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PubMed1998

Leukemia inhibitory factor in human reproduction.

PROBLEM: Leukemia inhibitory factor (LIF) is a pleiotropic cytokine of the interleukin-6 family and has different biological actions in various tissue systems. Although named for its ability to inhibit proliferation of a myeloid leukemic cell line by inducing differentiation, it also regulates the growth and differentiation of embryonic stem cells, primordial germ cells, peripheral neurons, osteoblasts, adipocytes, and endothelial cells. LIF is crucial for successful implantation of the embryo in mice. Currently, there is an accumulation of data about the role of LIF in human reproduction. METHOD OF STUDY: This review of the literature and of our studies focuses on the expression, regulation, and effects of LIF in the human endometrium, fallopian tube, and ovarian follicle. RESULTS: Human endometrium expresses LIF in a menstrual cycle-dependent manner. Maximal expression is observed between days 19 and 25 of the menstrual cycle, coinciding with the time of implantation. Various cytokines and growth factors induce endometrial LIF expression in vitro. LIF receptor is expressed in endometrial tissue throughout the menstrual cycle and on human blastocysts in a stage-dependent manner. Affecting the trophoblast differentiation pathway toward the adhesive phenotype, LIF plays a role in implantation. LIF is also expressed and secreted by the epithelial cells of the fallopian tube. Its increased expression in the tubal stromal cell cultures by the inflammatory cytokines suggests a link between salpingitis and ectopic implantation in the tube. The rising follicular fluid LIF level around the time of ovulation indicates that LIF may play a role in ovulatory events, early embryonic development, and implantation. CONCLUSIONS: There is growing evidence that LIF may be one of the entities that plays a role in human reproduction.

Animals↗

PubMed1988

Outcome of pregnancy in underweight women after spontaneous and induced ovulation.

Low maternal weight before pregnancy and poor weight gain during pregnancy are known to result in an increased prevalence of low birthweight infants. Low body weight is also an important cause of amenorrhoea. The hypothesis that amenorrhoeic underweight women who become pregnant after induction of ovulation are more at risk of delivering low birthweight infants than underweight women who ovulate spontaneously was investigated. Forty one pregnant women in whom ovulation had been induced and 1212 in whom ovulation was spontaneous were studied. Women ovulating spontaneously whose weight was normal and who showed good weight gain during pregnancy (greater than 450 g a week) had the lowest incidence (6%) of babies who were small for gestational age. Underweight women (body mass index less than 19.1) who ovulated spontaneously had a threefold increased risk of delivering babies who were small for gestational age (18%). Overall, the women in whom ovulation had been induced had an even higher risk of babies who were small for dates (25%), and the risk was greatest (54%) in those who were underweight. The outcome of pregnancy is related to weight before conception, which in many cases reflects nutritional state; lack of spontaneous ovulation indicates an increased risk of producing a small for dates infant. The most suitable treatment for infertility secondary to weight related amenorrhoea is therefore dietary rather than induction of ovulation.

Body Weight↗

PubMed1986

Sonographic evaluation of normal and induced ovulation.

The adult premenopausal ovary exists in a dynamic state of flux, with ovarian position, size, activity, and structure constantly undergoing changes. Pelvic sonograms must be interpreted with knowledge of the stage of the menstrual cycle and the expected normal range of sonographic findings, because features that are normal in one phase may be abnormal in another. Only against this background knowledge can the challenging task of interpretation of pelvic sonograms be successfully performed. US also plays an increasingly important role in monitoring ovulation induction and alternative methods of fertilization and has improved pregnancy rates by distinguishing between the presence of one or more mature follicles as opposed to a cohort of immature follicles, suggesting the possibility of multiple ovulation with the risks of hyperstimulation or poor pregnancy outcome from multiple gestation, indicating optimum timing of human chorionic gonadotropin administration, allowing detection and confirmation of normal follicular rupture and ovulation, indicating optimum timing of oocyte retrieval in programs of in vitro fertilization with embryo transfer (IVF-ET) or insemination programs, and assessing pathologic pelvic conditions and ovarian accessibility in IVF-ET programs. Oocyte retrieval and embryo transfer are also being performed under US guidance.

