Proposed diagnostic thresholds for gestational diabetes mellitus according to a 75-g oral glucose tolerance test. Maternal and perinatal outcomes in 3260 Danish women.

AIMS: To study if established diagnostic threshold values for gestational diabetes based on a 75-g, 2-h oral glucose tolerance test can be supported by maternal and perinatal outcomes. METHODS: Historical cohort study of 3260 pregnant women examined for gestational diabetes on the basis of risk indicators. Information on oral glucose tolerance test results and clinical outcomes were collected from medical records. RESULTS: There was an increased risk of delivering a macrosomic infant in women with 2-h capillary blood glucose of 7.8-8.9 mmol/l compared with women with 2-h glucose < 7.8 mmol/l. Despite treatment, 2-h glucose of 9.0-11.0 mmol/l and > or = 11.1 mmol/l were both associated with increased rates of macrosomia, spontaneous preterm delivery, hypertensive complications, and neonatal hypoglycaemia. Adverse outcomes tended to be more frequent in women with 2-h glucose > or = 11.1 mmol/l than in women with 2-h glucose of 9.0-11.0 mmol/l. CONCLUSIONS: The risk for several maternal and perinatal complications increased with the diagnostic threshold for 2-h glucose. Large-scale blinded studies are needed to clarify the question of a clinically meaningful diagnosis of gestational diabetes mellitus. Until these results are available, a 2-h threshold level of 9.0 mmol/l after a 75-g oral glucose tolerance test seems acceptable.

Clinical impact of mild carbohydrate intolerance in pregnancy: a study of 2904 nondiabetic Danish women with risk factors for gestational diabetes mellitus.

OBJECTIVE: The objective was to study the clinical impact of mild carbohydrate intolerance in pregnant women with risk factors for gestational diabetes mellitus. STUDY DESIGN: This was a historical cohort study of 2904 pregnant women examined for gestational diabetes on the basis of risk factors. Information on oral glucose tolerance test results and clinical outcomes was collected from laboratory charts and medical records. RESULTS: The following outcomes increased significantly with increasing glucose values during the oral glucose tolerance test: shoulder dystocia, macrosomia, emergency cesarean section, assisted delivery, hypertension, and induction of labor. However, when corrections were made for other risk factors, hypertension and induction of labor were only marginally associated with glucose levels. CONCLUSION: In a group of nondiabetic pregnant women with risk factors for gestational diabetes, there was a graded increase in the frequency of shoulder dystocia and other maternal-fetal complications with increasing glucose levels during an oral glucose tolerance test.

Association between colonization with group B streptococci during pregnancy and preterm delivery among Danish women.

OBJECTIVE: We studied the relationship between group B streptococcal colonization and preterm delivery. STUDY DESIGN: In this prospective study at a single hospital in Odense, Denmark, cervicovaginal cultures were obtained at < or = 24 weeks' gestation from all the women, at delivery from women with preterm deliveries, and from a random sample of women delivering at term. RESULTS: In 2846 singleton births, there was no significant association between group B streptococcal colonization at

Does fetal antigen 1 (FA1) identify cells with regenerative, endocrine and neuroendocrine potentials? A study of FA1 in embryonic, fetal, and placental tissue and in maternal circulation.

