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PubMed1993

In-vitro synthesis of total protein and placental protein PP14 by the fallopian tube mucosa: variation in relation to anatomical site, the ovarian cycle and the menopause.

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.

Adult↗
PubMed1993

The production of placental protein 14 and interleukin 6 by human endometrial cells in culture.

Epithelial and stromal cells prepared from endometrium taken at different times in the menstrual cycle were grown in primary culture and the production of placental protein 14 (PP14) and interleukin-6 (IL6) measured. Only the epithelial cells produced PP14. Epithelial cells from endometrium in the late secretory phase produced significantly greater amounts of PP14 (42 +/- 5.8 ng/24 h) compared with that produced by cells from early secretory and proliferative endometrium (16 +/- 1.7 and 11 +/- 1.9 ng/24 h respectively). PP14 production by cells from endometrium at all stages in the cycle was increased by progesterone, or progesterone and oestradiol together, while oestradiol alone had no effect on PP14 production. The greatest stimulation was seen during the early secretory phase when stimulated levels of PP14 reached those obtained during the late secretory phase. IL6 production by epithelial cells also varied depending on the phase of the menstrual cycle. More IL6 was produced by the cells prepared from the endometrium in the proliferative phase (10.9 +/- 0.56 ng/24 h) compared with that produced by cells from early and late secretory endometrium (2.5 +/- 0.19 and 1.45 +/- 0.09 ng/24 h respectively). Addition of steroids to the media stimulated the production of IL6 by cells from proliferative and early secretory endometrium but decreased IL6 production from cells in the late secretory phase. IL6 was also produced by stromal cells but could only be detected in supernatants of cells prepared from late secretory endometrium, and the amounts produced (0.8 +/- 0.09 ng/24 h) were less than that produced by epithelial cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1994

Serum placental protein 14 concentrations are similar in the first trimester pregnancies of women after pituitary down-regulation with a gonadotrophin-releasing hormone agonist and normal cycles with frozen embryo transfers.

Previous studies suggest that, in pregnancies after in-vitro fertilization (IVF) and embryo transfer following pituitary down-regulation with a gonadotrophin-releasing hormone analogue (buserelin) and ovulation induction with human gonadotrophins, the serum placental protein 14 (PP14) concentration is lower than in normally conceived pregnancies. We studied serum PP14 concentrations in two groups of women: (i) in 17 infertile women whose pregnancy followed IVF and embryo transfer using buserelin (long protocol) and human menopausal gonadotrophin for ovulation induction; (ii) in 15 women whose pregnancy followed transfer of frozen-thawed embryos. Similar PP14 concentrations were found in both groups on days 9-10, 14-15 and 70-77 after human chorionic gonadotrophin administration (buserelin, IVF/embryo transfer) or spontaneous luteinizing hormone surge (frozen-thawed embryo transfer). Our results show that PP14 secretion is not compromised by pituitary down-regulation with buserelin in infertile women with functional ovaries.

Buserelin↗
PubMed1994

Placental protein 14 in cycles with normal and retarded endometrial differentiation.

The purpose of this study was to investigate whether endometrium with retarded development differs, functionally, from endometrium with normal 'in-phase' development. Precisely timed endometrial biopsies were obtained from 24 women suffering from unexplained infertility at 4, 7, 10 and 13 days following the luteinizing hormone (LH) surge. Frozen sections were labelled with an anti-placental protein (PP) 14 monoclonal antibody using an avidin-biotin peroxidase technique and semi-quantification of endometrial PP14 was performed using a Quantimet 970 image analyser. Serum PP14 and saliva progesterone were measured for each patient. Data were analysed using one- and two-way analysis of variance. Normal and retarded endometrium were identified in 16 (group I) and eight (group II) women respectively. Both groups demonstrated a significant increase of the area of precipitate measured for PP14 from day LH + 4 to LH + 13. However, two-way analysis of variance showed that endometrial PP14 was significantly (P < 0.05) lower in the retarded endometrium group at LH + 10 and LH + 13. Serum PP14 was also significantly lower (P < 0.01) in women with retarded endometrial development at LH + 13. Women with normal endometrial development had a significantly higher (P < 0.05) concentration of cumulative saliva progesterone from LH + 3 to LH + 5. This study indicates that there are functional differences between normal and retarded endometrium. These differences may adversely affect uterine receptivity during implantation and the early placentation stage.

