LH serum levels during ovarian stimulation as predictors of ovarian response and assisted reproduction outcome in down-regulated women stimulated with recombinant FSH

BACKGROUND: There has been much debate about the effect of 'residual' LH levels in normogonadotrophic women undergoing assisted reproduction with GnRH agonist down-regulation and recombinant FSH ovarian stimulation. The aim of this prospective study, where receiver-operating characteristic (ROC) analysis was used, was to assess further the usefulness of serum LH levels as predictors of ovarian response, assisted reproduction treatment outcome, and the outcome of pregnancy when measured throughout the ovarian stimulation period in a large cohort of such assisted reproduction treatment women. METHODS: A total of 246 consecutive women undergoing their first cycle of IVF or ICSI treatment were included in this study. Blood samples for hormone analyses were obtained on day S0 (the day when pituitary suppression was evidenced) and every other day from stimulation day 5 (S5) until the day of hCG injection. RESULTS: LH serum levels throughout ovarian stimulation treatment were similar for cancelled (n =32) versus non-cancelled (n = 214) cycles, non-conception (n = 132) versus conception (n = 82) cycles, and ongoing pregnancy (n = 66) versus early pregnancy loss (n = 16) groups. There was no correlation between LH serum levels in non-cancelled cycles and parameters of ovarian response and assisted reproduction treatment outcome. ROC analysis showed that serum LH concentration during ovarian stimulation was unable to discriminate between cancelled and non-cancelled cycles, conception versus non-conception cycles, or early pregnancy loss versus ongoing pregnancy groups. CONCLUSIONS: Serum LH measurements during ovarian stimulation with recombinant FSH under pituitary suppression in normogonadotrophic women undergoing assisted reproduction treatment cannot predict ovarian response, IVF/ICSI outcome, implantation, and the outcome of pregnancy. Thus, there is little underlying physiological support for the addition of LH in stimulation protocols if daily doses of an appropriate GnRH agonist (leuprolide or triptorelin having lower potency than buserelin) and a step-down regimen of recombinant FSH administration are used.     

The prognostic value of basal luteinizing hormone:follicle-stimulating hormone ratio in the treatment of patients with polycystic ovarian syndrome by assisted reproduction techniques

One of the main endocrinological disturbances in patients withpolycystic ovarian syndrome (PCOS) is the increased baselineconcentrations of luteinizing hormone (LH) and consequentlya high LHrfollicle-stimulating hormone (FSH) ratio. The aimof this study was to assess the relationship between the baselineLH:FSH ratio with the stimulation response and the miscarriagerisk in PCOS women stimulated for assisted reproduction techniques(ART) with and without gonadotrophin-releasing hormone analogue(GnRHa). Two groups of PCOS patients were analysed retrospectively.Group A (n = 20, 20 cycles) consisted of women stimulated withhuman menopausal gonadotrophin (HMG), and group B (n = 128,162 cycles) comprised women stimulated with buserelin-long/HMG.LH and FSH concentrations were measured during the early follicularphase (days 4–6) in a preceding spontaneous or progestininducedcycle. The following parameters were assessed: number of folliclesdeveloped, number of oocytes obtained and percentage of matureoocytes, as well as number of abortions and live births. Ingroup A, the baseline LH:FSH ratio was correlated inverselywith the number of follicles developed (P < 0.05), the numberof oocytes obtained (P < 0.05) and the percentage of matureoocytes (P < 0.05). In group B, no correlation was foundbetween the LH:FSH ratio and the number of follicles and oocytes,because their numbers were relatively constant irrespectiveof the baseline LH:FSH ratio, but a significant inverse correlationwas noted with the percentage of mature oocytes (P < 0.001).However, a comparison of the slopes of the curve indicated abetter correlation between the LH:FSH ratio and the percentageof mature oocytes in group A than in group B (P < 0.05).These findings were also confirmed when patients were subdividedaccording to the LH:FSH ratio (<3 or 3=3). Furthermore, inwomen who miscarried, the mean LH:FSH ratio was significantlyhigher than in women having a live birth. In conclusion, inPCOS patients stimulated with HMG, a high basal LH:FSH ratioappears to have an adverse effect on the number of folliclesand oocytes, as well as on oocyte maturity. On the other hand,the administration of GnRHa in the long protocol seems to reversethis detrimental effect on follicle and oocyte development.Furthermore, a higher LH:FSH ratio seems to predict a greaterpossibility for miscarriage, despite the use of GnRHa.     

