Early Pregnancy Development Research Articles

Macrophages Are Essential for Maintenance of Corpus Luteum Function in Early Pregnancy.

Macrophages are abundant within the ovary, and have been identified in the corpus luteum (CL) of most species studied, including in rodents and human. Through their secretory products, macrophages are thought to be involved in ovarian tissue remodelling, including luteinization, and in regulating steroidogenesis. Macrophages co-cultured with granulosa or luteal cells act to stimulate progesterone secretion in vitro. To determine the impact of macrophage ablation during early pregnancy, we utilised CD11b-DTR transgenic mice to elicit transient systemic ablation of macrophages by administration of diphtheria toxin (DT). The effects of macrophage ablation during the pre-implantation phase of pregnancy were evaluated. Ablation of macrophages on day 1 pc or day 4 pc caused complete pregnancy loss in all DT-treated CD11b-DTR mice, while DT-treated wild-type mice or PBS-treated CD11b-DTR mice maintained viable pregnancies. Macrophage ablation on day 3 pc significantly reduced serum progesterone (P4) levels to 40% of control levels when measured 24 h later. Administration of exogenous P4 on each of day 4-7 pc prevented fetal loss in DT-treated CD11b-DTR mice, and pregnancy progressed with viable pups at late gestation, while no pregnancies remained viable in DT-treated mice administered vehicle only. Immunohistochemical evaluation with anti-CD31 mAb showed that macrophage ablation in the CD11b-DTR mice resulted in disappearance of endothelial cells from within the CL. This suggests that macrophages are essential for support of endothelial cells within the CL, either by provision of trophic support and/or by macrophage transdifferentiation into endothelial cells. In conclusion, these data indicate a critical role for macrophages in corpus luteum development and steroidogenic function in early pregnancy. Given that various nutritional and immune stressors can profoundly affect macrophage numbers and behaviour, it seems reasonable to speculate that macrophage-regulated CL development is a vulnerable event and may contribute to some forms of unexplained infertility. (platform)

Endometrial implantation factors in women with submucous uterine fibroids

Uterine fibroids are benign tumours, which are associated with subfertility and early pregnancy loss. This study was carried out to examine the effect of submucous fibroids on concentrations of glycodelin, insulin-like growth factor binding protein-1 (IGFBP-1), interleukin-6 (IL-6), interleukin-10 (IL-10), tumour necrosis factor α (TNFα) and osteopontin in uterine flushings. Premenopausal women with a certain diagnosis of submucous fibroid confirmed on three-dimensional saline infusion sonohysterography were recruited into the study. The control group included women without ultrasonic evidence of any uterine or endometrial pathology. All women had uterine flushings performed 7days post LH surge. Enzyme linked immunoassays were performed to analyse glycodelin, IL-6, IL-10, TNFα and osteopontin, whilst immunoradiometric assay was used to analyse IGFBP-1. In 23 women with submucous fibroids, the concentrations of glycodelin and IL-10 in uterine flushings were significantly lower compared with 17 women in the control group (P=0.002; P=0.007, respectively). There were no significant differences between the two groups in concentrations of IGFBP-1, IL-6, TNFα and osteopontin. Women with submucous fibroids had significantly lower concentrations of glycodelin and IL-10 in mid-luteal phase uterine flushings. This finding may explain the association with submucous fibroids and adverse reproductive outcomes. Uterine fibroids are small growths from the muscle of the uterus (womb). Submucous fibroids protrude into the cavity of the womb. We do not know what causes fibroids to form and grow. In most women, fibroids cause no symptoms and they are sometimes detected on routine gynaecological examination. In some women, however, fibroids can cause heavier and longer menstrual periods. Another problem associated with fibroids is bleeding between periods. The effect of fibroids on fertility is not clear, but some doctors believe that they may also cause infertility and early miscarriage. This study tried to see whether presence of submucous fibroids has any effect on various substances produced by the lining of the womb to facilitate development of early pregnancy. Women with a confirmed diagnosis of submucous fibroids were asked to attend the clinic and have the uterine cavity flushed with a special solution 7days after ovulation. The fluid, which was taken back from the womb, was then analysed to measure the amounts of substances that favour pregnancy development. Women with a normal uterine cavity were also asked to have the uterine cavity flushed to act as a comparison. The study showed that the uterine cavities of women with submucous fibroids were producing decreasing amount of substances favourable to early pregnancy development. We speculate that this may explain why some women with submucous fibroids have difficulties falling pregnant. Our findings should be helpful to doctors advising women with submucous fibroids who wish to start a family.