Adult↗
PubMed1994

Uterine endocrinology and paracrinology: insulin-like growth factor binding protein-1 and placental protein 14 revisited.

A large number of proteins and peptides have been identified in the endometrium where they are likely to exert local biological effects. These substances may be enzymes, their inhibitors, proteinases, proteinase inhibitors, hormones, or bioactive peptides with diverse functions. Endometrial function and embryo-endometrial interactions require synchronized actions between endocrine and local factors. As examples of local factors recent studies on the insulin-like growth factor (IGF) system and placental protein 14 (PP14) are reviewed. IGF binding protein-1 (IGFBP-1) and PP14 are products of the secretory phase endometrium: IGFBP-1 is produced by decidualized stromal cells and PP14 by glandular epithelial cells. IGFBP-1 may inhibit the action of IGFs at the endometrial-trophoblastic interphase, and it may also have a role in stromal-epithelial interaction. PP14 has immunosuppressive properties, and recent findings indicate that it may play a part in the fertilization process by inhibiting binding of spermatozoa to the zona.

Carrier Proteins↗
PubMed1994

Stimulation of human endometrial epithelial cell interleukin 6 production by interleukin 1 and placental protein 14.

The effect of interleukin 1 (IL1) and placental protein 14 (PP14) on the production of interleukin 6 (IL6) by cultured human endometrial epithelial cells prepared from endometrial biopsy material obtained at different stages in the menstrual cycle was investigated. Basal IL6 production by cells prepared from proliferative endometrium was greater than that produced by cells prepared from secretory endometrium (7.3 +/- 0.3 and 1.1 +/- 0.2 ng/well/24 h respectively, P < 0.001). IL1 (0.025-2.5 ng/ml) caused a dose-dependent increase in IL6 production by cells prepared from both proliferative and secretory endometrium, but cells prepared from secretory endometrium responded to a lower concentration of IL1 than those prepared from proliferative endometrium. IL-1-stimulated IL6 production by epithelial cells prepared from secretory endometrium typically reached 10 times basal values, while in cells prepared from proliferative endometrium stimulated levels were approximately twice the basal values. PP14 (1-50 micrograms/ml) also caused a dose-dependent increase in IL6 production by epithelial cells prepared from secretory endometrium, but had no effect on IL6 production by cells prepared from proliferative endometrium. Even in secretory cells PP14 was less effective than IL1 at stimulating IL6 production, with stimulated levels only reaching twice the basal values. This suggests that PP14 and IL1 act via different mechanisms in the stimulation of IL6 production. The results show that IL6 production by human endometrial epithelial cells is stimulated by other immunomodulatory peptides and this may be part of the network of such peptides in the endometrium which may influence embryo implantation.

Adult↗
PubMed1989

Characterization of an insulin-like growth factor binding protein, analogous to human pregnancy-associated secreted endometrial alpha 1-globulin, in decidua of the baboon (Papio anubis) placenta.