Suppression of LH during ovarian stimulation: analysing threshold values and effects on ovarian response and the outcome of assisted reproduction in down-regulated women stimulated with recombinant FSH

BACKGROUND: It has been recently suggested that gonadotrophin-releasing hormone agonist down-regulation in some normogonadotrophic women may result in profound suppression of LH concentrations, impairing adequate oestradiol synthesis and IVF and pregnancy outcome. The aims of this study, where receiver-operating characteristic (ROC) analysis was used, were: (i) to assess the usefulness of serum LH measurement on stimulation day 7 (S7) as a predictor of ovarian response, IVF outcome, implantation, and the outcome of pregnancy in patients treated with recombinant FSH under pituitary suppression; and (ii) to define the best threshold value, if any, to discriminate between women with 'low' or 'normal' LH concentrations. METHODS: A total of 144 infertile women undergoing IVF/intracytoplasmic sperm injection (ICSI) treatment were included. Seventy-two consecutive patients having a positive pregnancy test (including 58 ongoing pregnancies and 14 early pregnancy losses) were initially selected. As a control non-pregnant group, the next non-conception IVF/ICSI cycle after each conceptual cycle in our assisted reproduction programme was used. RESULTS: The median and range of LH values in non-conception cycles, conception cycles, ongoing pregnancies, and early pregnancy losses, clearly overlapped. ROC analysis showed that serum LH concentration on S7 was unable to discriminate between conception and non-conception cycles (AUC(ROC) = 0.52; 95% CI: 0.44 to 0.61) or ongoing pregnancy versus early pregnancy loss groups (AUC(ROC) = 0.59; 95% CI: 0.46 to 0.70). To assess further the potential impact of suppressed concentrations of circulating LH during ovarian stimulation on the outcome of IVF/ICSI treatment, the three threshold values of mid-follicular serum LH proposed in the literature (<1, < or =0.7, <0.5 IU/l) to discriminate between women with 'low' or 'normal' LH were applied to our study population. No significant differences were found with respect to ovarian response, IVF/ICSI outcome, implantation, and the outcome of pregnancy between 'low' and 'normal' S7 LH women as defined by those threshold values. CONCLUSIONS: Our results do not support the need for additional exogenous LH supplementation in down-regulated women receiving a recombinant FSH-only preparation.     

Ovarian response and pregnancy outcome related to mid-follicular LH levels in women undergoing assisted reproduction with GnRH agonist down-regulation and recombinant FSH stimulation

BACKGROUND: The effect of LH levels on stimulation day 8 on ovarian response and pregnancy outcome were evaluated in women receiving pituitary down-regulation with GnRH agonists (0.8 mg Suprefact) s.c. daily until pituitary down-regulation and 0.4 mg/day during ovarian stimulation) and ovarian stimulation with recombinant FSH. METHODS: Blood samples were prospectively collected from a total of 207 normal women undergoing assisted reproduction and analysed retrospectively. Based on LH levels on stimulation day 8 patients were divided into four groups: <0.5, 0.51-1.0, 1.01-1.5, >1.51 IU/l. RESULTS: Estradiol levels on day 8 and estradiol per oocyte retrieved showed a highly significant correlation with LH concentrations on day 8. The total consumption of exogenous FSH and duration of gonadotrophin stimulation was inversely related to LH levels on day 8 (P < 0.002). The frequencies of fertilization and clinical pregnancies were superior in the two middle groups. Only 12% of the patients showed LH levels <0.5 IU/l, which, however, may be explained by the particular mode and doses of GnRH agonist used. CONCLUSIONS: Circulating levels of LH on day 8 have a significant impact on ovarian response and pregnancy outcome. LH should neither be too high nor too low. The mode of administration and the dose of the GnRH agonist used may determine the number of women who experience LH levels below 0.5 IU/l.     