Anovulation↗

PubMed1980

Influence of alpha-melanotropin on LH release in the rat.

The effects of MSH on spontaneous and induced ovulation as well as on serum LH levels were studied. Rats which received progesterone on D1 and D3 were injected i.p. with 30 microgram of alpha-MSH or saline solution and examined for ovulation on the following day. The injection of alpha-MSH induced ovulation in 90% of the animals; saline solution, on the contrary, induced an ovulatory response only in 10% of the cases. Rats injected with beta-MSH yielded similar results to those injected with saline solution, indicating a possible specific alhpa-MSH action. The replacement of the second progesterone injection by alpha-MSH failed to increase to ovulation indicating that alpha-MSH synergizes or alters progesterone action. It would seem that alpha-MSH stimulates ovulation-potentiating progesterone effects on the CNS. LH effectively induced ovulation in rats where spontaneous ovulation was blocked by an injection of progesterone on D1. The i.p. injection of 30 microgram alpha-MSH lowered the amount of LH necessary to induce ovulation. Rats which received alpha-MSH and LRH showed a marked increase in serum LH levels compared with rats where alpha-MSH had been replaced by saline solution. Collectively the information provided in this paper is consistent with the view that the release of MSH observed in the morning of PE would signal the discharge of LH from the pituitary gland occurring in the afternoon of PE.

Animals↗

PubMed2004

Regulator of G-protein signaling-5 induction in pericytes coincides with active vessel remodeling during neovascularization.

We identified regulator of G-protein signaling-5 (RGS-5) as an angiogenic pericyte marker at sites of physiologic and pathologic angiogenesis. In a mouse model of pancreatic islet cell carcinogenesis, RGS-5 is specifically induced in the vasculature of premalignant lesions during the "angiogenic switch" and further elevated in tumor vessels. Similarly, RGS-5 is overexpressed in highly angiogenic astrocytomas but not in hypoxia-inducible factor-1alpha (HIF-1alpha)-deficient tumors, which grow along preexisting brain capillaries without inducing neovessels. Elevated levels of RGS-5 in pericytes are also observed during wound healing and ovulation indicating a strong correlation between RGS-5 expression and active vessel remodeling beyond tumor angiogenesis. Moreover, antitumor therapy, which reverses tumor vasculature to an almost normal morphology, results in down-regulation of RGS-5 transcription. Taken together, these data demonstrate for the first time a factor that is specific for "activated" pericytes. This further supports the notion that pericytes, like endothelial cells, undergo molecular changes during neovascularization that makes them a novel target for antiangiogenic therapy.

Animals↗

PubMed2003

Leptin and leptin receptor expression in the rat ovary.

Leptin is an important satiety hormone and reproductive regulator and is found, along with its receptors, throughout the ovary. To date, the changes in ovarian expression of both of these proteins throughout the estrous cycle has not been studied, and the examination of protein expression has not distinguished between different forms of the receptor. In this study leptin mRNA expression in the immature gonadotropin-primed rat ovary increased 3-fold after human chorionic gonadotropin administration, followed by a dramatic increase in mRNA for both the short form (Ob-Ra) and the long form (Ob-Rb) of the leptin receptor (approximately 8- and 7-fold, respectively). A corresponding increase in mRNA expression of the receptor was not observed in isolated preovulatory follicles. Using immunohistochemistry, we observed protein expression of the long form of the leptin receptor (Ob-Rb) in the ovary, with high intensities observed in oocytes and endothelial cells as well as thecal cells and corpora lutea. These results suggest that ovarian expression of leptin and its receptor is regulated across the cycle by gonadotropins, with peak expression at ovulation, indicating a possible involvement in oocyte maturation, angiogenesis, follicle rupture, or subsequent corpus luteum formation.