Fetal antigen 1 (FA1) is a circulating EGF multidomain glycoprotein. FA1 and its membrane-associated precursor is defined by the mRNAs referred to as delta-like (dlk), preadipocyte factor 1 (pref-1) or zona glomerulosa-specific factor (ZOG). Using a polyclonal antibody recognising both forms, the localisation of FA1/dlk was analysed in embryonic and fetal tissues between week 5 to 25 of gestation and related to germinal origin and development. FA1 was observed in endodermally derived hepatocytes, glandular cells of the pancreas anlage, and in respiratory epithelial cells. FA1 was also present in mesodermally derived cells of the renal proximal tubules, adrenal cortex, Leydig and Hilus cells of the testes and ovaries, fetal chondroblasts, and skeletal myotubes. Ectodermally derived neuro- and adenohypophysial cells, cells in the floor of the 3rd ventricle and plexus choroideus were also FA1 positive. The number of cells expressing FA1 decreased during fetal development where the expression became restricted to specific functional cells. Epidermis, gut epithelium, gall bladder, blood cells, spleen, thyroid gland, salivary glands, and smooth muscle cells were FA1 negative. Analysis of extra-embryonic tissues from normal and pathological pregnancies revealed FA1 in stromal cells surrounding the blood islands of the yolk sac as well as in placental fibroblasts where the expression was most pronounced in diploid, androgenic complete hydatidiform moles. However, as measured by ELISA, the circulating maternal FA1 levels in complete moles were not different from normal pregnancies. The results presented suggest that FA1 is a growth and/or differentiation factor extensively expressed in immature cells and down-regulated during fetal development. FA1 down-regulation was associated with a shift in the subcellular localisation indicating differential post-translational/post-transcriptional modifications during fetal development. FA1 may be a new marker of cellular subtypes with a regenerative potential and of specific cells with endocrine or neuroendocrine functions.

Localization of E-cadherin in villous, extravillous and vascular trophoblasts during intrauterine, ectopic and molar pregnancy.

Previous reports have described down-regulation of E-cadherin in trophoblasts differentiating to an invasive phenotype. This study shows the localization of E-cadherin in a prospective design with stereological sampling of fetal and maternal first, second and third trimester tissue. E-cadherin was observed in villous cytotrophoblasts, and in non-proliferating, intermediate trophoblasts (IT) within cell columns and islands in intrauterine, ectopic and partial molar placentas. Highly proliferating IT with cytological atypia in complete molar placentas were also E-cadherin-positive. E-cadherin was present in trophoblasts throughout the anchoring cell columns. Trophoblasts undergoing epithelial mesenchymal transformation (EMT) detaching from the distal cell columns and deeper located single extravillous interstitial trophoblasts (EVT) showed E-cadherin-negative breaches in the cell membrane. Prior to the late second trimester, the relative number of E-cadherin-positive single EMT and EVT differed from the total number of cytokeratin-positive trophoblasts. Intraluminal, endovascular and perivascular trophoblasts adjacent to the maternal vessels were also E-cadherin-positive, but a highly varying pattern was observed at different ages of gestation. Our results indicate a temporary shift in E-cadherin expression in extravillous trophoblasts possessing a migrating and invasive potential. Functional E-cadherin may be restored as trophoblasts aggregate in the decidua and the vessel wall after completion of migration.

Maternal and perinatal outcomes in 143 Danish women with gestational diabetes mellitus and 143 controls with a similar risk profile.

AIMS: To assess maternal and fetal outcomes in pregnancies complicated by gestational diabetes mellitus (GDM) compared to non-diabetic pregnancies with an otherwise similar risk profile and to study the association between different anti-diabetic treatments and fetal outcomes. METHODS: The records of 143 consecutive GDM pregnancies and 143 non-diabetic controls matched on the basis of age, parity and pre-pregnancy body mass index (BMI) were studied. The GDM patients were treated with diet, tolbutamide and insulin. Data were collected from medical records and birth records. RESULTS: Despite treatment, the GDM group had a statistically significant higher frequency of maternal hypertension (20% vs. 11%), induction of labour (61% vs. 24%), Caesarean section (33% vs. 21%), macrosomia (14% vs. 6%), neonatal hypoglycaemia (24% vs. 0) and admission to a neonatal unit (46% vs. 12%). The risk of complications was similar in the different treatment groups. However, in the tolbutamide-treated group, one case of long-standing severe hypoglycaemia in a premature neonate occurred. CONCLUSIONS: Pregnancies complicated by GDM are associated with a higher frequency of adverse maternal and fetal outcomes. The outcomes seem to be unaffected by treatment modality. However, because of the potential risk of hypoglycaemia in some neonates, tolbutamide treatment cannot be recommended in pregnancy.

[The clinical impact of gestational diabetes mellitus]. / Den kliniske betydning af gestationel diabetes mellitus.