Hypersecretion of luteinizing hormone and ovarian steroids in women with recurrent early miscarriage

Polycystic ovary syndrome is associated with hypersecretionof luteinizing hormone (LH) which has been implicated in theaetiology of early pregnancy loss. Although 82% of women withrecurrent early loss have polycystic ovaries on ultrasound imaging,random serum LH concentrations are normal. In the present study,we have obtained further information from serial samples concerningthe cyclical patterns of gonadotrophin and sex steroid secretionin these women. Twenty-one women with recurrent early pregnancyloss and 10 multiparous controls were investigated; 81% of themand one of ten control subjects had polycystic ovaries. Meanmid-follicular and mid-luteal serum LH and follicle stimulatinghormone (FSH) levels were similar in both groups. Seventeenwomen with pregnancy loss had either raised urinary LH excretionor a premature LH surge; one control subject had a prematureLH surge. Total LH excretion during the cycle and mean follicularphase serum testosterone was significantly greater with earlypregnancy loss than in the control group, the difference inLH being greatest in the early luteal phase. Urinary oestrone-3-glucuronideexcretion was raised in the early luteal phase of the cyclein the group with early pregnancy loss; there was no differencebetween the groups in pregnanediol-3-glucuronide excretion.These data demonstrate abnormalities in LH secretion in 81%of women with recurrent fetal loss. Inappropriately raised LHlevels may have adverse effects on the developing oocyte orendometrium either directly, or indirectly by causing an elevationin testosterone and oestrogen levels.     

Assessment of serum luteinizing hormone during ovarian stimulation with gonadortrophins

An immunoradiometrte assay (IRMA), using monoclonal antibodieswith high affinity for human luteinizing hormone (HLH), wasevaluated for quantitative measurement of serum LH after humanchorionic gonadotrophin (HCG) administration in patients undergoingstimulation of multiple folh'cular development. Compared toa radioimmunoassay (RIA) commonly used to monitor serum LH,LH IRMA was more effective by several orders of magnitude indiscriminating between HLH and HCG and showed no crossreactivityat HCG concentrations normally found in serum after hormonetreatment. Assays of serum samples obtained from 10 patientsreceiving HCG as part of an HMG/HCG protocol to induce ovulationfor IVF/GIFT also demonstrated that RIA values were greatlyaffected by exogenous HCG. It was estimated that 17–32%of serum HCG was measured as serum LH in RIA. In contrast, determinationsof serum LH by IRMA was not biased by exogenous HCG. Data fromIRMA indicated that eight of the 10 patients showed a significantrise in LH secretion, relative to mean baselines, at either12 or 36 h after adminstration. In one patient the rise hadalready occurred before HCG administration. When an LH riseoccurred, either before or after HCG injection, mean valueswere 2to 9fold higher than those of baseline levels. Assumingthat LH rises > 12 mlU/ml may relate to an endogenous surgeof LH, none of the patients showed a surge prior to HCG administration.On the contrary, the occurrence of an ‘LH surge’after HCG was apparent in four patients. These data demonstratethe application of monoclonal antibodies incorporated in anIRMA to study the occurrence of endogenous LH surges duringstimulation of follicular development by gonadotrophins.     