Animals↗

PubMed2004

Pharmacological inhibition of phosphodiesterase 4 triggers ovulation in follicle-stimulating hormone-primed rats.

Phosphodiesterases (PDEs) are a family of enzymes that hydrolyze cyclic nucleotides to render them biologically inactive. As such, these enzymes are critical regulators of signal transduction pathways that use cyclic nucleotides as second messengers. PDE4 is one such member that has been identified in ovarian tissue and purported to have a role in the regulation of gonadotropin action. In the present study, selective PDE4 inhibitors enhanced intracellular signaling in a human LH receptor-expressing granulosa cell line. In vivo, PDE4 inhibition in FSH-primed rats resulted in ovulation, indicating that the PDE4 inhibitors can substitute for LH and human chorionic gonadotropin (hCG) in this process. Moreover, when coadministered with a subeffective dose of hCG, PDE4 inhibitors acted synergistically to enhance the ovulation response. Inhibitors of PDE3 or PDE5 had no ovulatory effect under similar conditions. Oocytes that were ovulated after PDE4 inhibition could be fertilized in vitro at a rate similar to that of oocytes from hCG-induced ovulation. Moreover, such oocytes were fully capable of being fertilized in vivo and developing into normal live pups. These results indicate that small molecule PDE4 inhibitors may be orally active alternatives to hCG as part of a fertility treatment regimen.

3',5'-Cyclic-AMP Phosphodiesterases↗

PubMed2002

Caspase-3 is a pivotal mediator of apoptosis during regression of the ovarian corpus luteum.

Because caspase-3 is considered a primary executioner of apoptosis and has been implicated as a mediator of luteal regression, we hypothesized that corpora lutea (CL) derived from caspase-3 null mice would exhibit a delayed onset of apoptosis during luteal regression, when compared with CL derived from wild-type (WT) mice. To test this hypothesis, ovulation was synchronized in immature (postpartum d 24-27) WT and caspase-3-deficient female littermates by exogenous gonadotropins. Individual CL were isolated by manual dissection, 30 h after ovulation, and placed in organ culture dishes in the absence of serum and growth factors. At the time of isolation (0 h) and after 24, 48, and 72 h in culture, the CL were removed and assessed for the presence of processed (active) caspase-3 enzyme and for apoptosis by multiple criteria. There was no evidence of active caspase-3 enzyme or apoptosis in either WT or caspase-3-deficient CL before culture. However, CL derived from the WT mice exhibited a time-dependent increase in the level of active caspase-3 and apoptosis during culture. By comparison, CL derived from caspase-3-deficient mice, cultured in parallel, failed to exhibit any detectable active caspase-3 and showed attenuated rates of apoptosis. To extend these findings derived from ex vivo culture experiments, ovaries were collected from WT and caspase-3 null female littermates at 2, 4, or 6 d post ovulation, and the occurrence of apoptosis within the CL was analyzed. Whereas ovaries of WT mice had only residual luteal tissue at d 6 post ovulation, ovaries collected from caspase-3-deficient mice retained many CL, at d 6 post ovulation, that were similar in size to those observed in the early luteal phase of WT mice. Importantly, there was no dramatic increase in apoptosis in CL of caspase-3-deficient mice at any time point examined post ovulation, indicating that the involution process had indeed been delayed. In contrast, the levels of progesterone declined regardless of genotype. These data provide the first direct evidence that caspase-3 is functionally required for apoptosis to proceed normally during luteal regression. However, caspase-3 is not a direct mediator of the decrease in steroidogenesis associated with luteolysis.

Animals↗

PubMed2004

Extracellular H+ dynamics during oogenesis in Rhodnius prolixus ovarioles.