The major secreted protein of the human decidua (pregnancy-associated endometrial alpha 1-globulin [alpha 1-PEG]), is an insulin-like growth factor-binding protein (IGF-BP) that is immunologically and biochemically similar to placental protein 12 (PP12) extracted from term human placenta. Since previous studies have demonstrated that the baboon and human endometrium synthesize and release a number of biochemically and immunologically related polypeptides in culture, this study was undertaken to further characterize a related IGF-BP in baboon placental tissues. Decidua, chorio-amniotic membranes with adhering decidua (CAM-D), and placental villi were obtained from pregnant baboons between Days 134 and 160 of gestation by Cesarean sections. Portions of tissue were either cultured in the presence of 35S-methionine, fixed for immunocytochemistry, or frozen in liquid nitrogen for cytosol extraction. Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) of tissue culture media (TCM) revealed that the major secretory product of the decidua and CAM-D was an acidic polypeptide (Mr 33,000). Western blot analysis and immunoprecipitation of TCM with murine monoclonal antibody (B2H10) against human alpha 1-PEG demonstrated that this molecule, secreted by the baboon decidua and CAM-D, but not the placental villi, was immunologically identical to the human IGF-BP. Immunocytochemical localization of IGF-BP was intense in the cytoplasm of stromal cells in decidua and CAM-D and absent in the placenta. Gel filtration of TCM and cytosol followed by screening of eluates for 125I-IGF-I binding resolved two peaks (Mr greater than 100,000 and 35,000) of specific IGF-BP in decidua and CAM-D. The 35,000 peak had 100-200 times the binding capacity of the Mr greater than 100,000 peak and a Kd of 1.14-1.83 nM. The eluates contained in the Mr 35,000 peak were also immunoreactive to alpha 1-PEG, as accessed by a polyclonal radioimmunoassay. Affinity cross-linking with 125I-IGF-I followed by sodium dodecyl sulfate-PAGE revealed an immunoreactive complex of Mr 36,000, confirming that the baboon protein represents a high affinity IGF-BP. These studies indicate that the hypertrophied stromal cells of the baboon decidua and CAM-D synthesize and release an IGF-BP as their major secretory product, analogous to the situation in humans. The results of this study suggest that this protein may play a role in the regulation of IGF action during pregnancy.

Animals↗
PubMed1995

Fibroblast cells from term human decidua closely resemble endometrial stromal cells: induction of prolactin and insulin-like growth factor binding protein-1 expression.

Term decidual tissue consists of terminally differentiated decidual cells, bone marrow-derived cells, and fibroblast cells. Since undifferentiated decidual cells are fibroblast-like cells of the endometrial stroma, the possibility exists that the fibroblast cells in term decidua are undifferentiated decidual cells. To test this hypothesis, a purified population of fibroblast cells was isolated from term decidua and treated under conditions that are known to induce differentiation of endometrial stromal cells. By flow cytometry and immunocytochemistry, the fibroblast cells from term decidua were shown to be free of cells expressing bone marrow-derived cell-surface antigens and the epithelial cell marker cytokeratin. In addition, they tested positive for the cytoskeletal protein vimentin, thus establishing that they were mesenchymal cells. As with endometrial stromal cells, continuous treatment of the decidual fibroblast cells with the progesterone analog medroxyprogesterone acetate and estradiol in combination with either dibutyryl-cAMP or prostaglandin E2 induced cell aggregation and the expression of prolactin (PRL) and insulin-like growth factor-binding protein-1 (IGFBP-1). When cells were plated at an initial cell density of 0.25 x 10(6) cells/well in a 24-well culture dish with medium changes every three days, PRL was first detected on Days 4-6, and the peak of averaged 24 h-PRL release (30 ng/well) occurred on Days 26-28. The mRNA for decidual fibroblast PRL followed a temporal pattern corresponding to that of the released hormone. The size of the PRL mRNA was 1.15 kb, corresponding to the alternately spliced PRL mRNA reported for decidualized endometrial stromal cells and other extrapituitary sources of PRL.(ABSTRACT TRUNCATED AT 250 WORDS)

Antigens, Surface↗
PubMedDate not supplied

Involvement of selectin-like carbohydrate binding specificity in human gamete interaction.

