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JE Eckert
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KL Gatford
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BG Luxford
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RG Campbell
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PC Owens
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Birth weight is a determinant of blood leptin concentrations in adults. Since nutrition during pregnancy can affect birth weight, the hypothesis that feed intake during pregnancy alters leptin expression in progeny was examined. Leptin mRNA was measured in subcutaneous adipose tissue and leptin protein was measuredin blood plasma from 59 day old female pigs whose mothers were fed at the same restricted rate except that half were permitted to consume 35% more feed during the second quarter of pregnancy. Leptin mRNA abundance in adipose tissue (P=0.015) and plasma leptin concentration (P=0.01) were higher in progeny from mothers provided with more feed. Body weight at birth was negatively correlated with the abundance of leptin mRNA in subcutaneous fat at 59 days of age (P=0.01). This study shows for the first time that maternal nutrition during pregnancy programs postnatal leptin expression in offspring.

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K. DEMISCH
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J. K. GRANT
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W. BLACK
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SUMMARY

The concentration of testosterone was measured in the plasma of 16 healthy women in late pregnancy and in 14 of these after delivery. The mean testosterone concentration was found to be elevated in late pregnancy, although the range overlapped with the normal. There was a prompt return to normal concentrations after delivery. The significance of the results and of inactivation of testosterone by protein binding are discussed.

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L Thomas
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JM Wallace
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RP Aitken
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JG Mercer
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P Trayhurn
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N Hoggard
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This study examined the pattern of circulating leptin in age-matched sheep during adolescent pregnancy, and its relationship with maternal dietary intake, body composition and tissue expression of the leptin gene. Overfeeding the adolescent pregnant ewe results in rapid maternal growth at the expense of the placenta, leading to growth restriction in the fetus, compared with normal fed controls. Our results demonstrate that, in the adolescent ewe, overfeeding throughout pregnancy was associated with higher maternal leptin concentrations, when compared with moderately fed controls (P<0.05), with no peak in circulating leptin towards the end of pregnancy. There was a close correlation between indices of body composition and circulating leptin levels at day 104 of gestation and at term (P<0.03). Further, when the dietary intake was switched from moderate to high, or high to moderate, at day 50 of gestation, circulating leptin levels changed rapidly, in parallel with the changes in dietary intake. Leptin mRNA levels and leptin protein in perirenal adipose tissue samples, taken at day 128 of gestation, were higher in overfed dams (P<0.04), suggesting that adipose tissue was the source of the increase in circulating leptin in the overnourished ewes. Leptin protein was also detected in placenta but leptin gene expression was negligible. However, leptin receptor gene expression was detected in the ovine placenta, suggesting that the placenta is a target organ for leptin. A negative association existed between maternal circulating leptin and fetal birth weight, placental/cotyledon weight and cotyledon number. In conclusion, in this particular ovine model, hyperleptinaemia was not observed during late pregnancy. Instead, circulating leptin concentrations reflected increased levels of leptin secretion by adipose tissue primarily as a result of the increase in body fat deposition, due to overfeeding. However, there appears to be a direct effect of overfeeding, particularly in the short term. In the nutritional switch-over study, circulating leptin concentrations changed within 48 h of the change in dietary intake. The presence of leptin protein and leptin receptor gene expression in the placenta suggests that leptin could be involved in nutrient partitioning during placental and/or fetal development.

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Cheryl A Conover Division of Endocrinology, Metabolism and Nutrition, Mayo Clinic, 200 First Street SW, 5-194 Joseph, Rochester, Minnesota 55905, USA

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Megan A Mason Division of Endocrinology, Metabolism and Nutrition, Mayo Clinic, 200 First Street SW, 5-194 Joseph, Rochester, Minnesota 55905, USA

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James A Levine Division of Endocrinology, Metabolism and Nutrition, Mayo Clinic, 200 First Street SW, 5-194 Joseph, Rochester, Minnesota 55905, USA

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Colleen M Novak Division of Endocrinology, Metabolism and Nutrition, Mayo Clinic, 200 First Street SW, 5-194 Joseph, Rochester, Minnesota 55905, USA

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Introduction Pregnancy-associated plasma protein-A (PAPP-A) is a recently discovered metalloproteinase whose major physiological function, as so far identified, is to increase local insulin-like growth factor (IGF) bioavailability through the