The spatiotemporal dynamics of transmembrane hydrogen ion (H+) fluxes during oogenesis were investigated in the telotrophic ovarioles of the insect, Rhodnius prolixus. Although Rhodnius ovarioles possess exaggerated morphological and electrical polarity between nurse cells and oocytes, little is known about H+ changes during oogenesis, despite the regulatory role played by H+ and pH in many relevant cellular processes. A number of dynamic extracellular H+ fluxes were measured along Rhodnius ovarioles, representing an oogenesis cycle, using a non-invasive, self-referencing, H+-selective probe. The interfollicular stalk separating adjacent follicles exhibited prominent H+ efflux that peaked during midvitellogenesis and declined during late vitellogenesis. H+ efflux in this region preceded stalk formation and, importantly, preceded the onset of vitellogenesis in the adjacent posterior follicle. H+ efflux was also observed over the terminal follicle, where specialized regions of the chorion were forming, and was still detected around follicle cells after ovulation, indicating that the somatic follicular epithelium produced this flux. Transmembrane H+ fluxes may drive intracellular pH changes or may stabilize pHi in response to pH-altering events. H+ fluxes may play a role in processes that coincide spatially and temporally, including the onset of vitellogenesis, endocytosis, follicle cell cytoskeletal dynamics, and regulation of interfollicular feedback mechanisms.

Analysis of Variance↗

PubMed1983

Follicular growth and maturation in the domestic hen (Gallus domesticus).

The rate of follicular development in hens was assessed by measuring the increase in the diameter of follicles during 120 h before ovulation and the mass of follicles during 48 h before ovulation. The rate of follicular maturation was estimated by the ovulatory response of follicles to an injection of GnRH and the ability of follicles to produce progesterone in response to an injection of LH. Follicular diameter and follicular mass increased until ovulation, indicating that the follicle continues to sequester yolk material until ovulation occurs. The ovulatory response to GnRH and the production of progesterone in response to LH were negligible until 10 h after ovulation and then both aspects of follicular competence began to be functional. It was concluded that the size of the hen's ovum is not a primary factor regulating ovulability; rather, the acquisition of the ability to ovulate spontaneously is associated with the ability to produce progesterone in response to an LH stimulus. The ability to ovulate in response to GnRH was acquired more quickly in hens laying long sequences than in hens laying short sequences, indicating that the ovulatory response of the follicle can be used to assess the reproductive potential of laying hens.

Animals↗

PubMed1984

Pre-ovulatory arrest and peri-ovulatory redistribution of competent spermatozoa in the isthmus of the pig oviduct.

Using the surgical approach of post-coital ligation and transection of the distal oviduct at different times relative to ovulation, together with subsequent recovery of the eggs, gilts mated at the onset of oestrus were studied for progression of viable spermatozoa within the isthmus. Results are derived from 76 animals and examination of 1047 eggs. Transection of the isthmus 1.5-2.0 cm proximal to the utero-tubal junction at intervals from 3 to 36 h after mating prevented fertilization in 269 of 270 eggs, whereas 98% of 223 eggs were fertilized in the control oviducts. Transection at 38 h (pre-ovulatory), 40 h (peri-ovulatory) and 42-44 h (post-ovulatory) after mating yielded, respectively, 5%, 40% and 100% fertilization. The mean number of spermatozoa associated with the zona pellucida increased in a parallel manner. These results, and those obtained with ligatures placed closer to the site of fertilization just after ovulation, indicate a pre-ovulatory arrest of viable spermatozoa in the caudal region of the isthmus for 36 h or more followed by an active ad-ovarian redistribution.

Animals↗

PubMed2001

Relationship between maternal endocrine environment, early embryo development and inhibition of the luteolytic mechanism in cows.