In Denmark, gestational diabetes mellitus (GDM) develops in about 2% of all pregnant women. The discussion of GDM is complicated by lack of consensus regarding screening methods, diagnosis and treatment. Observational studies indicate that untreated GDM is associated with an increased risk of maternal and perinatal morbidity, and that the offspring of GDM mothers tend to be at increased risk of developing diabetes and adiposity as a result of an abnormal intrauterine environment. Several follow-up studies have shown that women with previous GDM run a considerable risk of developing diabetes (especially type 2 diabetes) later in life. Intervention strategies for this high risk group are suggested.

Localization and significance of urokinase plasminogen activator and its receptor in placental tissue from intrauterine, ectopic and molar pregnancies.

Urokinase plasminogen activator receptor (uPAR) is a membrane-anchored protein with urokinase plasminogen activator (uPA) as the ligand. This complex induces proteolysis and remodelling of maternal decidua during placental implantation. The presence of uPAR on trophoblasts is supposed to promote adhesion, migration and invasion. In cancer tissue, high levels of uPAR are correlated with a poor prognosis. This immunohistochemical study shows the localization of uPA and uPAR in a prospective design with stereological sampling of fetal and maternal tissue from normal, ectopic and hydatidiform molar (HM) pregnancies. Cytokeratin and Ki67 were used as markers for trophoblasts and proliferating cells. Membrane-bound uPAR was observed on villous non-proliferating intermediate trophoblasts (IT) within cell columns in intrauterine and ectopic pregnancies. The corresponding proliferating IT with cytological atypia sprouting from the chorionic villi in HM was uPAR-negative. uPA but not uPAR was observed in anchoring distal IT at the attachment-point to the basal plate. In the placental bed, extravillous interstitial trophoblasts were uPA-positive but uPAR-negative. The trophoblast giant cells were both uPA- and uPAR-negative. In relation to the maternal vessels, a focal distribution for uPA and uPAR was present in the endovascular and perivascular trophoblasts. The intraluminal trophoblasts overlying endothelial cells were uPAR-positive only. In maternal tissue from intrauterine and molar pregnancies, uPAR was seen in the decidual cells in a zone facing the anchoring villi and the fibrinoid lesions with embedded trophoblasts. In contrast, the stromal cells of the fallopian tube without a decidual reaction facing the implanted gestation were uPAR-negative. Non-invaded decidual, myometrial and muscular tissue of the pregnant uterus and fallopian tube was extensively positive for uPA whereas 'pseudodecidual' cells from the intrauterine evacuate in patients with an ectopic pregnancy only showed a focal and scanty reaction for uPA. When trophoblast invasion of the decidua was present, the decidual cells were uPA-negative. A semi-quantitative assessment of the receptor was estimated in villous IT within cell columns in normal and molar pregnancies but, in conclusion, quantitative evaluation of uPAR cannot be used to predict development of post-molar persistent trophoblastic disease (PTD).

[The clinical impact of gestational diabetes mellitus]. / Den kliniske betydning af gestationel diabetes mellitus.

In Denmark, gestational diabetes mellitus (GDM) develops in about 2% of all pregnant women. The discussion of GDM is complicated by lack of consensus regarding screening methods, diagnosis and treatment. Observational studies indicate that untreated GDM is associated with an increased risk of maternal and perinatal morbidity, and that the offspring of GDM mothers tend to be at increased risk of developing diabetes and adiposity as a result of an abnormal intrauterine environment. Several follow-up studies have shown that women with previous GDM run a considerable risk of developing diabetes (especially type 2 diabetes) later in life. Intervention strategies for this high risk group are suggested.

Circulating concentrations of placenta protein 14 during the natural menstrual cycle in women significantly reflect endometrial receptivity to implantation and pregnancy during successive assisted reproduction cycles.