The recognition of carbohydrate epitopes by complimentary protein receptors has been shown to be a critical factor in gamete interaction in many different animal species. In this study it was hypothesized that, in the human, gamete binding requires an interaction between selectin ligands on the zona pellucida and putative egg binding proteins on the sperm surface. The hemizona assay (a unique internally controlled bioassay that evaluates tight binding of sperm to the zona) and advanced methods of carbohydrate analysis were used to test this hypothesis. From these tests it was shown that oligosaccharide recognition is also required for initial human gamete binding. This study suggests the existence of distinct egg binding proteins on human sperm that can bind to selectin ligands. Additionally, the results suggest a possible convergence in the types of carbohydrate sequences recognized during initial human gamete binding and immune/inflammatory cell interactions. Glycoconjugates that manifest selectin-ligand activity and that express specific carbohydrate epitopes have potent contraceptive and immunosuppressive effects. Such specific oligosaccharide sequences may provide an appropriate recognition signal for embryo development and protection.

Carbohydrate Metabolism↗
PubMed2000

Clinical applications of serum placental protein 14 (PP14) measurement in the IVF-ET cycle.

OBJECTIVE: Placental protein 14 (PP14) is known to be one of the endometrial proteins that reflect endometrial functioning throughout the menstrual cycle. In this study, we examined PP14 as a marker for human endometrial receptivity in order to predict the outcome of in vitro fertilization and the embryo-transfer (IVF-ET) cycle. PATIENTS AND METHODS: The subjects were 72 women who had 96 IVF-ET cycles and who were examined at Tokyo Medical University Hospital during the period of January 1998 to June 1998 because of mechanical or unexplained infertility for a duration of at least 2 years. Serum samples were collected from all patients during treatment cycles, and serum PP14 concentrations were measured by a newly established enzyme-linked immunosorbent assay (ELISA). RESULTS: In the pregnant group, serum PP14 concentrations were markedly increased after ET, and a significant difference between the pregnant group and the nonpregnant group was observed 8 days following ET (p < 0.01). PP14 concentrations were higher in patients with endometria that exhibited homogenous patterns and that were more than 7 mm thicker than in other patients, as determined by ultrasound on the day of oocyte collection (p < 0.005). The pregnancy rates of patients with homogeneous patterns were lower than those of patients showing a trilaminar pattern. No pregnancies were observed when serum PP14 concentrations were greater than 6.85 U/l on the day of oocyte collection. CONCLUSION: PP14 might be a useful marker for human endometrial receptivity to predict the outcome of IVF-ET cycles.

Adult↗
PubMed1985

Distribution of placental protein 14 in tissues and body fluids during pregnancy.

Placental protein 14 (PP14) levels were measured in serum samples from non-pregnant and pregnant women, amniotic fluid, cord blood, and extracts of placenta, decidua and fetal membranes. The levels were low (15-40 micrograms/l) in serum of non-pregnant women. In four pregnancies following in-vitro fertilization, the serum PP14 levels started to rise 2-12 days after embryo replacement. In normal pregnancy, the highest serum PP14 concentrations (up to 2200 micrograms/l) were detected between 6 and 12 weeks. After 16 weeks the level decreased and plateaued at 24 weeks to around 200 micrograms/l. In amniotic fluid, the highest PP14 levels (232 mg/l) were found between 12 and 20 weeks, being considerably higher than those in maternal serum throughout pregnancy. In cord blood, the levels were low (15-22 micrograms/l) or undetectable. In early pregnancy decidua, the PP14 content was higher (41-160 mg/g total protein) than in late pregnancy decidua (60-2700 micrograms/g total protein). In amnion and chorion laeve, the PP14 concentration varied from 50 to 750 and 50 to 1000 micrograms/g protein, respectively. Early pregnancy placenta contained 0.25-15 mg/g and late pregnancy placenta 3-430 micrograms/g protein of PP14. These results show that the levels of PP14 in pregnancy serum have a similar profile to hCG, but in contrast to other placental proteins, the amniotic fluid PP14 levels are remarkably high. This may be explained by suggesting that decidua is a source of PP14.