Secretion of Angiogenic Growth Factors by Villous Cytotrophoblast and Extravillous Trophoblast in Early Human Pregnancy

Angiogenesis is a key feature of placental and uterine development in early pregnancy. We hypothesized that trophoblast cells produce angiogenic growth factors, and that expression differs between villous cytotrophoblast (CTB) and extravillous trophoblast (EVT) cells. Fourteen angiogenic growth factors were measured by multiplex growth factor analysis or ELISA in tissue culture supernatants from EVT and CTB from pregnancies at 8–10 and 12–14 weeks' gestation. Gestational age and cell type differences were observed. EVT and CTB are major producers of angiogenic growth factors that likely contribute to placental vascular development and spiral artery remodeling.

Ultrasound Evaluation of Pregnancy in the First Trimester

Abstract Improvements in ultrasound technology, including transvaginal sonography and higher frequency probes, have led to a better understanding of early pregnancy development. These advances and the increasing availability of ultrasound allow women to have an earlier and more accurate assessment of their pregnancy. First trimester sonographic signs have been identified that can be used to reassure women that their pregnancy is progressing normally or counsel them that their pregnancy will fail. In addition, first trimester ultrasound can accurately predict the type of twinning in multiple gestations, allowing for appropriate counseling and management. Objectives Know the first trimester ultrasound findings of a normal intrauterine pregnancy Understand the ultrasound findings that diagnose an early pregnancy failure Know the ultrasound criteria used to diagnose a multifetal gestation

The development of children and the health of societies

The development of children and the health of societies

Assessment and Management of Bleeding in the First Trimester of Pregnancy

Vaginal bleeding occurs in 15% to 25% of early pregnancies. While 50% of women who have vaginal bleeding in the first trimester of pregnancy will continue to have a viable pregnancy, the event creates significant anxiety for the woman and can be managed in a multitude of ways. The 3 main differential diagnoses associated with vaginal bleeding are spontaneous abortion, ectopic pregnancy, and gestational trophoblastic disease. This article reviews early pregnancy development, etiologies of vaginal bleeding in the first trimester, strategies for evaluation, and recognition and management of the main diagnostic considerations. Case study examples illustrating the complexity of the assessment and management of vaginal bleeding in early pregnancy are presented.

“Extra Amniotic Band Syndrome” – Uterine Synechiae and Development of Early Pregnancy

Study Objective: Curettage, uterine surgery or puerperal interventions, can cause synechiae. Synechiae vary in consistency (flimsy, intermediate or strong), position (cervical, fundal) and area engaged. Synechiae can disappear or remain during pregnancy. Flimsy and intermediate adhesions will stretch/break, whereas strong synechiae force the amniotic sac around it, creating an amniotic fold. Entrapment of the fetus can be seen by ultrasound. Severe deformation of the amniotic sac can influence fetal development. Those deformations show resemblance with abnormalities as seen in amniotic band syndrome No theories concerning amniotic band syndrome (ABS) can explain all features. We hypothesise that in case of development of the fetus between strong, multiple and conglomerated synechiae, occurring fetal deformities can mimic features as seen in ABS. Arguments and animations are demonstrated to support this hypothesis. Study Objective: Curettage, uterine surgery or puerperal interventions, can cause synechiae. Synechiae vary in consistency (flimsy, intermediate or strong), position (cervical, fundal) and area engaged. Synechiae can disappear or remain during pregnancy. Flimsy and intermediate adhesions will stretch/break, whereas strong synechiae force the amniotic sac around it, creating an amniotic fold. Entrapment of the fetus can be seen by ultrasound. Severe deformation of the amniotic sac can influence fetal development. Those deformations show resemblance with abnormalities as seen in amniotic band syndrome No theories concerning amniotic band syndrome (ABS) can explain all features. We hypothesise that in case of development of the fetus between strong, multiple and conglomerated synechiae, occurring fetal deformities can mimic features as seen in ABS. Arguments and animations are demonstrated to support this hypothesis.

Can Early hCG Levels be a Marker for Pregnancy Outcome in ART Cycles?

