We studied the in-vitro secretory function of non-polarized and polarized cultured Fallopian tube epithelial cells by measurement of the placental protein 14 (PP14) secretion in primary cultures and subcultures from Fallopian tubes obtained from eight premenopausal women in different phases of the ovarian cycle. Primary cultures were established in minimal essential medium in Earle's salts supplemented with fetal bovine serum and the cells were subcultured for six passages, in the polarized cell cultures, the cells being seeded on an extracellular matrix system. Cell freezing was carried out using 10% dimethyl sulphoxide. PP14 secretion into the culture media was measured by a radioimmunoassay using 125I-PP14 as label and rabbit anti-human PP14 serum. There was a large amount of PP14 secretion into the culture media in primary cultures, the secretion decreasing considerably after subculture 1. PP14 secretion after subculture 2 was not different from the control values. Polarized and non-polarized cells secreted similar amounts of PP14 and frozen-thawed cells did not appear to secrete PP14. Epithelial cells from Fallopian tubes obtained at different phases of the ovarian cycle did not appear to show any difference in PP14 secretion rates. Our data suggest that the in-vitro secretion of PP14 by human Fallopian tube epithelial cells is adversely affected by cell ageing and freezing.

The oviduct has many roles in the process of natural conception, beginning with retrieval of the oocyte released by the ruptured ovarian follicle, and facilitation of the final maturation of sperm which have passed through the uterus (Adams and Chang 1962; Bedford 1968; Hunter and Hall 1974). Both male and female gametes are transported in the oviduct, and meet within well-defined time limits which are species-dependent (Andersen 1927; Harper 1961a,b; Blandau 1969; Boling 1969; Hunter 1974; Blandau and Verdugo 1976; Croxatto and Ortiz 1975; Pauerstein 1975; Eddy et al. 1976). Fertilisation takes place at the isthmic-ampullary junction and the pre-embryo is then transported to the uterine cavity at the optimum time for nidation. While in the oviduct, early development of the mouse pre-embryo appears to be enhanced (Bowman and McLaren 1970; Harlow and Quinn 1982). This may also be the case in humans and has led to studies to determine whether it is beneficial for pre-embryos to be cocultured with oviduct epithelial cell cultures (Bongso et al. 1990). During reproductive events in the oviduct the male gametes and pre-embryo, which are antigenically different from the female host, do not appear to be affected by the maternal immune system. The mechanism of this is unknown.

Prenatal DiagnosisVolume 14, Issue 9 p. 899-900 Letters to the Editor Pregnancy-associated plasma protein A (PAPP-A), a first-trimester screening test for Down syndrome and other chromosomal anomalies B. Brambati, B. Brambati Institute of Obstetrics and Gynaecology, University of Milan, Milan, ItalySearch for more papers by this authorM. C. M. Macintosh, M. C. M. Macintosh Departments of Reproductive Physiology, and Obstetrics and Gynaecology, St Bartholomew's Hospital Medical College and the London Hospital Medical College, London, U.K.Search for more papers by this authorK. Shrimanker, K. Shrimanker Departments of Reproductive Physiology, and Obstetrics and Gynaecology, St Bartholomew's Hospital Medical College and the London Hospital Medical College, London, U.K.Search for more papers by this authorT. Chard, T. Chard Departments of Reproductive Physiology, and Obstetrics and Gynaecology, St Bartholomew's Hospital Medical College and the London Hospital Medical College, London, U.K.Search for more papers by this authorJ. G. Grudzinskas, J. G. Grudzinskas Departments of Reproductive Physiology, and Obstetrics and Gynaecology, St Bartholomew's Hospital Medical College and the London Hospital Medical College, London, U.K.Search for more papers by this author B. Brambati, B. Brambati Institute of Obstetrics and Gynaecology, University of Milan, Milan, ItalySearch for more papers by this authorM. C. M. Macintosh, M. C. M. Macintosh Departments of Reproductive Physiology, and Obstetrics and Gynaecology, St Bartholomew's Hospital Medical College and the London Hospital Medical College, London, U.K.Search for more papers by this authorK. Shrimanker, K. Shrimanker Departments of Reproductive Physiology, and Obstetrics and Gynaecology, St Bartholomew's Hospital Medical College and the London Hospital Medical College, London, U.K.Search for more papers by this authorT. Chard, T. Chard Departments of Reproductive Physiology, and Obstetrics and Gynaecology, St Bartholomew's Hospital Medical College and the London Hospital Medical College, London, U.K.Search for more papers by this authorJ. G. Grudzinskas, J. G. Grudzinskas Departments of Reproductive Physiology, and Obstetrics and Gynaecology, St Bartholomew's Hospital Medical College and the London Hospital Medical College, London, U.K.Search for more papers by this author First published: September 1994 https://doi.org/10.1002/pd.1970140925Citations: 2AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat No abstract is available for this article.Citing Literature Volume14, Issue9September 1994Pages 899-900 RelatedInformation

