Thesepercentages dropped with history of previous ectopic pregnancy(respectively 33%, P = 0.07, and 7%, P < 0.05). Increasingage at surgery and presence of adhesions in the contra-lateraltube seemed to be associated with poor reproductive prognosis(CPR = 40% and CLB = 12% for women aged 35 years and CPR = 37%and CLB = 20% in women with adhesions in the contra-lateraltube), but these findings were not statistically significant.No association emerged between fertility and parity or typeof surgery. The recurrence rate of ectopic pregnancy was 20%.No significant association emerged between recurrence of ectopicpregnancy and age, history of previous pregnancy, history ofprevious ectopic pregnancy, non-intact contra-lateral tube andsalpingotomy.     

Skewed T cell receptor V{alpha} repertoire among superantigen reactive murine T cells

Reactivity of murine T cells with viral or bacterial superantigensis clearly correlated with the expression of TCR V_ßdomains. Thus, T cells responding to the minor lymphocyte stimulatorylocus (Mls-1^a) or staphylococcal enterotoxin B (SEB) expresspredominantly TCR V_ß6 or V_ß8.2 respectively.We have investigated the involvement of the other major variableelement of the TCR, the V domain, in these superantigen responses.Using a panel of anti-TCR V mAbs, It is demonstrated that theTCR V repertoire among superantigen stimulated V_ß6⁺or V_ß8.2⁺ blasts (responding to Mls-1^a or SEB respectivelyin vitro) is altered in comparison with anti-CD3 stimulatedcells expressing the same V domains. Furthermore, the TCR Vrepertoire is strongly skewed in TCR V_ß8.2 transgenicmice that have undergone extensive peripheral clonal deletionafter SEB injection. These data imply that the V domain influencessuperantigen recognition by sthe TCR.     

Anti-Mullerian hormone indicates early ovarian decline in fragile X mental retardation (FMR1) premutation carriers: a preliminary study

BACKGROUND: Women who carry the fragile X mental retardation (FMR1) premutationare at risk for fragile X-associated primary ovarian insufficiency.Past studies have shown that carriers who are still cyclinghave increased levels FSH compared with non-carriers. As anti-Mullerianhormone (AMH) has been shown as an excellent marker of ovariandecline, we examined AMH levels among premutation carriers tocharacterize their ovarian function. METHODS: We determined the level of FSH and AMH in serum samples collectedduring early follicular phase from women who carried longerFMR1 repeat alleles (defined as 70 repeats, n = 40) and thosewith shorter repeat alleles (<70 repeats, n = 75), identifiedby DNA analysis. Comparisons were made stratified by age andcarrier status. RESULTS: For all age groups, AMH levels were significantly lower amonglonger repeat allele carriers compared to shorter repeat allelecarriers (P = 0.002, 0.006 and 0.020 for women ages 18–30,31–40 and 41–50 years, respectively). In contrast,increased FSH indicative of early ovarian decline was only evidentfor longer repeat allele carriers aged 31–40 years (P= 0.089, 0.001 and 0.261 for women ages 18–30, 31–40and 41–50 years, respectively). CONCLUSIONS: These preliminary data suggest that AMH levels indicate earlyovarian decline among women with longer FMR1 repeat alleles;moreover, AMH appears to be a better marker than FSH in identifyingthis early decline.     

Effects of strontium on the permeation and gating phenotype of calcium channels in hair cells