ABSTRACT With advancing technology of assisted reproduction, physicians today have the ability to achieve conception in many couples who would have been totally incapable doing so only a few years ago. The anxiety and the uncertainty of pregnancy outcomes using ART procedures is widely accepted as one of the main psychological stresses the couples. The ability to predict outcome as soon as possible after assisted conception treatment is important for clinic staff and patients. The aim of this observational study is to highlight the importance of hCG values in predicting the outcome of ART cycle and counseling the patients in case of adverse result. The ultimate aim is to improve the take home baby rate and initial hCG value can help us counsel our patients towards the ultimate outcome. Embryo development in early pregnancy follows a preprogrammed-timing schedule and depends mainly on the embryonic age of the healthy, successfully implanted conceptus. The appearance of hCG in maternal serum is used to assess the time of clinically detectable implantation. bhCG has provided the best sensitivity and specificity for detection of normal and pathological pregnancies. After IVF, early pregnancy loss or multiple gestations may be predicted with high sensitivity and specificity by using cut-off values of serum hCG. The median HCG concentration was 116 IU/l in viable pregnancies and 31 IU/l in nonviable pregnancies. The median hCG concentration in twin pregnancies was almost double that in singleton pregnancies (201 IU/l vs 116 IU/l). Thus we can reassure normally pregnant patients as well as filter and manage those with nonviable outcomes more efficiently.

Ultrasound Evaluation of Abnormal Early Pregnancy

Abstract Ultrasound examination has become the “golden standard” in follow-up of the development and complications in early pregnancy. With introduction of transvaginal sonography a possibility for early morphological and biometrical ultrasound examinations has been significantly improved. The essential aim of an early pregnancy ultrasound is not only to diagnose a pregnancy, but also to differentiate between normal and abnormal pregnancy. Application of color Doppler ultrasound has enabled functional hemodynamic presentation and evaluation soon after implantation.

Concentration of chemokines in peripheral blood in first trimester of diabetic pregnancy

Several types of regulators (i.e. chemokines and metalloproteinases) are considered to play a crucial role in pregnancy by local modulation of the immune system at the level of peripheral leukocytes. The aim of this study was to determine whether changes in chemokines (interferon-gamma-inducible protein (IP-10), monocyte chemotactic peptide-1(MCP-1), cytokines regulated upon activation normal T cell expressed and secreted (RANTES) and matrix metalloproteinase-9 (MMP-9)) concentrations in diabetic patients could affect the course of pregnancy. The study group consisted of 65 diabetics in the first trimester of pregnancy. Some 47 pregnancies were successfully continued to delivery, 18 were terminated with spontaneous miscarriages. Twenty healthy women matched for gestational age served as a control group. Glycated haemoglobin (HbA1C), vascular complications and lipoproteins (cholesterol, HDL-cholesterol, low density lipoprotein (LDL)-cholesterol and triglicerides) concentrations in maternal blood did not influence the chemokines concentrations. Lower RANTES level and higher MMP-9 concentrations were found in diabetic women. MCP-1 and RANTES levels differed significantly between pregnancies with good and poor perinatal outcome. A logistic regression model revealed that not only duration of diabetes, age of patients, HbA1C and insulin requirements, but also MMP-9,RANTES, MCP-1 and LDL-cholesterol levels seem to be involved in first trimester metabolism. Our results suggest the possible role of chemokines in early pregnancy development, especially in well-controlled diabetic patients, when hyperglycaemia is unlikely to be the main reason for an unfavourable outcome. Our results show that MCP-1 and RANTES might serve as predictive factors for an unfavourable outcome in diabetic pregnancy, whereas MMP-9 seems to be a marker of immunological changes related to mild hyperglycaemia. However, the open question of how the modulation of chemokines concentrations might be applied to prevent miscarriage in diabetic patients remains.