Serum measurements of pregnancy‐associated plasma protein A (PAPP‐A) and the free β‐human chorionic gonadotrophin (hCG) subunit were made in 13 women with Down syndrome (DS) pregnancies and six other women with fetal aneuploidy ascertained at chorionic villus sampling (CVS), as well as 89 women with contemporaneous normal control pregnancies. Median serum PAPP‐A measurements (0·31 MOM, 95 per cent confidence interval (CI) 0·22–0·65 vs. normal 1·06, 95 per cent CI 0·89–1·20) were lower and free β‐hCG subunit measurements (1·13 MOM, 95 per cent CI 0·93–2·63 vs. normal 0·91, 95 per cent CI 0·79–1·03) were higher at statistically significant levels. Receiver operator characteristic (ROC) curves showed that the highest sensitivity for detection, 71·2 per cent (95 per cent CI 54·7–87·6 per cent), was for depressed PAPP‐A levels; the combination of low serum PAPP‐A levels, maternal age, and elevated free β‐hCG levels yielded a detection rate of 78·9 per cent (95 per cent CI 64·9–92·8 per cent) of the affected pregnancies at 8–12 weeks' gestation.

De-novo synthesis and secretion of protein by short term explants of mucosa from each anatomical section of the Fallopian tube and endometrium of pre-menopausal (n = 25) and tubal mucosa of post-menopausal (n = 5) women were studied by demonstration of incorporation of radiolabelled L-[35S]methionine and one-dimensional SDS — polyacrylamide gel electrophoresis. A consistent finding in 25 pre-menopausal women was the presence of a 25 kDa protein band synthesized by tissue obtained throughout the ovarian cycle. Western blotting demonstrated that this protein band contained placental protein 14 (PP14)-like immunoreactivity in the proliferative (n = 2) and luteal phase (n = 2) of the ovarian cycle. To determine if there is quantitative variation in total protein and PP14 synthesis and secretion during the ovarian cycle, the total quantities of protein and PP14 synthesized were determined by Coomassie Brilliant Blue staining and radioimmunoassay respectively. Analysis of the results of total protein assay revealed statistically significant differences in relation to the anatomical origin of the study tissue (P < 0.01), the stage of the ovarian cycle (P < 0.04) and the manner in which each anatomical site varied during the ovarian cycle (P < 0.01), the endometrium being significantly different from the Fallopian tube. When the data for PP14 synthesized by the Fallopian tube mucosa were analysed, these effects were not seen. PP14 was not detected in the culture media of Fallopian tube mucosa obtained from post-menopausal women.