Pregnancy outcome following exposure to gonadotrophin-releasing hormone analogue during early pregnancy: comparisons in patients with normal or elevated luteinizing hormone

The outcomes of established pregnancies following the treatmentof infertile women with pituitary down-regulation before andduring treatment with ovulation induction and either intrauterineinsemination or timed intercourse were reviewed. Once startedon gonadotrophin-releasing hormone analogue (GnRHa) treatment,the patients were maintained on GnRHa therapy throughout thefollowing luteal phase to facilitate the start of the next treatmentcycle if no pregnancy was established. This resulted in patientstaking GnRHa until a positive pregnancy test indicated cessationof the treatment. The aim of our study was to determine whetherexposure to GnRHa during early pregnancy constituted a risk.Patients who were diagnosed as having elevated follicular phaseluteinizing hormone (LH) concentrations during their investigationswere analysed as a separate cohort to assess whether this diagnosishad implications with respect to pregnancy outcome. Out of 226recorded clinical pregnancies, 173 were traced and the datacollated: 16 cases resulted in clinical abortions, two wereectopic pregnancies and 155 women had live births at variousages of gestation. There were three pregnancies which were complicatedby congenital abnormalities. Patients with elevated LH concentrationson examination showed a higher rate of total pregnancy lossthan those with normal LH concentrations, despite the fact thatthe LH was suppressed during the cycle in which they conceived.The results suggest that pregnancy outcome is not adverselyaffected by GnRHa administration during the luteal phase ofthe conception cycle, and that the group diagnosed as havingelevated LH concentrations may retain their propensity to higherrates of pregnancy loss even when their LH concentrations aresuppressed during treatment.     

IVF/ICSI outcome and serum LH concentration on day 1 of ovarian stimulation with recombinant FSH under pituitary suppression

BACKGROUND: Down-regulation with GnRH agonist has been suggested to result in a profound suppression of LH bioactivity, reduced estradiol synthesis, and thus impaired IVF and pregnancy outcome. The aims of this study were: (i) to assess the usefulness of serum LH measurement on stimulation day 1 as a predictor of ovarian response, conception and pregnancy outcome in patients treated with long-term down-regulation with GnRH agonist and recombinant FSH, and (ii) to define the best threshold LH value, if any, to discriminate between women with different outcomes of IVF. METHODS: Records of 2625 cycles in 1652 infertile women undergoing IVF (n = 1856) and/or ICSI (n = 769) treatment were reviewed. RESULTS: The range of LH concentrations on stimulation day 1 overlapped among non-conception cycles, conception cycles, ongoing pregnancies and early pregnancy losses. Receiver operating characteristic (ROC) analysis showed that serum LH concentrations on stimulation day 1 were unable to discriminate between conception and non-conception cycles (AUC(ROC) = 0.51; 95% CI: 0.49-0.54) or ongoing pregnancies versus early pregnancy loss groups (AUC(ROC) = 0.52; 95% CI: 0.47-0.57). Stratification for various low serum levels of LH did not reveal significant differences with respect to conception or pregnancy outcome among different LH levels on stimulation day 1. CONCLUSIONS: Serum LH concentration on stimulation day 1 cannot predict ovarian response, conception and pregnancy outcome in women receiving long-term down-regulation during assisted reproduction treatment.     

Life table (survival) analysis to generate cumulative pregnancy rates in assisted reproduction: an alternative method of calculating the cumulative pregnancy rate in assisted reproduction technology

The current method of calculating cumulative pregnancy rate can lead to an overestimation of treatment efficacy, especially over many cycles of assisted reproduction treatment. The choice of scale of passage of time should be dependent upon the types of treatment to be evaluated. The number of treatment cycles to which patients' effort and commitment is directly related may be appropriate where the chance of pregnancy is expected to be significantly higher than non-treatment for them. Limiting the calculation of cumulative pregnancy rate only to the second or third cycle within 1 or 2 years will ensure that most patients are included in the calculation. More research is needed to assess different methods and develop better variables for assessing the efficacy of infertility treatment that can be informative for patients over the course of their treatment.     

Cumulative pregnancy and live birth rates in women with antiphospholipid antibodies undergoing assisted reproduction.