Amniotic Fluid↗
PubMed1986

Amniotic fluid concentrations of secreted pregnancy-associated endometrial alpha 1- and alpha 2-globulins (alpha 1- and alpha 2-PEG).

The levels of pregnancy-associated endometrial alpha 1- and alpha 2-globulins (alpha 1- and alpha 2-PEG), the two major proteins synthesized and secreted by the endometrium in vitro, have been assayed in 210 amniotic fluid specimens obtained at termination of pregnancy or by amniocentesis, or at delivery. alpha 1-PEG was undetectable until week 10 and thereafter rose to peak levels between weeks 20 and 24. Levels fell 15-fold by week 35 but substantial amounts were still present at parturition. alpha 2-PEG was present at highest levels during early pregnancy, at weeks 6-15, but thereafter levels rapidly fell until during weeks 31-42 alpha 2-PEG was detectable in only 3 of 25 specimens. During weeks 15-20, when alpha 2-PEG levels fell and alpha 1-PEG levels rose, a high correlation was observed between the week of gestation and the log of the ratios of the concentration of these proteins. These observations provide the opportunity to assess the role of endometrial and decidual dysfunction in the aetiology of pregnancy disorders.

Amniotic Fluid↗
PubMed1988

Immunohistological localization of pregnancy-associated endometrial alpha 2-globulin (alpha 2-PEG) in endometrial adenocarcinoma and effect of medroxyprogesterone acetate.

Endometrium from postmenopausal women with endometrial adenocarcinoma was examined immunohistochemically using a monoclonal antibody to pregnancy-associated endometrial alpha 2-globulin (alpha 2-PEG), the major secretory protein of the glandular epithelium during the late luteal phase of the menstrual cycle and early pregnancy. Specimens were obtained at initial diagnostic curettage and at hysterectomy after medroxyprogesterone acetate (MPA) therapy. alpha 2-PEG was not detected in any malignant tissue irrespective of histological differentiation. Non-malignant endometrium obtained in association with malignant tissue was negative for alpha 2-PEG before treatment although after MPA therapy all specimens obtained exhibited marked alpha 2-PEG localization in glands. In four specimens endogenous alkaline phosphatase was observed consistently only in the malignant endometrium. Malignant endometrium does not appear to synthesize alpha 2-PEG nor is its synthesis induced by an oral progestogen, so that it does not represent a useful marker for endometrial carcinoma. Non-malignant endometrium in postmenopausal women appears to be fully capable of alpha 2-PEG production after stimulation with an oral progestogen.

Adenocarcinoma↗
PubMed1993

Concentration of endometrial protein PP14 in uterine flushings throughout the menstrual cycle in normal, fertile women.

OBJECTIVE: To determine the variation in concentration of endometrial protein PP14 in uterine flushings throughout the menstrual cycle comparing this to concentrations in plasma samples. DESIGN: Precise timing of all samples by the luteinising hormone surge. SETTING: Jessop Hospital for Women, Sheffield. SUBJECTS: Twenty-three regularly cycling, previously fertile volunteer women. INTERVENTIONS: Observational study; 10 ml of physiological saline was used to flush the uterine cavity once or serially in the cycle of the study. MAIN OUTCOME MEASURES: The measurement of PP14 levels by radioimmunoassay in uterine flushings and plasma samples. RESULTS: In uterine flushing, PP14 levels were not detectable in significant amounts in the proliferative phase and the early luteal phase; after day LH + 6, the concentration rises rapidly with a doubling time of only 6.6 to 14.6 h in the midluteal phase. In the late luteal phase, the concentrations in uterine flushing were over a hundred times higher than the corresponding plasma samples. CONCLUSIONS: The measurement of PP14 in uterine flushings is likely to be of greater value than the measurement in plasma samples; it may provide a valuable alternative to the evaluation of endometrial function.

Adult↗
PubMed1994

Circulating levels of placental protein 14 in ectopic pregnancy.