Placenta protein 14 (PP14), which is the most abundant product of the secretory endometrium, has been proposed as the best biochemical marker of endometrial function in women. In this study, 19 normogonadotrophic women of infertile couples were monitored with serial measurements of concentrations of PP14, gonadotrophins and sex steroids and ultrasound scanning of endometrial thickness throughout three consecutive cycles. The first two of these were natural, unstimulated cycles (cycles 1 and 2), while ovarian stimulation with clomiphene and human menopausal gonadotrophin combined with assisted reproduction (intrauterine insemination in four cases and in-vitro fertilization in 15) was performed in the third cycle (cycle 3). A newly developed enzyme-linked immunosorbent assay was used to measure serum PP14 concentrations. In cycle 3, seven women became pregnant (group A) and 12 did not (group B). Circulating concentrations of PP14 were significantly lower in group A than in group B throughout all three cycles and in all cycle phases with exception of the late luteal phase of cycle 3, during which PP14 concentrations in group A were significantly higher than in group B. Statistical analyses showed no significant correlations between serum concentrations of PP14 and follicle stimulating hormone, luteinizing hormone and progesterone, and endometrial thickness. By contrast, serum oestradiol concentrations during the pre-ovulatory phase were significantly correlated with PP14 concentrations during the mid-luteal phase of the cycle. It is concluded that circulating PP14 is a most reliable biochemical marker of endometrial function in women and that relatively low concentrations in serum during the natural, unstimulated cycle are significantly correlated to implantation and pregnancy during successive assisted reproduction cycles. Measurement of PP14 in serum may thus be useful as a method of screening endometrial function in women, before commencing troublesome and costly treatment for infertility. However, further studies in a much larger number of women are needed to confirm this observation and to elucidate the as yet undefined physiological functions of PP14 in women.

Ovarian volume in gynecologically healthy women using no contraception, or using IUD or oral contraception.

OBJECTIVE: The aim of this study was to determine the ovarian volume by transvaginal ultrasonography in a gynecologically healthy population of women using no contraception, using intrauterine contraceptive device, or using oral contraceptive. MATERIALS AND METHOD: The study had a cross-sectional design. The ovaries of 428 women aged 1445 who contacted the family planning clinic in the county of Funen were examined. Most of the statistical analyses were carried out using standard techniques. However polynominal regression analysis was used to model ovarian volumes as a function of the day of cycle. RESULTS: No differences between the volumes of the right and the left ovary were found in any of the groups. Significant differences were found between the ovarian volumes of the three groups. The ovarian volumes were found to be largest in women using intrauterine contraceptive device, lesser in women using no contraception and smallest in women using oral contraception. A significant difference was found of the ovarian volumes throughout the menstrual cycle in women who were not using oral contraception. The ovarian volumes did not change throughout the menstrual cycle in women using oral contraception. In women not using oral contraception the largest ovary increased in volume from the start of the cycle to day 19, thereafter the volume declined. No evidence of any change of volume over the menstrual cycle was found in the smallest ovary and, for women using oral contraception, both ovaries. There was no correlation between age, height, weight, parity, and ovarian volume in any of the groups. CONCLUSION: The ovarian volumes, in gynecologically healthy women using intrauterine contraceptive device, are larger than in women using no contraception. It appears that oral contraception reduces the volumes of both ovaries in all phases of the menstrual cycle to equal levels.

PIP: The aim of this cross-sectional study was to determine the ovarian volume by transvaginal ultrasonography in a gynecologically healthy population of women using no contraception, using IUDs, or using oral contraceptives (OCs). The ovaries of 428 women aged 14-45 who contacted the family planning clinic in the county of Funen were examined. Most of the statistical analyses were carried out using standard techniques. However, polynominal regression analysis was used to model ovarian volumes as a function of the day of cycle. No differences between the volumes of the right and the left ovary were found in any of the groups. Significant differences were found between the ovarian volumes of the three groups. The ovarian volumes were found to be largest in women using IUDs, lesser in women using no contraception, and smallest in women using OCs. A significant difference was found of the ovarian volumes throughout the menstrual cycle in women who were not using OCs. The ovarian volumes did not change throughout the menstrual cycle in women using OCs. In women not using OCs the largest ovary increased in volume from the start of the cycle to day 19; thereafter the volume declined. No evidence of any change of volume over the menstrual cycle was found in the smallest ovary and, for women using OCs, in both ovaries. There was no correlation between age, height, weight, parity, and ovarian volume in any of the groups. The ovarian volumes in gynecologically healthy women using IUDs are larger than in women using no contraception. It appears that OC use reduces the volumes of both ovaries in all phases of the menstrual cycle to equal levels.