The aims of this study were to investigate the influence of antiphospholipid antibodies (APA) on cumulative pregnancy and live-birth rates in patients undergoing assisted reproductive treatment. Serum samples from 173 patients were collected prior to initiation treatment cycle and tested by enzyme-linked immunosorbent assay (ELISA) for the presence of immunoglobulin (Ig)G, IgM and IgA against cardiolipin, phosphoserine, phosphoethanolamine, phosphoinositol, phosphatidic acid, and phosphoglycerol. Fifty-six samples from patients who had at least two failed cycles by assisted reproductive treatment were also tested by a bioassay for the presence of lupus anticoagulants. Both cumulative pregnancy and live birth rates were not affected by the presence of any specific or any number of seropositive APA. There was no association between multiple assisted reproductive treatment failures and APA seropositivity. Neither the serum concentration of any of the 18 APA, nor the number of positive APA was correlated with the number of assisted reproductive treatment failed cycles or affected the probability of pregnancy. No patient was found to be positive for lupus anticoagulants. Using life table analyses, which has been recognized as the most appropriate method available to analyse assisted reproductive treatment results, we conclude that there is no relationship between circulating APA and assisted reproductive treatment outcome. APA do not affect the early process of implantation or maintenance of pregnancy among assisted reproductive treatment patients.     

The value of serum levels of oestradiol, progesterone and beta-human chorionic gonadotrophin in the prediction of early pregnancy loss.

Serial serum levels of oestradiol, progesterone and the beta-subunit of human chorionic gonadotrophin (beta-HCG) had been performed in 674 cycles in women conceiving a singleton pregnancy, either spontaneously or as a result of assisted conception. To determine the value of these estimations in the prediction of early pregnancy loss, frequency distribution curves and receiver operating characteristic curves were derived for the respective hormones measured at weeks 4-7 of gestation and expressed as multiples of the median (MoM) values in pregnancies occurring both with and without ovarian stimulation. A cut-off level of beta-HCG less than 0.5 MoM gave a sensitivity of 68% with an odds ratio of 4.0 at 7 weeks in unstimulated cycles in the prediction of pregnancy failure. A cut-off of 0.8 MoM for progesterone gave a sensitivity of 59% and an odds ratio of 2.8. Prospective hormonal monitoring during the early weeks of gestation may be useful in the prediction of early pregnancy loss and should help to avoid the emergency presentation of some of the complications of early pregnancy, in particular ectopic pregnancy. The limitations imposed by multiple pregnancies and uncertain gestation due to menstrual data may restrict the use of this strategy to specialist fertility centres.     

Increased early pregnancy loss in IVF patients with severe ovarian hyperstimulation syndrome.

BACKGROUND: Since severe ovarian hyperstimulation syndrome (OHSS) is a potentially life-threatening complication of assisted reproduction, the focus of attention in such cases is placed firmly upon the health of the patient, with the endeavour to achieve a pregnancy being considered of secondary importance. The aim of this study was to focus on the pregnancy rate and pregnancy outcome in IVF patients hospitalized for severe or critical OHSS, in one centre, during a period of 6 years. METHODS: We compared the characteristics of patients with severe OHSS: those who conceived with the ones who did not conceive, and among pregnant IVF patients, those with ongoing pregnancies with those that miscarried. RESULTS: Pregnancy was achieved in 60 of 104 (58%) patients with severe OHSS. Pregnancy continued until delivery in 37 of these 60 patients (62%), whereas the remaining 23 (38%) aborted. The pregnancy and abortion rates in patients with severe OHSS were significantly higher than those of IVF patients without OHSS, during the same time period [23% (1138/4922) and 15% (169/1138) respectively, P < 0.001]. The mean duration of hospitalization for OHSS was significantly shorter in those who delivered compared with those who aborted (5.9 +/- 3.2 versus 10.5 +/- 9.6 days, P < 0.01) and in the non-pregnant patients compared with the pregnant patients (5.2 +/- 3.2 versus 7.6 +/- 6.6 days, P < 0.02). CONCLUSIONS: The clinical pregnancy rate of IVF patients with severe OHSS was significantly higher than that of patients without the syndrome. A longer stay in hospital-reflecting a more severe form of OHSS-was correlated with a higher frequency of abortions. OHSS, necessitating hospitalization, is a detrimental clinical situation not only for the mother but also for the developing pregnancy.     