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M Maggi
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G B Vannelli
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G Fantoni
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E Baldi
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A Magini
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A Peri
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S Giannini
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L Gloria
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P Del Carlo
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D Casparis
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T Tomei
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M Serio
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Abstract

In this study we report the immunolocalization, binding and biological activity of endothelins in the human uterus. Since, in previous studies in the rabbit, sex steroids greatly affected uterine endothelin-1 (ET-1) immunolocalization and binding, we sought to compare results obtained in a relatively steroid-deprived uterus (postmenopausal women) with those obtained in late pregnancy. Two classes of ET receptors were identified in human pregnant and non-pregnant myometrium. One site (ETB) was a low capacity site (0·3 pm/mg protein) that bound with high affinity (0·1 nm), yet no selectivity, ET-1, ET-2, ET-3, sarafotoxin (SRTX) and vasoactive intestinal contractor (VIC). The second site (ETA) was six fold more concentrated than the former (1·9 pm/mg protein) and was relatively selective for ET-1, ET-2 and VIC but showed lower affinity for ET-3 and SRTX. Studies with human myometrial cells indicated that the ETA receptor mediates an increase in intracellular calcium, while the physiological function of the ETB receptor is still unclear. Homologous competition curves for ET-1 were used in order to study the ET receptor density (ETA+ETB) in individual myometrial samples. We found that the concentration of ET receptors did not change during different stages of labour or in postmenopausal women. We identified cells with intense positivity for ET-1 in human decidua. Similar cells were also present in pregnant myometrium, intimately associated with smooth muscle cells. Conversely, no staining for ET-1 was observed in non-pregnant myometrium. A paracrine role for ET-1 in the human uterus is suggested.

Journal of Endocrinology (1994) 142, 385–396

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T K Woodruff
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P Sluss
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E Wang
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I Janssen
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M S Mersol-Barg
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Abstract

Activin A (βA–βA) and activin B (βB–βB) are related dimeric proteins that regulate numerous cellular activities. Activin activity is bioneutralized by follistatin, a specific and high-affinity binding protein. Recently, our group developed specific and sensitive enzyme-linked immunosorbent activin assays that do not detect either activin isoform when bound to follistatin, therefore, the assays are specific for biologically relevant ligands. Activin A is measurable in the serum of pregnant women (cross-sectional sample collection), while activin B is not detected in maternal serum. However, activin B is measurable in amniotic fluid and cord blood sera. The purpose of this study was to measure serum activin A, activin B, and follistatin prospectively in longitudinally collected samples during pregnancy. This study design offered observations of relative changes in serum hormone concentration with each person serving as an internal reference.

Serum samples were collected bimonthly from seven pregnant women beginning within the second month of gestation, and up to, but not including, the onset of labor. Six of the seven women had normal labor and delivery. One patient required pitocin (an oxytocin agonist) for induction of labor which led to delivery. Activin A, activin B, total follistatin, free follistatin, human chorionic gonadotropin, estradiol, progesterone, FSH, and LH were measured in maternal serum samples using specific assays.

Serum activin A levels increased in the final month of pregnancy in the six patients who delivered following normal labor (<0·78 ng/ml (first trimester) to 1–6 ng/ml (term)). Activin B was not detected in any serum sample (<0·78 pg/ml). Total serum follistatin (free follistatin, follistatin–activin, and follistatin–inhibin) increased 10- to 45-fold in the final month of pregnancy in four of the women undergoing normal labor (10 ng/ml (first trimester) to 100–450 ng/ml (final month)). Total follistatin was high and variable in two women throughout pregnancy. Total follistatin returned to basal serum concentration in three of the patients during the last 2 weeks of pregnancy. Free follistatin was detected throughout pregnancy (range <2–35 ng/ml). Free follistatin represented a small percentage of the total follistatin throughout the time of pregnancy and did not rise coincident with the rise in total follistatin. Serum activin A and activin B were not detected during the entire course of pregnancy in the one patient who did not have normal labor and total follistatin did not rise in the last trimester of pregnancy. Gonadotropin and steroid hormones were measured in all patients and were within normative ranges for human pregnancy (inclusive of the non-laboring patient).

The results suggest that immunodetectable activin A is present in the third trimester of pregnant women who have normal onset labor. The total follistatin assay results suggest that follistatin–activin (or –inhibin) complexes are upregulated during the third trimester of pregnancy. Importantly, activin A production exceeds the binding capacity of circulating follistatin. Because binding protein free activin A is biologically active we conclude that the activin A detected in late pregnancy is biologically relevant. The findings are consistent with our hypothesis that activin A is an endocrine factor during the last trimester of human pregnancy and may be involved in normal labor.