Objective To assess the relation between maternal serum pregnancy associated plasma protein A (PAPP-A) in the first trimester and the outcome of pregnancy by karyotype.Design A retrospective study of PAPP-A levels in blood samples collected prior to chorionic villus sampling.Setting Milan, Italy.Subjects Five hundred twenty-two women, aged 20 to 47, at 7 to 11 weeks gestation, prior to undergoing chorionic villus sampling. Four hundred forty-five women had a pregnancy with a normal karyotype; in 30 pregnancies the karyotype was abnormal (including 14 cases of Down's syndrome and 7 of trisomy 18).Main outcome measures Normal or abnormal fetal karyotype. Serum PAPP-A at 6 to 11 weeks gestation measured by radioimmunoassay.Results The median value of PAPP-A in the abnormal group was 0.27 multiples of the normal median (MoM). This is significantly lower than the median value in the normal group (1.01 MoM) (95% CI for the difference 0.46-0.84 MoM; P<0.00001 Mann-Whitney test).Conclusions There is an association between low levels of PAPP-A in the first trimester with chromosome anomalies. Screening by measurement of PAPP-A might detect 60% of cases of Down's syndrome in the first trimester with a false positive rate of 5%.

De-novo synthesis and secretion of proteins by short-term explants of matched Fallopian tube mucosa and endometrium were studied using radiolabelled L-[35S]methionine and [3H]glucosamine. To compare directly each anatomical site of the Fallopian tube and endometrium from the same source, newly synthesized proteins were separated on one-dimensional polyacrylamide gel electrophoresis and examined by auto-radiography. De-novo synthesis of two protein bands provisionally designated as tubal epithelial protein 1 (TEP-1) and tubal epithelial protein 2 (TEP-2), was observed in explants of the Fallopian tube mucosa obtained from each anatomical site throughout the ovarian cycle (n = 20). TEP-2 was not apparent in tubal mucosa obtained from post-menopausal women (n = 5). De-novo synthesis of TEP-1 and TEP-2 was not apparent in autoradiographs of radiolabelled proteins from short-term explants of endometrium. From the autoradiographs the molecular mass of TEP-1 and TEP-2 was calculated to be 25 kDa and 17 kDa, respectively. Incorporation of glucosamine into newly synthesized protein occurred in TEP-2 but not TEP-1. TEP-1 was observed to be immunochemically identical to immunoglobulin kappa light chains.

Levels of norethisterone and medroxyprogesterone acetate were measured in serum and milk of women receiving the injectable contraceptive formulations Norigest and DepoProvera, respectively, throughout complete injection intervals. In 5 of 10 women receiving Norigest, serum norethisterone levels were undetectable by 8 weeks after injection and only 2 women had detectable levels of norethisterone in milk at this time. In contrast, 8 of 10 women receiving DepoProvera had detectable levels of medroxyprogesterone acetate in both serum and milk 12 weeks after injection. The ratio of the milk:serum concentrations of norethisterone varied from 0.12 to 0.92 (mean 0.34) and for medroxyprogesterone acetate from 0.12 to 2.60 (mean 0.88). It is unlikely that these differences between the two formulations are due entirely to differences between the binding of norethisterone and medroxyprogesterone acetate to serum proteins. The area under the curve of serum steroid concentrations plotted against time was only 50% higher for women injected with DepoProvera than for those injected with Norigest but the area under the curve for milk values was 400 times higher. Assuming the infant ingests 600 ml milk daily, the daily intake of steroids in the first week after injection would be 0.5 to 2.4 μg for norethisterone and 1 to 13 μg for medroxyprogesterone acetate. By 8 weeks after injection, the amount of norethisterone ingested would be small but that of medroxyprogesterone acetate would still be significant.

A very sensitive competitive protein binding assay for biotinyloestrone. using competition between biotinyloestrone conjugate and 125I-labelled biocytin for avidin is described. The method is based upon the fact that the chemical modification or conjugation to the carboxyl moiety of biotin does not substantially alter the very high Ka of the protein avidin binding its natural ligand biotin.

The bioavailability of lynestrenol and ethynyloestradiol from a capsule formulation was superior to a normal tablet formulation. Bioavailability, as measured by the area under the plasma concentration curves, showed, particularly in the case of the capsule formulation, that all of the administered lynestrenol was converted to norethisterone.