High BMI is an independent predictor of unsuccessful fresh blastocyst transfer

To determine the predictive value of patient and cycle characteristics for clinical pregnancy following blastocyst transfer. Retrospective study 2003–2006. Table 1Summary of Patient Characteristics by Clinical OutcomeCharacteristicsClinical PregnancyNot PregnantP ValueAge32.6 ± 3.634.0 ± 3.90.10BMI (kg/m2)24.3 ± 4.426.1 ± 4.60.09Max FSH (mIU/mL)6.9 ± 2.16.6 ± 1.90.54# Blastocysts Transferred2.1 ± 0.52.3 ± 0.70.24Patients in the highest tertile for BMI were less likely to achieve CP following blastocyst ET (OR 2.7, 95% CI 0.9 - 9.5), Figure 1. Multivariate logistic regression analysis adjusting for age and number of blastocysts transferred confirmed low BMI as an independent negative predictor of CP. Open table in a new tab Patients in the highest tertile for BMI were less likely to achieve CP following blastocyst ET (OR 2.7, 95% CI 0.9 - 9.5), Figure 1. Multivariate logistic regression analysis adjusting for age and number of blastocysts transferred confirmed low BMI as an independent negative predictor of CP. ART cycles proceeding to fresh blastocyst embryo transfer (ET) were included (n = 75). As per our protocol, decision to proceed to blastocyst ET was based on patient's age and embryo cleavage status. Donor oocyte derived blastocyst ET were excluded. Outcome of interest was clinical pregnancy (CP); negative CP category incorporated non-pregnant, biochemical, and ectopic pregnancies. Associations of CP with patient characteristics (age, BMI, maximal FSH) and cycle parameters (peak E2, # eggs, available cleaving embryos on Day 2 and Day 3, # blastocysts, # blastocysts ET, and # cryopreserved) were evaluated using Student's T test, Mann Whitney rank sum test, and Pearson Correlation. BMI was categorized in tertiles for evaluating an association with CP. Multivariate logistic regression models were created to determine independent predictors of CP following blastocyst ET. P value <0.05 was considered statistically significant. Other than an inverse association of borderline significance noted between BMI and CP, additional patient and cycle parameters were comparable between groups by pregnancy outcome (Table 1). Mean BMI (mean ± SD): tertile 1 (20.9 ± 1.5 kg/m2), tertile 2 (24.4 ± 1.1 kg/m2), and tertile 3 (30.1 ± 4.0 kg/m2). The data suggest that a high BMI exerts a detrimental influence on implantation or early pregnancy development, furthering evidence that obesity is associated with subfertilty.

VEGF receptor-2 (VEGFR-2) and VEGFR-3 mediate peri-implantation uterine angiogenesis during early pregnancy development

Induction of angiogenesis in the uterine decidua is a key event in peri-implantation uterine differentiation and blastocyst implantation. VEGFR-2 is expressed on vessels in the periphery (secondary decidual zone), and to a lesser extent, in the area surrounding the embryo (primary decidual zone). In contrast, VEGFR-3 expression is restricted to vessels in the primary decidual zone. Given the distinct expression patterns, we hypothesized that VEGFR-2 and VEGFR-3 make unique contributions to peri-implantation uterine angiogenesis.

P-551: Blockage of the VEGF-C/VEGF receptor 3 pathway disrupts early pregnancy development in the rodent

A major regulator of angiogenesis and/or lymphangiogenesis is VEGF-C acting through the VEGF receptor 3 (VEGFR-3). It is currently not known whether functionality of this pathway is important in the peri-implantation period. We tested whether a single dose of a blocking antibody against VEGFR-3 (mF4-31C1) on embryonic day (ED) 3.75 might interfere with normal embryonic development. Prospective animal laboratory. Adult pregnant female CD1 mice were randomized into 5 groups: baseline: sacrificed on ED 3.75; control: saline injection on ED 3.75; replacement control: Subcutaneous placement of three 10 mg/pellet 21-day release progesterone (P4) tabs (Innovative Research of America) on ED 3.5, and saline injection on ED 3.75; treatment: anti-VEGF-R3 antibody mF4-31C1 injection (160 mg/kg; ImClone Systems, New York) on ED 3.75; replacement plus treatment: Three P4 tabs ED 3.5, and anti-VEGF-R3 antibody ED 3.75. Animals were sacrificed on ED 6.5 or 13.5 (n=5 per group). Outcome parameters included: uterine and ovarian histology immunofluorescence (antibodies against the endothelial cell marker PECAM and against VEGFR-3), uterine weights, number of implantation sites, embryonic development, and maternal P4 levels. The Wilcoxon rank test was used for statistical analysis. PECAM staining mostly colocalizes with staining for VEGFR-3 when analyzed on ED 6.5 indicating that this receptor is primarily found on small vasculature in ovarian corpora lutea and uterus. Uterine weights increased in control and replacement control animals from 0.27±0.02g (ED 3.5) to 6.4±0.9g (ED 13.5) with the progression of pregnancy, whereas no weight increase was observed in the treatment and replacement plus treatment groups on ED 13.5 (p<0.01). Implantation sites were mostly absent on ED 13.5 in the treatment and the replacement plus treatment group. Histological evaluation of the uterus revealed the presence of minimal necrotic tissue with or without old blood on ED 13.5. In contrast in control and replacement control animals appropriate embryonic and placental structures were observed on ED 13.5. Endothelial cell staining with PECAM demonstrated that luteal vascular density (ovary) was similar in all groups on ED 6.5. In contrast uterine blood vessel density evaluated on ED 6.5 decreased in the two treatment groups by 25% when compared to controls (p<0.05). Progesterone (ED 13.5) levels fluctuated around 25 ng/ml in all groups on ED 13.5. Anti-VEGFR-3 antibody administered in the peri-implantation period interfered with in-vivo pregnancy development. The antibody disrupted formation and maintenance of peri-implantation blood vessels, possibly decreasing the blood supply to the developing embryo. In contrast the antibody did not affect ovarian P4 secretion or luteal endothelial cells. Interestingly, P4 replacement did not reverse the effects of the antibody, and disruption of pregnancy development occurred. Therefore, the VEGF-C/VEGFR-3 pathway seems to be important for early pregnancy development in the rodent. One mechanism of action seems to be the regulation of angiogenesis in the decidua during the peri-implantation period.