Circulating levels of relaxin are normal in pregnant women with pelvic pain.

OBJECTIVE: The hormone relaxin induces loosening of the pelvic ligaments and joints in several species. Previous studies have suggested a similar role for relaxin during human pregnancy. Furthermore, a correlation has been noted between high circulating levels of this hormone and severe pelvic pain in pregnant women. The present study was designed to evaluate whether serum relaxin concentrations were elevated in pregnant women with clear subjective and objective evidence of pain attributable to relaxation of the pelvic ligaments. STUDY DESIGN: Serum relaxin was measured at week 33 of gestation in 455 pregnant women with clearly defined pain in their pelvic joints and 455 normal pregnant controls matched for age and parity. All participants underwent an examination consisting of a structured questionnaire and fifteen specific tests for pelvic joint pain. The group with pain was further subdivided into four subgroups with different levels of disability and prognosis. Relaxin concentrations were measured using enzyme-linked immunoassay. RESULTS: There was no difference in serum relaxin concentration between the control and study group, nor between the subgroups of women with pelvic pain. CONCLUSION: We failed to confirm an earlier claim that circulating relaxin levels are related to pelvic girdle pain in pregnant women.

Changes in blood levels of proteinase inhibitors, pregnancy zone protein, steroid carriers and complement factors induced by oral contraceptives.

Three low-dose oral contraceptives Trinordiol, Gynatrol, and Marvelon, containing ethinylestradiol (EE) in combination with triphasic levonorgestrel (LNg), monophasic levonorgestrel, and monophasic desogestrel (DGS), respectively, were given to 65 healthy women, n = 21-22 in each group. Blood levels of antithrombin III (AT III), alpha 2-macroglobulin (alpha 2M) alpha 1-antitrypsin (alpha 1at), complement factors (factor B, C3, C4), pregnancy zone protein (PZP), corticosteroid binding globulin (CBG), sex hormone binding globulin (SHBG) and albumin were measured before treatment and during the first and third treatment cycles. AT III levels decreased and alpha 1at levels increased in all three groups during treatment. alpha 2M increased during cycle 3 in the Trinordiol and Gynatrol groups. CBG, PZP and SHBG levels increased in all 3 groups, the CBG and PZP increase being higher in the Marvelon group than in the Gynatrol group. Increases in SHBG levels were found in the order Marvelon > Trinordiol > Gynatrol. Plasma levels of complement factors B, C3 and C4 remained unchanged. It is concluded that the increase in alpha 1at partly compensates for the fall in AT III, that the rise in PZP presumably enhances fibrinolysis, and that LNg has higher anti-estrogenicity and androgenicity than DSG.

PIP: Clinicians administered 3 types of oral contraceptives (OCs) to 65 healthy 15-31 year old women attending the Public Family Planning Clinic in Odense, Denmark, to determine the effect of the 3 OCs on enzyme inhibitors, complement factors, and steroid dependent proteins. The OCs were a low-dose monophasic containing ethinyl estradiol (EE) and desogestrel (Marvelon), another low dose monophasic containing EE and levonorgestrel (Gynatrol), and a triphasic containing EE and levonorgestrel (Trinordiol). 21-22 women were assigned to each group. Pretreatment values of the biochemical parameters were not significantly different between the 3 groups. In each group, mean plasma antithrombin III (AT III) levels fell (p .04-.002) and alpha1-antitrypsin (alpha1-at) levels increased (p .0001). Mean plasma levels of alpha2-macroglobulin rose sharply during cycle 3 in the Trinordiol and Gynatrol groups (p .04 and .002, respectively). In all 3 groups, mean serum levels of corticosteroid binding globulin (CBG), pregnancy zone protein (PZP), and sex hormone binding globulin (SHBG) increased significantly (p .03-.0001). The CBG and PZP levels were higher in the women using Marvelon than in those using Gynatrol. The increase in SHBG levels were only significant in the Trinordiol and Marvelon groups, however (p .0001). Mean plasma levels of complement factors B, C3, and C4 and of albumin did not change. Based on these results, the researchers concluded that the increase in alpha1-at only partially offsets the decrease in AT III, that the increase in PZP may intensify fibrinolysis, and that levonorgestrel has greater antiestrogenicity and androgenicity than desogestrel.