2 populations attending WHO centers, one in Sweden and one in India, participated in a comparative, pilot trial of 2 increasingly popular injectable progestin-only female contraceptives, Depo-Provera and Norigest. The purpose of the study was to assess the pharmacokinetic and pharmacodynamic properties of the 2 formulations (depot medroxyprogesterone acetate and norethisterone enanthate). Differences were found between Swedish women and Indian women in their reactions to the 2 drugs: 1) Norigest was detectable in blood samples a significantly shorter time after injection of the agent in Indian women than in Swedish women; this difference was not apparent with Depo-Provera. 2) Although there was no difference at the 2 centers in the time of ovulation return for subjects receiving Norigest, 0 of 4 Swedish women ovulated more than 156 days after Depo-Provera injection, whereas all 4 Indian women ovulated within 73 days of Depo-Provera injection; in the Swedish women, the levels of medroxyprogesterone were undetectable at time of return to ovulation, whereas Indian women had levels of .6 ng/ml when ovulation resumed. 3) In both cultures, Depo-Provera users had significantly more episodes of bleeding and spotting than Norigest users. This preliminary report emphasizes the variety of responses possible to injection of different contraceptive progestins among various populations and points to the need for further culturally comparative studies.

The concentration of norethisterone in plasma samples from subjects receiving I mg of the steroid orally was measured by radioimmunoassay without chromatography and by gas chromatography-mass fragmentography (g.c.-m.f.). Although values obtained by radioimmunoassay were usually higher than those obtained by m.f. the correlation coefficient for the two sets of results was 0.96. For concentrations of norethisterone above 500 pg/ml the mean over-estimate by radioimmunoassay was less than 30%.

Mean levels of MPA in blood taken 90 days after injection were not significantly different between women who had received a single injection of DepoPrevera and those who had received eight injections at 90-day intervals. There was a wide variation in plasma MPA concentration in the two groups of women. In women who had received 31 to 45 injections of CycloProvera, plasma levels of MPA 28 days after injection were significantly higher than those of women who had received a single injection. The levels were also higher than those found in women 90 days after injection of DepoProvera. The results suggest that the dose of MPA in CycloProvera could be reduced.

The rate of metabolism of orally administered norethisterone was compared in fourteen centres by measuring plasma levels of the steroid by radioimmunoassay at varying times after oral administration of a 1 mg dose. The inter-centre differences were of the same order as the intra-centre differences. Variations in metabolism appeared not to be due to variations in body size.

The duration of action of long-acting injectable formulations of gestagens was assessed in rabbits by measuring blood levels of the gestagen. Norethisterone oenanthate was rapidly absorbed and metabolised and norethisterone was detectable in plasma for up to 60 to 70 days. After administration of Depo-Provera, plasma levels of MPA were detectable for up to 110 days and after injection of norgestrel pelargonate, norgestrel was detected in plasma for a similar period of time. Org 2154 was slowly metabolised and plasma levels of the free gestagen were detectable for more than l80 days. The order of duration of activity is compared with results obtained in baboons and humans. The rabbit may serve as a suitable animal model for predicting the duration of action of injectable formulations in humans.

A radioimmunoassay for measuring medroxyprogesterone acetate in serum or plasma is described using an antiserum produced against the steroid-3-carboxymethyloxime coupled to bovine serum albumin. The method has adequate accuracy, precision and sensitivity and the reliability criteria of the method are compared with those of other radioimmunoassays described for this steroid. Seven days after the intramuscular injection of 150mg Depo-Provera, serum levels of medroxyprogesterone acetate ranged from 1750–9000 pg/ml and by 75 days these levels had decreased to 680–2600 pg/ml.

Ovarian function was studied in ten women receiving a single intramuscular injection of 200 mg norethisterone oenanthate. Two of the ten women appeared to ovulate within 60 days of injection and a further four within 90 days. Follicular activity probably not followed by ovulation occurred in five subjects within 60 days of the injection and in a further subject within 90 days. Only one subject showed no evidence of follicular or luteal activity during the course of this study. There was wide variation in the rate at which the administered gestagen was metabolised and no correlation between the rate of metabolism of norethisterone and the return of ovarian function or the ponderal index, height or weight of the subjects.