Factors affecting the early embryonic environment

The early human embryo develops in a tightly controlled, relatively protected environment. During the first 8–12 weeks of human gestation, the delivery of maternal blood, and therefore the amount of oxygen to the developing embryo is limited and the embryo is supplied with essential nutrients in part via the decidual uterine glands, the placental trophoblast and the secondary yolk sac. Factors that interfere with this process may well result in spontaneous miscarriage or adverse outcome later in pregnancy. There is mounting evidence for the presence of transporter systems for many substances including drugs and toxins from the maternal to the fetal compartments in early pregnancy. The role of these substances in both the protection of early pregnancy development and possible teratogenicity are explored in this chapter. Clearly, the timing of exposure to potentially damaging substances is crucial with effects on conception, implantation, placental development and organogenesis dependent on the gestation at which exposure occurs.

Maternal Serum C-Reactive Protein Concentration Early in Pregnancy and Subsequent Pregnancy Loss

The purpose of this study was to determine the relationship between maternal inflammation and first or second trimester pregnancy loss. A nested case-control analysis was performed among the cohort enrolled in the Oral Conditions and Pregnancy study. We compared maternal serum C-reactive protein concentration between women with a pregnancy loss at < 21 weeks gestation to control women without gestational diabetes or preeclampsia who delivered at term. Participants were 2:1 frequency matched to cases by maternal age, race, and prior preterm birth. Median concentration of serum C-reactive protein between cases and controls was compared using Wilcoxon rank sum test. The potential effects of maternal smoking, gestational age at blood draw, and insurance status were evaluated and an adjusted odds ratio (95% confidence interval) for pregnancy loss was calculated using multivariable logistic regression. Among 1224 participants, 102 (9.8%) experienced pregnancy loss and 44 had complete information available. Median serum C-reactive protein concentration was significantly higher in controls compared with all cases (3.2 versus 0.5 microg/mL; p < 0.001). However, when stratified by gestational age at the time of the loss, median serum C-reactive protein level among controls was similar to those with a loss at less than 12 weeks (3.2 versus 2.0 microg/mL) but significantly higher compared with those whose loss occurred at greater than 12 weeks gestation (3.2 versus 0.5 microg/mL; p < 0.05). After adjusting for maternal smoking, gestational age at blood draw, and insurance status, women whose serum C-reactive protein level was greater than the 75% percentile had a decreased odds ratio for pregnancy loss (0.20; 95% confidence interval, 0.06 to 0.65). Pregnancy loss is not associated with increased systemic inflammation as measured by maternal serum C-reactive protein. Future study should be directed at determining the role of maternal inflammation during early pregnancy development and placentation.