Pregnancy: Serum concentrations of luteinizing hormone and progesterone in ovum recipients: their relationship with pregnancy and miscarriage

Our objective was to test the hypothesis that the associationbetween elevated luteinizing hormone (LH) concentrations andmiscarriage is mediated via an effect of LH on the maternalenvironment, rather than on the oocyte. The impact of maternalage, ovarian function, previous IVF attempts, therapeutic (buserelin)and hormonal (LH, oestradiol, progesterone) effects occurringon the day of zygote intra-Fallopian transfer (ZIFT) or embryotransfer, and of oocyte or embryo numbers, whether they werefresh or frozen, and their mode of transfer on the occurrenceof pregnancy and miscarriage following ovum donation (n = 57)were investigated. The cycles were divided by outcome into non-pregnant(n = 26), miscarriage (n = 19) and normal term pregnancy (n= 12). The circulating concentrations of LH were greater inmiscarriage cycles (P = 0.046) and cycles ending in pregnancy(P = 0.04) than in non-pregnant cycles, while the concentrationsof progesterone were greater in non-pregnant (P = 0.029) andmiscarriage (P = 0.015) cycles than in cycles ending in pregnancy.Frozen embryos were used more frequently in non-pregnant comparedto cycles ending in pregnancy (P = 0.016). Multiple regressionanalysis was used to investigate which factors are associatedwith miscarriage and identified progesterone concentrationsat the time of transfer as being the only significant variable(r = 0.48, F = 8.5, P = 0.007). The same method of analysiswas used to investigate which factors are associated with thefailure to conceive and identified previous IVF attempts (F= 5.8, P = 0.021), the presence of ovarian function (F = 5.7,P = 0.022), the use of frozen zygotes (F = 5.1, P = 0.029) andprogesterone concentrations (F = 5.9, P = 0.02), with an overallresult of r = 0.59, F = 5.2 and P = 0.002. In conclusion, highprogesterone concentrations were associated with the failureto conceive and miscarriage. In contrast, LH concentrationswere lower in women who failed to conceive but similar in pregnantwomen who did and did not miscarry. This suggests that the associationbetween elevated LH concentrations and infertility is via adirect effect of LH on the oocyte and an indirect effect, mediatedby elevated progesterone concentrations, on the endometrium.     

Implantation is apparently unaffected by the dopamine agonist Cabergoline when administered to prevent ovarian hyperstimulation syndrome in women undergoing assisted reproduction treatment: a pilot study

BACKGROUND: Ovarian hyperstimulation syndrome (OHSS) is a result of ovarian overexpression of vascular endothelial growth factor (VEGF) and its receptor 2 (VEGFR2). VEGF/VEGFR2 binding disrupts cellular junctions and increases vascular permeability (VP), a characteristic of OHSS, but enhances angiogenesis, which is a fundamental step in implantation. In animals, the dopamine agonist Cabergoline (Cb2) prevents VP without affecting angiogenesis. In humans, Cb2 averts OHSS, but a possible detrimental effect on angiogenesis and implantation has not been explored. A pilot study was designed to analyze whether or not Cb2 administration, as a procedure for preventing OHSS, affects the outcome of assisted reproduction treatment (ART). METHODS: A retrospective study with endpoints of implantation and ongoing/term pregnancy rates. Women (n = 35) at risk of OHSS (20-30 follicles developed and >20 oocytes collected) took a daily oral dose of 0.5 mg Cb2 for 8 days, beginning on the day of hCG. They were matched with controls treated during the same period and who were similar with respect to age, number and quality of the embryos replaced, embryonic stage at transfer and sperm quality. RESULTS: No difference was detected between the groups in fertilization, implantation or pregnancy rates. A total of 14 ongoing (beyond 32 weeks) or full term pregnancies were registered in each group. No major problem was detected during pregnancy or after delivery in any of these babies. CONCLUSIONS: Administration of Cb2 in order to prevent OHSS is safe and does not appear to affect ART outcome.     