In this study, the relationship between maternal hormone environment and early embryo development in mature non-lactating Holstein-Friesian cows was investigated. Animals were inseminated at either 72 or 96 h after prostaglandin injection (n = 23) or were left as uninseminated controls (n = 10). Plasma samples were collected once a day from the first day of insemination (day 1) until day 16, when the cows underwent an oxytocin challenge, and were then slaughtered and their reproductive tracts removed. The tracts were flushed to collect embryos and the flushes were measured for interferon tau (IFN-tau) activity. Inseminated cows without an embryo on day 16 (n = 5) underwent both delayed ovulation (indicated by delayed decrease in oestradiol concentrations) and a delayed increase in progesterone concentrations after ovulation compared with cows with an embryo on day 16 (n = 15). Within the group of cows with an embryo, those with poorly developed embryos producing undetectable concentrations of IFN-tau (n = 7) had similar oestradiol profiles but underwent a delayed progesterone increase after ovulation compared with cows with well developed embryos producing measurable quantities of IFN-tau (n = 8). In the cows with an embryo, the plasma concentration of 13,14-dihydro-15-keto PGF2a, the principal metabolite of PGF2a, after injection of oxytocin was lower than that of control cows and cows without an embryo. However, when the cows with an embryo were compared on the basis of production of embryonic IFN-tau, the PGF2a response to oxytocin was attenuated completely in cows that had measurable IFN-tau activity, whereas a response of similar magnitude to that in control cows and cows without an embryo was observed in those with undetectable IFN-tau activities. In conclusion, the successful maternal recognition of pregnancy in cows depends on the presence of a sufficiently well developed embryo producing sufficient quantities of IFN-tau, which is, in turn, dependent on an appropriate pattern of maternal progesterone secretion.

Animals↗

PubMed2000

Role of ovarian failure in reproductive senescence in aged red deer (Cervus elaphus) hinds.

Physiological and endocrine factors associated with reproductive senescence were assessed in a group of 19 ageing red deer hinds. Reproductive success, defined as the percentage of hinds weaning a calf successfully, decreased gradually from 89% at 6-7 years of age to 50% at 17 years, and subsequently decreased markedly; only one hind reared a calf at 19-20 years of age. When the 12 surviving hinds were approaching 21 years of age, they were compared with ten mature 7-year-old females over the onset of the breeding season. All hinds were subsequently killed, the reproductive tracts were recovered and antral (>/= 2 mm in diameter) and preantral follicle populations were determined by dissection (n = 7 hinds per age group) or stereological analysis (n = 2 ovaries per age group), respectively. Cyclical ovarian activity (plasma progesterone) was evident in fewer aged hinds compared with mature hinds (3/12 versus 10/10, P < 0.001) and mean plasma LH concentrations were higher in aged animals than in mature animals (0.57 +/- 0.05 and 0.20 +/- 0.05 ng ml(-1), P < 0.001). Mean uterine (44.2 +/- 4.5 and 75.4 +/- 4.2 g; P < 0.001) and ovarian masses (0.88 +/- 0.11 and 1.52 +/- 0.12 g; P < 0.001) were lower in the aged hinds, which also had fewer antral follicles than did mature hinds (0.89 +/- 0.35 and 23.5 +/- 4.5 follicles per hind, respectively; P < 0.001). Only one primordial follicle was observed in one of the ovaries of the aged hinds, compared with 7000-21 000 in the ovaries of mature hinds. The high gonadotrophin concentrations, paucity of primordial and antral follicles and failure of ovulation indicate collectively that waning reproductive performance after 17 years of age is primarily due to ovarian failure.

Aging↗

PubMed2003

Dynamic evaluation of ovarian reserve and abnormal androgen excess in women.