The role of ultrasound imaging in diagnosing and investigating early pregnancy failure

The advent of high-resolution transvaginal ultrasound (TVS) has revolutionized our understanding of the pathophysiology and the management of early pregnancy failure. Knowledge of the ultrasound appearances of normal early pregnancy development and a good understanding of its pitfalls are essential for the diagnosis and management of early pregnancy failure. Ultrasound imaging has rapidly replaced all other techniques used to study normal human development in the first trimester, and ultrasound features of the early gestational sac have corroborated anatomical studies showing that the first structures to appear are the celomic cavity and the secondary yolk sac. No single ultrasound measurement of the different anatomical features in the first trimester has been shown to have a high predictive value for determining early pregnancy outcome. Similarly, Doppler studies have failed to demonstrate abnormal blood flow indices in the first-trimester uteroplacental circulation of pregnancies that subsequently end in miscarriage. Ultrasound parameters combined with maternal serum hormone levels, maternal age, smoking habits, obstetric history and the occurrence of vaginal bleeding have all been combined in multivariate analyses, with mixed results. Combined ultrasound and in-vitro experiments have demonstrated that the maternal circulation inside the placenta starts at the periphery at around 9 weeks of gestation and that this is associated with a physiological oxidative stress which could be the trigger for the formation of the placental membranes. Abnormal development of these membranes can result in subchorionic hemorrhage and threatened miscarriage with subsequent long-term consequences such as preterm rupture of the membranes and preterm labor, irrespective of the finding of a hematoma on ultrasound. In both euploid and aneuploid missed miscarriages there is clear ultrasound evidence for excessive entry of maternal blood at a very early stage inside the developing placenta resulting in oxidative stress and subsequent degeneration of villous tissue. The finding of blood flow in the intervillous space in cases of first-trimester miscarriage using color Doppler also appears to be useful in the prediction of success of expectant management. Miscarriages with blood flow within the intervillous space are up to four times more likely to complete with expectant management. TVS is considered the gold standard in the diagnosis and management of incomplete miscarriage. Expectant management of miscarriage, using ultrasound parameters to determine eligibility, could significantly reduce the number of unnecessary evacuations of the retained products of conception, depending on the criteria used.

Amino Thiols, Detoxification and Oxidative Stress in Pre-Eclampsia and Other Disorders of Pregnancy

New knowledge of placental development and function suggests that several common complications of pregnancy could share a similar origin. It is suggested that impaired placental development in early pregnancy may lead to placental oxidative stress and subsequently to the maternal syndromes such as recurrent early pregnancy loss and pre-eclampsia. Oxidative stress has been most extensively investigated in pre-eclampsia, resulting in hundreds of publications and many reviews. In general the literature points to the presence of placental and maternal oxidative stress. However, conformity amongst the relevant data is not absolute, most probably the result of the diversity of biomarkers investigated and the methods employed to assess oxidative stress, which generally depend on the assessment of end products of oxidative stress. Recently, new techniques have been developed that use different approaches based on the "real-time" measurement of oxidative stress by the redox status of thiols or the assessment of superoxide generation, whereas the role of Phase I/Phase II biotransformation pathways in oxidative stress was recognised. This review focuses on this biotransformation system, the thiol redox status and the involvement of these systems in oxidative stress associated with reproduction and pregnancy disorders, with the emphasis being laid on the syndrome of pre-eclampsia.

Biliary lithiasis in early pregnancy and abnormal development of facial and distal limb bones (Binder syndrome): A possible role for vitamin K deficiency

Binder syndrome is a maxillonasal dysostosis characterized by midface and nasal hypoplasia, sometimes associated with short terminal phalanges of fingers and toes and transient radiological features of chondrodysplasia punctata. Warfarin- or phenytoin-induced vitamin K deficiency during early pregnancy is a well-established etiology for this syndrome, which occurs nevertheless sporadically in most cases. We describe here the first case, to our knowledge, of Binder syndrome in a child whose mother presented with biliary lithiasis in early pregnancy. The mother proved to have a decrease in clotting factors II, VII, and X, and in prothrombin time, at 11 weeks of gestation, which was highly suggestive of vitamin K deficiency. The biliary lithiasis-induced vitamin K deficiency in early pregnancy is likely to have resulted in Binder syndrome. This observation should prompt physicians to carefully check for vitamin K deficiency in pregnant women presenting with biliary lithiasis, in order to prevent Binder syndrome in the fetus by providing intravenous vitamin K supplementation as soon as possible. Finally, reassuring genetic counseling regarding the genetic risk for future pregnancies is to be provided to the parents.