Value of serum CA-125 concentrations as predictors of pregnancy in assisted reproduction cycles

The means by which endometrial receptivity influences conception rates in assisted reproductive technology (ART) is poorly understood. As the glycoprotein CA-125 is a product of human endometrium and is measurable in the peripheral circulation, it was investigated whether it might serve as an indicator of endometrial receptivity and predictor of pregnancy. In this prospective study, serum CA-125 concentrations of 75 ART cycles were measured 1 day before and on the day of human chorionic gonadotrophin (HCG) administration, and on the day of oocyte retrieval. These women did not have endometriosis and were induced by long-protocol gonadotrophin-releasing hormone analogue. Pregnancy was achieved in 35 (46.7%) cycles, but not in 40 cycles (53.3%). Serum CA-125 concentrations 1 day before and on the day of HCG administration and on the day of oocyte retrieval were significantly higher in cycles with pregnancy than in those without pregnancy (P < 0.05). It was noted that CA-125 concentrations on the day of oocyte retrieval were the best predictors of pregnancy, with concentrations >10 IU/ml having an accuracy of 86.6% for pregnancy. In conclusion, in intracytoplasmic sperm injection cycles, women with high serum CA-125 concentrations (>10 IU/ml) on the day of oocyte retrieval had very high pregnancy rates.     

Insulin-like growth factor binding proteins in serum and follicular fluid from women undergoing ovarian stimulation with and without growth hormone

The effects of supplementary growth hormone (GH) upon insulin-likegrowth factor binding proteins (IGFBP) in serum and ovarianfollicular fluid were investigated in women undergoing buserelin-humanmenopausal gonadotrophin (HMG) ovulation induction for in-vitrofertilization. IGFBPs were detected by Western ligand blotting(WLB) or radio-immunoassay (IGFBP-3) and were shown in the presentstudy to increase and decrease in patients with diseases ofGH excess or deficiency. In the women undergoing treatment forovulation induction, radioimmunoassay of IGFBP-3 gave resultswhich were consistent with the well-documented GH dependencyof this protein, increasing in the serum when GH was administeredat the same time as busereli-HMG. In contrast there were noconsistent patterns in the abundance of the IGFBPs in serumand follicular fluid examined by WLB whether or not the patientwas receiving GH, and the IGFBPs varied independently of theresult obtained by radioimmuno-assay. Other studies have shownthat during pregnancy a serum protease can dramatically decreasethe detection of the IGFBPs on the WLB. However, there was noevidence for a protease in the serum or follicular fluid fromthese non-pregnant women undergoing ovulation induction. Tissueavailability of IGFs is probably regulated by the BPs, so thatdata would suggest that in normal women, neither ovarian activitynor GH at pharmacological concentrations is the primary regulatorof the IGFBPs, which are likely to be regulated by some otherfactor(s), e.g. nutrition. These data may account for the lackof consistent clinical improvement in studies investigatingthe hypothesis that supplementary GH during ovulation inductionwith gonadotrophins would be beneficial.     

Kinetics of SRY gene appearance in maternal serum: detection by real time PCR in early pregnancy after assisted reproductive technique

BACKGROUND: Fetal DNA circulating in maternal serum offers a possibility for non-invasive prenatal diagnosis but its kinetics during very early pregnancy is still unclear. In order to clarify this point, the studies on the kinetics of fetal DNA appearance in maternal serum were conducted on patients undergoing assisted reproduction. METHODS: Using a quantitative real time PCR assay, the presence of SRY gene sequences was evaluated in the serum of patients at the onset of pregnancy. RESULTS: Twenty-seven patients were originally studied but first trimester abortion occurred in five cases. Among the 22 ongoing pregnancies, ten were found to bear at least one male fetus and all sera from these women gave positive results for SRY gene detection. The SRY gene was found to be detectable as soon as day 18 after embryo transfer in one case and it had been found in the other nine patients by day 37. CONCLUSIONS: Fetal DNA is found in maternal serum even before the fetal circulation is established, which is highly suggestive that it is released, at least in part, from the trophoblast. Detection of fetal DNA in maternal serum very early in pregnancy may have clinical implications such as with the management of pregnant women carrying a fetus at risk for congenital adrenal hyperplasia.     