Journal of Endocrinology (1997) 152, 167–174

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R. S. Bridges
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R. B. Todd
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C. M. Logue
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Testosterone concentrations in serum of rats bled throughout pregnancy and post partum were measured using Celite microcolumn chromatography and a radioimmunoassay for testosterone. Mean serum levels of testosterone ranged from about 170 to 340 pmol/l during the first 10 days of pregnancy. Significant increases in concentrations of testosterone in serum of pregnant rats were found on days 12, 15 and 18 of gestation. The highest testosterone concentrations occurred on days 18 and 20 of pregnancy when mean levels were 3228 and 3685 pmol/l respectively. Testosterone levels declined before parturition on day 22 (mean = 1449 pmol/l and declined further after parturition (mean = 315 pmol/l). In order to determine whether serum testosterone concentrations varied during the day in the pregnant rat, samples were collected at 6-h intervals on days 6–7 and 14–15 of gestation. Diurnal variations in serum testosterone concentrations were not evident during early or late pregnancy, unlike the rhythmic changes in serum prolactin levels found at these times during early pregnancy. The possible sources of the increased titres of serum testosterone during the second part of gestation in rats are discussed.

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K. BROWN-GRANT
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SUMMARY

Ovarian follicles develop to the stage where they are competent to ovulate in response to exogenous gonadotrophin at the same rate during early pregnancy as in a normal 4-day cycle in the rat. The length of time during which this competency is retained appears to be shorter than after the blockade of ovulation by pentobarbitone administered at pro-oestrus in rats that are not pregnant. Evidence from oviduct:plasma (O:P) and uterus:plasma (U:P) 131I ratio measurements indicating high progesterone and low oestrogen levels at this stage of pregnancy may be relevant to this finding. Some evidence for a 3-day cycle of follicle development after day 6 of pregnancy was obtained but the high O:P ratios suggest that little oestrogen secretion is associated with this.

Oestradiol benzoate (6·25–100 μg.) administered after day 3 of pregnancy will induce ovulation when competent follicles are present. The thyroid:plasma (T:P) concentration ratio for 131I does not rise in association with ovulation induced by exogenous gonadotrophin. After oestrogen administration, the T:P ratio may increase regardless of whether ovulation is induced or not. The results are discussed in relation to the studies of other workers on the induction of ovulation during pregnancy and to earlier work on the relationship between ovulation and changes in thyroid gland activity.

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F. Pansini
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C. M. Bergamini
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M. Malfaccini
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G. Cocilovo
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M. Linciano
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M. Jacobs
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B. Bagni
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ABSTRACT

The distribution of isomorphic forms of prolactin in the serum of pregnant women was studied by gel filtration chromatography. Using this technique we were able to resolve three peaks, detected by radioimmunoassay: they were termed 'big-big', 'big' and 'little' prolactin in order of decreasing size, with approximate molecular weights > 100 000, 50 000 and 21 000 respectively. They displayed a comparable immunoreactivity to the antiserum employed in the radioimmunoassay, as determined in competition experiments. The relative amount of each hormone form in serum changed during the third trimester of pregnancy. At week 33 of pregnancy, 'little' prolactin accounted for 63·2 ± 7·7% of the total circulating hormone present in the serum of five normal pregnant women. During the progression of pregnancy, there was a gradual increase in the low molecular weight prolactin, so that, at the time of delivery, the larger forms of the hormone were present only in small amounts.

J. Endocr. (1985) 106, 81–85

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Y. MANABE
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Mechanical stimulation of the uterus by means of bougies can promote uterine contractility at any time after mid-pregnancy. This principle is commonly used for mid-term abortion as well as for inducing full-term delivery in Japan (Manabe, 1969). Foetuses are normally delivered alive and the placentae are free from any notable histological damage during treatment. In an attempt to assess the possible endocrine control of the onset of labour by this method, the author reported that levels of urinary metabolites of oestrogen and progesterone did not change significantly before and during abortion (Manabe, 1968). The present study was designed to see whether placental progesterone synthesis was maintained at normal levels during bougie-induced abortion in mid-pregnancy. Progesterone levels in peripheral blood were serially estimated during the course of treatment.

Five normal women ranging from the 16th to 27th week of pregnancy were used in this study. One or two sterilized rubber bougies

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