Regulation of Placental Vascular Endothelial Growth/Permeability Factor Expression and Angiogenesis by Estrogen during Early Baboon Pregnancy

We have recently shown that there was a developmental increase in placental trophoblast vascular endothelial growth/permeability factor (VEG/PF) expression and vascularization that closely paralleled maternal serum estrogen levels during advancing baboon gestation. The present study determined whether estrogen regulates these important aspects of primate development. VEG/PF mRNA levels were determined by competitive RT-PCR in isolated villous placental cells, and placental vascularization was assessed by image analysis. Placentas were obtained on d 60 of gestation (length of gestation is 184 d) from baboons in which estrogen levels on d 25-59 were increased by daily administration of aromatizable androstenedione or decreased by aromatase inhibitor CGS 20267. Androstenedione treatment increased maternal serum estradiol levels 3-fold (P < 0.01) and placental villous cytotrophoblast VEG/PF mRNA level to a value (mean +/- se, 26,836 +/- 5,625 attomoles/microg total RNA) 2.5-fold greater (P < 0.05) than that in untreated animals (11,645 +/- 1,746 attomoles/microg RNA). In contrast, administration of CGS 20267 decreased serum estradiol (P < 0.01) and placental cytotrophoblast mRNA (2,912 +/- 693 attomoles/microg RNA; P < 0.05) levels by 75%, effects prevented by concomitant administration of CGS 20267 and estradiol. VEG/PF mRNA levels in inner villous cells were unaltered. Coinciding with the increase in placental VEG/PF expression, the percent vascularized area (3.46 +/- 0.23) and vessel density (493 +/- 34 vessels/mm(2)) of the villous placenta in untreated baboons on d 60 were increased (P < 0.01) in baboons in which estrogen levels were elevated by androstenedione treatment (6.54 +/- 0.56 and 743 +/- 27 vessels/mm(2), respectively). It is concluded that estrogen has an important role in stimulating trophoblast VEG/PF expression and consequently villous placental angiogenesis to promote fetal growth and development in early primate pregnancy.

Effects of short-term early gestational exposure to endophyte-infected tall fescue diets on plasma 3,4-dihydroxyphenyl acetic acid and fetal development in mares.

Consumption of wild-type (toxic) endophyte-infected tall fescue (E+) by horses during late gestation is known to adversely affect pregnancy outcome; however, little is known of the potential disruptive consequences of E+ consumption by mares during the critical phases of placentation and fetal development in early pregnancy. The objective of this study was to evaluate the detrimental effects of feeding E+ to mares during early gestation. Mares (n = 12) paired by stage of gestation (d 65 to 100) were assigned to diets (six per diet) consisting of endophyte-free (E-) or E+ tall fescue seed (50% E- or E+ tall fescue seed, 45% sweet feed, and 10% molasses fed at 1.0% of BW/d). Mares also had ad libitum access to E+ or E- annual ryegrass hay, and were fed diets for 10 d. Following removal from the tall fescue diet on d 11, mares were placed on common bermudagrass pasture and monitored until d 21. Morning and evening rectal temperatures were recorded and daily blood samples were collected for progesterone and prolactin (PRL) analyses, whereas samples for 3,4-dihydroxyphenyl acetic acid (a catecholamine metabolite) analysis were collected on alternate days. For clinical chemistry analysis, blood samples were collected on d 0, 5, 10 and 21. Daily urine samples were collected for ergot alkaloid analysis, and ultrasonography was performed for presence of echogenic material in fetal fluids. Rectal temperatures (E+ 37.76+/-0.03; E- 37.84+/-0.03 degrees C) and serum PRL concentrations (E+ 14.06< or =0.76; E- 12.11+/-0.76 ng/mL) did not differ (P = 0.96) between treatments. Measuring the change in basal serum concentration from d 0 over time, progesterone concentrations did not differ (-0.64 +/-1.49 and -0.55+/-1.47 ng/mL for E+ and E- mares, respectively). There was no negative pregnancy outcome, and ultrasonography indicated no increase in echogenic material in fetal fluids. Plasma 3,4-dihydroxyphenyl acetic acid concentrations decreased (P < 0.05) in E+ compared with E- mares (2.1+/-0.14 and 4.4+/0.43 ng/mL, respectively). Urinary ergot alkaloid concentration was greater (P < 0.01) in mares consuming E+ compared with E- (532.12+/- 52.51 and 13.36+/-2.67 ng/mg of creatinine, respectively). Although no fetal loss was observed during the current study, elevated concentrations of urinary ergot alkaloid were consistent with depressed endogenous catecholamine activity, suggestive of an endocrine disruptive effect of hypothalamic origin.