Comparison of the GnRH agonist and antagonist protocol on the same patients in assisted reproduction during controlled ovarian stimulation cycles

Despite the fact that both gonadotropin-releasing hormone (GnRH) agonist and antagonist protocol are effective in suppressing the incidence of premature luteinizing hormone (LH) surges through reversibly blocking the secretion of pituitary gonadotropins, the exact impact of these two distinctive protocols on the clinical setting of patients for in vitro fertilization and embryo transfer (IVF-ET) treatment, however, remained controversial. We thus in the present report conducted a retrospective study to compare the impact of GnRH agonist and antagonist protocol on the same patients during controlled ovarian stimulation cycles. A total of 81 patients undergoing 105 agonist and 88 antagonist protocol were analyzed. We failed to detect a significant difference between two protocols for the difference in duration of ovarian stimulation, number of recombinant FSH (Gonal-F) ampoules used, number of oocytes retrieved, serum levels for estradiol (E2) and progestone (P), thickness of endometrium, and the zygote- and blastocyst-development rate. It is seemly that high quality embryo rate was higher in the antagonist protocol, but the data did not reach a statistical significance. Nevertheless, Implantation rate and clinical pregnancy rate were significantly higher in the antagonist protocol (10.64% and 30.26%, respectively) than that of the agonist protocol (5.26% and 15.82%, respectively). Our data also suggest that the GnRH antagonist protocol is likely to have the advantage for improving the outcome of pregnancy in those patients with a history of multiple failures for the IVF-ET treatment.     

Early pregnancy factor as a marker for the earliest stages of pregnancy in infertile women

Early pregnancy factor (EPF) is known to be detectable in seraof pregnant women within 24 to 48 h after conception. To examinethe incidence of early embryonic loss after hormonal treatmentand homologous artificial insemination, we screened the seraof our patients for the presence of EPF by means of a rosetteinhibition test. Our results show a considerably lower conceptionrate as described in appropriate studies on fertile women. Ifreduced sperm quality is the indication for insemination, theabortion rate in the first 2 weeks of gestation is significantlyincreased in relation to the rest of our patients. hufertilitybecause of an Impaired development of the embryo, especiallyif repeated EPF-positive cycles do not result in a pregnancy,may be due to chromosomal defects or malformations and a chromosomalanalysis is recommended. In patients in whom EPF is never foundto be positive, indicating a disorder of conception, we regardIVF/ET to be the treatment of choice.     

Maternal serum ghrelin levels in early IVF pregnancies: lack of prognostic value for viable pregnancy and altered post-prandial responses

BACKGROUND: Ghrelin is a pleiotropic hormone, involved in the control ofgrowth and metabolism, whose circulating levels fluctuate inrelation to food intake and body mass index. Ghrelin has beendetected in the decidualized endometrium, as well as in humanand rat placenta. METHODS: A total of 106 patients undergoing IVF procedures were prospectivelyrecruited. On Days 16 and 23 after oocyte retrieval, the patientswere subjected to blood sampling after overnight fasting, fordetermination of serum ghrelin, hCGβ and progesterone levels.In addition, ghrelin levels were assayed in these groups, 2h after ingestion of a fixed-calorie meal. RESULTS: The subjects were divided according to whether they achievedan ongoing pregnancy. On Days 16 and 23 after oocyte retrieval,pre-prandial serum ghrelin levels were not statistically different,although a general trend toward a decrease in circulating ghrelinby Day 23 was detected in pregnant groups. Although in non-conceivingsubjects, maternal ghrelin levels showed an expected 15% declineafter meal ingestion, such a post-prandial decrease was notstatistically significant in pregnant women, selectively onDay 16 after oocyte retrieval. CONCLUSIONS: Maternal ghrelin levels at early gestational age do not appearto pose diagnostic (as marker) or prognostic value for pregnancyoutcome in IVF procedures.