The evaluation of the ovarian reserve is an important area of clinical investigation that gives information on endowment and functional activity of remaining follicles within the ovary, thus concerning the female reproductive potential. In neonatal and pediatric age, an ovarian failure is commonly due to disorders of sexual differentiation. In adults, the basal and dynamic tests that predict the ovarian reserve are particularly useful in women undergoing assisted reproductive programs. Transvaginal ultrasound study of ovarian folliculogenesis performed simultaneously with the evaluation of cervical score, FSH, LH and estradiol plasma levels, evidentiates follicular rupture and ovulation, indicating also the optimum timing of hCG administration. Basal day 3 FSH, 17-beta-estradiol and inhibin B plasma levels give information on the ovarian potential. Clomiphene citrate challenge test (CCCT) and GnRH agonist stimulation test (GAST) have clinical utility as indicators of ovarian reserve but their accuracy does not allow to be predictive in terms of number-per-unit tissue of the remaining follicle within the ovary. In the present paper the strategies to study hyperandrogenism and polycystic ovarian syndrome, a common cause of ovarian reserve reduction and subfertility, are also reviewed. The abnormal androgen excess in women can be referred to the ovary, the adrenal glands, or the peripheral conversion of androgen precursors. Dynamic tests may be useful for determining the amount of androgens rising from each of these sites helping the therapeutic strategies.

Adolescent↗

PubMed1991

A comparative study on involvement of tPA activity in ovulation induced by hCG and GnRH agonist in hypophysectomized rats.

A GnRH agonist (5-50 micrograms) is capable of inducing ovulation in PMSG-primed hypophysectomized immature rats, as is the case in hCG-induced ovulation, but 2-4 h earlier than hCG. GnRH-induced ovulation is effectively blocked by the concomitant administration of the GnRH-antagonist which failed to interfere with hCG-induced ovulation, indicating that GnRH and its agonists do not share a receptor with LH/hCG. Like hCG, GnRH is also capable of inducing tissue type (tPA), but not urokinase type (uPA) PA. The plasminogen activator activity in ovarian homogenates and the granulosa and theca-interstitial cells increase in a time-dependent manner, reaching maximum levels just prior to ovulation. Similar to hCG, GnRH also increases tPA activity in cumulus-oocyte complexes in a time-dependent fashion.

Animals↗

PubMed1979

Precoital single doses of delta9-tetrahydrocannabinol block ovulation in the rabbit.

delta9-Tetrahydrocannabinol (delta9-THC) inhibits pituitary gonadotropin secretion in castrated rhesus monkeys. Also, delta9-THC blocks the ovulatory reflux in rabbits. We report now the dose-response relationship of precoital single doses of delta9-THC on luteinizing hormone (LH) release and ovulation in the rabbit. Forty-five female rabbits in estrous were divided into nine groups of five animals. Groups 1 to 5 received a single intramuscular dose of delta9-THC (5,2.5, 1.25, 0.612, and 0.306 mg/kg, respectively) 2 hours before mating; animals of group 6 received vehicle only. In animals of groups 7 and 8 ovulation was induced with 100 IU of human chorionic gonadotropic (hCG), given intravenously 2 hours after the administration of delta9-THC (5 mg/kg) or vehicle. Rabbit luteinizing hormone (rLH) was measured in plasma 90 to 120 minutes after coitus or hCG administration. After the injection of 5 mg of delta9-THC, luteinizing hormone-releasing factor (LH-RF) (20 microgram intravenously) was administered to the animals of group 9. All animals of groups 6, 7, and 8 ovulated. A dose-response curve was observed in the animals treated with delta9-THC and natural mating. Whereas none of the animals treated with 5 or 2.5 mg/kg ovulated, one of the group treated with 1.25 mg/kg, two of the group treated with 0.612 mg/kg, and all treated with 0.312 mg/kg ovulated. Ovulations correlated with postcoital levels of rLH. All animals of group 9 ovulated, indicating that the site of action of delta9-THC is suprapituitary, probably hypothalamic.

Animals↗

PubMed1997

Effect of alcohol on the proestrous surge of luteinizing hormone (LH) and the activation of LH-releasing hormone (LHRH) neurons in the female rat.