Changes induced in serum protein profiles by ovarian stimulation during in-vitro fertilization--embryo transfer treatment: a comparison between conception and non-conception cycles.

The profiles of plasma protein concentrations during the follicular phase in unstimulated women and in women undergoing ovarian stimulation for in-vitro fertilization--embryo transfer (IVF-ET) treatment are described. Plasma protein concentrations are correlated with those of total oestradiol (protein-bound and free) and total progesterone. In addition, 10 conception cycles and 18 non-conception cycles are compared in an attempt to identify predictors of successful treatment. Ovarian stimulation caused a significant increase in follicular phase in serum concentrations of sex hormone binding globulin (SHBG), cortisol binding protein (CBP) and insulin-like growth factor binding protein 1 (IGFBP1). In contrast no increase was observed in unstimulated cycles. Serum levels of endometrial protein PP14 decreased significantly during the follicular phase in both stimulated and unstimulated cycles. Levels of pregnancy zone protein (PZP) were more than doubled at the time of oocyte aspiration compared to the unstimulated cycles. Albumin concentrations were unchanged by the stimulation. Throughout the follicular phase, levels of SHBG were significantly higher, and total oestradiol significantly lower in women who became pregnant, than in those who did not. Therefore, a low concentration of free, biologically active oestradiol seemed to favour pregnancy, as the concentration of albumin is similar in the two groups. The endometrial protein PP14 was significantly lower during the follicular phase in conception than in non-conceptional cycles. On day 2 of the treatment cycle, the PP14 concentration showed a 75% correct prediction of conception and non-conception cycles. These results suggest that levels of PP14 may predict successful IVF cycles even before hormonal treatment is commenced.

Open randomized comparison of prostaglandin E2 given by intracervical gel or vagitory for preinduction cervical ripening and induction of labor.

Intracervical application of prostaglandin E2 (PGE2) in a viscous gel was compared with conventional wax-based PGE2 vagitories (pessaries) for ripening of the cervix prior to induction of labor. A total of 226 healthy pregnant women at term were randomly allocated to receive intracervical gel with an effective dose of 0.5 mg (n = 116) or vagitories containing 2.5 mg PGE2 (n = 110). All women had a modified cervical score of less than or equal to 4. The numbers of cases contributed by each of the three centers were similar. There was no significant difference in parity, gestational length, maternal characteristics, indications for induction or preinduction cervical scores between the treatment groups. The rate of spontaneous birth was 71% in the gel group, compared with 69% in the vagitory group. Successful treatment was defined as active labor within 24 h or a change in cervical dilatation allowing artificial rupture of the membranes with subsequent progressive labor. The success rate was not significantly different in the gel group (82%) compared with the vagitory group (80%). There were no differences in the frequency of fetal distress, outcome of labor, assisted delivery rates or maternal side effects. The cervical scores were not different at 12 and 24 h after application. Intracervical gel and intravaginal application of PGE2 were similar in their efficacy and safety for ripening of the cervix and inducing labor at term.

A biochemical test for the direct assessment of endometrial function: measurement of the major secretory endometrial protein PP14 in serum during menstruation in relation to ovulation and luteal function.