OBJECTIVE: To determine circulating levels of placental protein 14 (PP14) in complications of early pregnancy. DESIGN: Biochemical monitoring of women presenting with vaginal bleeding and/or abdominal pain. SETTING: An emergency gynaecological ultrasound clinic in a London teaching hospital. SUBJECTS: Venous blood samples were obtained from 67 women with normal pregnancy (n = 9), ectopic pregnancy (n = 26) and failed intrauterine pregnancy (n = 32). This group included anembryonic pregnancy (n = 18), missed miscarriage (n = 2), spontaneous miscarriage of a previously demonstrated live fetus (n = 6), incomplete miscarriage (n = 4), complete miscarriage (n = 1) and molar pregnancy (n = 1). MAIN OUTCOME MEASURES: Serum PP14 levels in the group of women with a failed intrauterine pregnancy in relation to the normal range for PP14. RESULTS: Eighty-one percent of women who miscarried spontaneously had normal serum PP14 levels; 81% of women with ectopic pregnancy had depressed (< 5th centile) serum PP14 levels. CONCLUSION: Measurements of PP14 may be useful in distinguishing spontaneous miscarriage from ectopic pregnancy, but not in the management of threatened miscarriage.

Abdominal Pain↗
PubMed1996

Serum levels of endometrial proteins during transcervical resection of the endometrium.

OBJECTIVE: To study the potential for dissemination of endometrial tissue substances during transcervical resection of the endometrium (TCRE). DESIGN: Prospective study. SETTING: One university and two county hospitals. PARTICIPANTS: Forty-eight women with dysfunctional bleeding. INTERVENTIONS: The serum levels of two endometrial proteins, insulin-like growth factor binding protein-1 (IGFBP-1) and placental protein 14 (PP14), were measured before and every 10 min during the operations. Blood loss was also measured by a photometer together with absorption of the irrigating fluid containing glycine 1.5% and ethanol 1% by expired-breath tests, and serum sodium and volumetric fluid balances. MAIN OUTCOME MEASURES: Linear correlations between changes in IGFBP-1 and PP14 during TCRE and operating parameters such as operating time, blood loss and fluid absorption. RESULTS: The baseline levels of IGFBP-1 were normal but PP14 could only be detected in one third of the patients, which was due, in part, to pre-operative treatment with danazol. The highest levels of IGFBP-1 and PP14 during surgery correlated positively with the baseline concentrations. Fluid absorption (median 405 ml, range 0-2177) was the only surgical factor associated with increasing serum levels of endometrial proteins. CONCLUSION: Absorption of the solution used to irrigate the uterus is associated with dissemination of endometrial products in the bloodstream during TCRE.

Adult↗
PubMed1993

Diagnosis and management of extrauterine pregnancies.

This study was based on 16 women provisionally diagnosed as having extrauterine pregnancies. Of these, 13 (81.3%) were confirmed as positive at operation. Patients were managed according to 1 of 3 regimens; 1) methotrexate (n = 4), 2) methotrexate followed by surgery (n = 3) and 3) surgery (n = 6). Serial blood samples, collected before and after treatment, were analyzed for ovarian (oestradiol, E2; progesterone, P4) uterine (placental protein 14, PP14) and placental markers (chorionic gonadotrophin, HCG; pregnancy-associated plasma protein-A (PAPP-A). Of the pretreatment samples, only 30.4% and 41.7% were depressed for PP14 and HCG, respectively. By contrast, the diagnostic value of PAPP-A (77.8%) and P4 (87.5%) was greater. Biochemical monitoring of treatment was best achieved with trophoblastic derived antigens (HCG), whereas antigens of maternal origin demonstrated widely varied responses. This study established the effectiveness of chemotherapy for treatment of tubal pregnancies as an alternative to surgery, but if a biochemical marker is required, the marker of choice is HCG.

Abortion, Therapeutic↗