To minimize the effects of Ca²⁺ buffering and signaling, this study sought to examine single Ca²⁺ channel properties using Sr²⁺ ions, which substitute well for Ca²⁺ but bind weakly to intracellular Ca²⁺ buffers. Two single-channel fluctuations were distinguished by their sensitivity to dihydropyridine agonist (L-type) and insensitivity toward dihydropyridine antagonist (non-L-type). The L- and non-L-type single channels were observed with single-channel conductances of 16 and 19 pS at 70 mM Sr²⁺ and 11 and 13 pS at 5 mM Sr²⁺, respectively. We obtained K_D estimates of 5.2 and 1.9 mM for Sr²⁺ for L- and non-L-type channels, respectively. At Ca²⁺ concentration of 2 mM, the single-channel conductances of Sr²⁺ for the L-type channel was 1.5 and 4.0 pS for the non-L-type channels. Thus the limits of single-channel microdomain at the membrane potential of a hair cell (e.g., –65 mV) for Sr²⁺ ranges from 800 to 2,000 ion/ms, assuming an E_Ca of 100 mV. The channels are 4-fold more sensitive at the physiological concentration ranges than at concentrations >10 mM. Additionally, the channels have the propensity to dwell in the closed state at high concentrations of Sr²⁺, which is reflected in the time constant of the first latency distributions. It is concluded that the concentration of the permeant ion modulates the gating of hair cell Ca²⁺ channels. Finally, the closed state/s that is/are altered by high concentrations of Sr²⁺ may represent divalent ion-dependent inactivation of the L-type channel.     

Immunology: Formation of antisperm antibodies in women treated with Fallopian tube sperm perfusion

Fallopian tube sperm perfusion (FSP) is a combination of ovarianstimulation and intra-uterine insemination using a large volume(4 ml) of inseminate containing 10⁷–10⁸ spermatozoa. Theinseminate will flush the Fallopian tubes and some of it willend up in the pouch of Douglas. In the present study, we haveinvestigated whether the FSP method will result in the formationof serum antisperm antibodies in the female. A total of 184treatment cycles were given to 128 women. The indications fortreatment were: unexplained infertility (n = 35), various infertilitydiagnoses (n = 28) and donor insemination (n = 65). Prior totreatment, 11 (8.6%) women had a positive tray-agglutinationtest (Friberg) and/or a positive immunobead test. After completingone to four treatment cycles, another six (4.7%) women had developedserum antisperm antibodies. The antibodies induced by the treatmentwere of isotype IgM and directed against the tailtip of thespermatozoa. Two of the women, who prior to the treatment hadantisperm antibodies, showed an increase in antibody titre duringtreatment. There was no statistically significant differencein the pregnancy rate between the women with antisperm antibodiesand the women without. In our opinion, the small risk of developingantisperm antibodies is no contra-indication for treating infertilecouples with FSP.     

Andrology: Strategies in frozen donor semen procreation

Data were analysed from 710 couples who had been assessed todetermine the effectiveness and the drawbacks of three differentmethods of insemination using frozen donor semen. Intracervicalinsemination (ICI) was the first method used when the womenhad no tubal disorder: 255 pregnancies were achieved in a totalof 2558 cycles (10%). Intrauterine insemination (IUI) associatedwith ovarian stimulation resulted in 152 pregnancies over 966cycles (16%). In-vitro fertilization (IVF) was proposed after12 insemination failures using either of the other methods orwhen the initial gynaecological examination had revealed abnormalitiessuch as tubal occlusions; 48 pregnancies were obtained in 262cycles (18.3%). The pregnancy rate using ICI was significantlyhigher when two inseminations were performed per cycle, comparedwith one insemination per cycle (12.3 versus 7%, P < 0.001).The number of motile spermatozoa per straw was correlated withthe pregnancy rate when using ICI, rising from 9% with <4X10⁶motile spermatozoa to 13.8% with 4–8X10⁶ and 17.2% with>8X10⁶. No relationship was found between the number of motilespermatozoa and the pregnancy rate using IUI and IVF. The incidenceof primary ovulatory disorder was higher among women whose husbandswere oligozoospermic than among those whose husbands were azoospermic(19 versus 9%, P < 0.01), but ovarian stimulation improvedthe fecundity of subfertile women. The outcome of pregnancieswas also analysed for the three methods. From these data, strategicplans have been proposed to maximize the pregnancy rate forwomen undergoing therapeutic donor insemination with frozensemen.     

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