Circulating PP14 was measured by radioimmunoassay in ovulating (n = 12) and anovulatory (n = 3) women throughout the menstrual cycle, the highest levels of serum PP14 being seen during menstruation and in the late luteal phase in ovulating women. Mean serum PP14 levels on days 1-7 and 24-28 of the menstrual cycle were significantly higher than those observed from days 8 to 23 (P less than 0.0005 and P = 0.005 respectively). There was no difference in mean PP14 levels observed in the menstrual and luteal phase. By contrast, serum PP14 was rarely detected in anovulatory cycles. During the luteal phase, mean serum PP14 levels were apparently not related to serum progesterone levels. However, mean PP14 levels during the menstrual phase were significantly higher in the group of women with the highest progesterone production (Pmax greater than 39 nmol/l) (P less than 0.002) in comparison with levels seen in ovulating women with lower progesterone production (Pmax less than 32 nmol/l). These findings suggest that the synthesis and secretion of PP14 is influenced by ovulation and luteal function. Serum PP14 measurements may provide useful information about the endometrium in relation to fertility, and that these measurements during the menstrual cycle may distinguish between ovulatory and anovulatory cycles.

De novo synthesis of placental protein-14 (PP14) and not PP12 by monolayer cultures of glandular epithelium of gestational endometrium.

Nine monolayer cell cultures of glandular epithelium from gestational endometrium were established from six apparently healthy women undergoing elective termination of pregnancy (7-11 weeks gestation). Radiolabel incorporation studies showed increasing incorporation of [35S]methionine into proteins present in supernatant and cytosol fractions over 48 h. The secreted proteins represented approximately 20% of the total incorporation of methionine into cytosolic proteins. De novo synthesis and secretion of placental protein-14 (PP14) and not PP12 was identified by a novel combination of line immunoelectrophoresis and autoradiography. All monolayer cultures demonstrated the presence of radiolabeled PP14, but not PP12, in the culture supernatants. These observations suggest that the glandular epithelial cells are the major site of synthesis and secretion of PP14 in human gestational endometrium.

Identification of specific serum proteins synthesized de novo by monolayer cultures of glandular cells of gestational endometrium.

Monolayer cell cultures (n = 3) of glandular epithelium of gestational endometrium obtained from three apparently healthy women undergoing elective termination of pregnancy (7-9 weeks gestation) were established. De novo synthesis of eight serum proteins (albumin, alpha 1-antitrypsin, ceruloplasmin, beta-lipoprotein, alpha 2-macroglobulin, fibronectin and complement factors C3 and C4) was demonstrated by the incorporation of radiolabelled substrate ([35S]methionine) employing autoradiography (AR) in combination with crossed immunoelectrophoresis (XIE), referred to as ARXIE, and line immunoelectrophoresis (LIE), referred to as ARLIE. By contrast, there was no evidence for de novo synthesis of IgA, haptoglobin and orosomucoid. Our findings suggest that the gestational endometrium may contribute to the production of several proteins considered to be synthesized and secreted mainly by the liver and reticulo-endothelial system. The simple techniques used here to identify the de novo synthesis of human serum proteins could be applied to investigate protein synthesis by a wide range of tissues and cells.

Two fetal antigens (FA-1 and FA-2) and endometrial proteins (PP12 and PP14) isolated from amniotic fluid: localisation in the fetus and adult female genital tract.

Monospecific antisera against two fetal antigens (FA-1 and FA-2), alphafetoprotein (AFP) and two endometrial proteins (PP12 and PP14) were used to examine the distribution of these proteins and antigens in human trophoblast and gestational endometrium in first and third trimesters of pregnancy, normal human ovary and fetal tissues by indirect immunoperoxidase histochemical localisation techniques. Fetal liver stained exclusively for FA-1 and AFP which was used as a reference protein. Staining for FA-2 was seen in fetal connective tissue, in particular the basement membrane. FA-1 and FA-2 did not stain positively in decidua, trophoblast or ovarian tissue. Gestational endometrium stained positively for PP14 exclusively in the glandular epithelium, whilst staining for PP12 was seen only in the stromal cells. Trophoblast, both early and late, and ovarian tissue did not stain positively for any of the four substances tested.