Reproduction is adversely affected by alcohol abuse in humans and laboratory animals. In rats, alcohol exposure suppresses both luteinizing hormone (LH) and sex steroid secretion, although consensus is lacking as to which level of the hypothalamic-pituitary-gonadal (HPG) axis is primarily affected. We tested the hypothesis that acute alcohol treatment inhibits the HPG axis by blunting release of LH-releasing hormone (LHRH) in female rats, by examining the effect of this drug on the central reproductive endocrine event; i.e., the proestrous surge of gonadotropins, which triggers ovulation. In a first series of experiments, we injected alcohol at 8 A.M. and 12 P.M. on proestrus and measured plasma levels of LH, estradiol (E2), and progesterone during the afternoons of proestrus and estrus. Alcohol administration blocked the proestrous surge of LH and ovulation. In subsequent experiments, alcohol inhibited the surge of LHRH (measured by push-pull cannulation) and LHRH neuronal activation (measured by Fos labeling in LHRH neurons). Because alcohol also decreased E2 levels, we reasoned that it might have prevented positive feedback; however, alcohol retained its ability to inhibit the LH surge evoked by E2 implantation in ovariectomized females, disproving this hypothesis. Additionally, alcohol does not act via increased corticosteroid secretion, because alcohol also blocked the proestrous surge in adrenalectomized females. Last, exogenous administration of LHRH to alcohol-blocked animals evoked LH secretion and ovulation, indicating that pituitary and/or ovarian function could be restored by mimicking the hypothalamic signal. Collectively, these data indicate that in female rats, alcohol inhibits the gonadotropin surge primarily by decreasing LHRH secretion.

Adrenalectomy↗

PubMed1997

A study of early pregnancy factor activity in preimplantation.

PROBLEM AND METHOD: Early pregnancy factor (EPF), an Immunosuppressive substance, which appears in pregnant women's sera 48 h after fertilization, is a kind of pregnancy-specific protein. To determine whether the EPF activity could be a super early indicator of pregnancy, we used rosette inhibition assay to detect EPF activity in the sera, collected from 70 women 2-7 days after ovulation intending to conceive monitored by ultrasonography. Simultaneously we selected 40 non-pregnant sera and 12 early-pregnant sera as negative control and positive control, respectively. RESULTS: The results of this study demonstrated that EPF activity is detected in 35 women's sera out of 70 women within 2-7 days after ovulation, and 28 women out of the 35 were pregnant, which was known by follow-up, and 7 were not pregnant, possibly due to either false positive results or embryo loss because of preimplantation failure, thus causing no pregnancy. The other 35 out of 70 had no EPF activity and 34 of them were not pregnant, which was known by follow-up, but one case became pregnant, which was false negative result. Our study showed that diagnosis of the super early pregnancy could be made by detecting EPF activity in maternal serum within the time of preimplantation. The accuracy of pregnancy diagnosis by this method is 88.6%, with a false negative rate of 3.4% and a false positive rate of 17.1%. The beta-HCG level was measured from the above 70 women's sera in order to contrast EPF activity. All of the sera collected 2-6 days following ovulation indicated that there were lower beta-HCG values in very early pregnancy (> or = a5 mIU/ml). On the seventh day after ovulation, EPF activity was detected in 11 out of 15 sera with only 2 of them with a b-HCG level that reached or slightly surpassed that of the early pregnancy diagnosis (5 mIU/ml and 5.4 mIU/ml, respectively). This demonstrated that beta-HCG is not the earliest signal of pregnancy; otherwise the EPF activity is one that appears 2-6 days earlier than beta-HCG appears. We measured the progesterone level of the 48 sera from the 70 collected above within 2-7 days postovulation and found most of them reached the level of progesterone in the luteal phase (7.5-98.3 nmol/L). This indicated that ovulation had taken place in these women, which was in accordance with observations by ultrasonography. CONCLUSIONS: Our study showed that diagnosis (of 88.6%) of super early pregnancy could be made with an accuracy of 88.6% by detecting EPF activity in maternal serum within 2-days after ovulation. This offers a basis for pregnancy diagnosis for the women who attempt to terminate their pregnancy safely or who conceive unexpectedly, and it contributes to family-planning.

Adult↗