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PubMed1990

Immunohistological distribution of the secretory endometrial protein, 'pregnancy-associated endometrial alpha 2-globulin', a glycosylated beta-lactoglobulin homologue, in the human fetus and adult employing monoclonal antibodies.

We have previously demonstrated that pregnancy-associated endometrial alpha 2-globulin (alpha 2-PEG), the human glycosylated beta-lactoglobulin homologue (HG-BLG), is quantitatively the major secretory soluble protein product of the secretory endometrium during the latter half of the menstrual cycle and decidua spongiosa of the gestational endometrium during early pregnancy, and is principally localized to the glandular epithelium. In the present study employing monoclonal antibodies in immunohistological techniques, the distribution and localization has been examined in normal and pathological tissues of the adult and first-trimester fetus. No significant staining for alpha 2-PEG was detected in any nonreproduction-associated tissue in the normal adult nor any tissue in the fetus. In the adult, most intense staining was associated with the endometrial glandular epithelium in the uterus or in ectopic sites in patients with endometriosis. During the menstrual cycle and pregnancy, appearance of alpha 2-PEG in endometriosis was strongly linked with its appearance in uterine endometrial tissue, suggesting that endometriotic tissue exhibited competence to respond to the same hormonal milieu required to induce synthesis in the uterine endometrium. Localization to the mucosal epithelium of the Fallopian tube was consistent with synthesis of alpha 2-PEG, albeit at low levels, and staining at this site reflected fluctuations of staining within the uterus. Of the pathological specimens examined, staining was only detected in a proportion of ovarian carcinomas. No staining was detected in the mammary gland, a site of beta-lactoglobulin synthesis, whether obtained during pregnancy or lactation.(ABSTRACT TRUNCATED AT 250 WORDS)

Antibodies, Monoclonal↗
PubMed1990

Binding proteins for insulin-like growth factors in the human ovary: identification, follicular fluid levels and immunohistological localization of the 29-32 kd type 1 binding protein, IGF-bp1.

The occurrence of pregnancy-associated endometrial alpha 1-globulin (alpha 1-PEG), a 29-32 kd insulin-like growth factor binding protein, now termed type 1 or IGF-bp1, has been examined in the human ovary by monoclonal and polyclonal antibody based radioimmunoassay and immunohistological techniques. Follicular fluids aspirated from 51 follicles of 32 women undergoing hyperstimulation involving buserelin or clomiphene-based protocols contained 35.5-276.0 ng/ml (mean 101.0 mg/ml) of immunoreactive IGF-bp1. Mean fluid concentrations were three times the level of IGF-bp1 detected in paired serum samples, available for 21 women. Immunoreactive IGF-bp1 in follicular fluid exhibited similar dose-response curves to purified protein and amniotic fluid and immunoreactive IGF-bp1 coeluted in gel filtration with a peak of [125I]-IGF-1 binding corresponding to the elution profile of purified IGF-bp1. Gel filtration also revealed the presence in follicular fluid of a greater than 100 kd binding protein with a binding capacity equal to IGF-bp1 under the conditions employed. A highly significant correlation (P less than 0.001) was found between follicular fluid progesterone and IGF-bp1 and a correlation of lower significance was found between oestradiol and IGF-bp1 (P less than 0.05). However, only low levels of immunoreactive IGF-bp1 were detected in supernatant media of granulosa cells in culture (range undetectable to 2.3 ng/ml). Employing monoclonal antibody-based immunohistology, immunoreactive IGF-bp1 was consistently associated with luteinized granulosa cells of corpora lutea rather than paraluteal cells and its intensity of reactivity appeared to reflect luteal phase steroid hormone profiles. No consistent reactivity was detected in preovulatory follicles and granulosa cells in culture, although reactivity was associated with primordial oocytes. Immunoreactive IGF-bp1 was detected in six of nine supernatant media of explants of luteal tissue obtained from five corpora lutea, with levels ranging from undetectable to greater than 200 ng/ml. These observations suggest that IGF-bp1 is primarily related to luteinization of the granulosa and the resultant luteal cells, and if produced by the luteal cells, additional exogenous factors are required to induce production by granulosa cells in vitro.

Adult↗
PubMed1991

Expression of endometrial protein PP14 in pelvic and ovarian endometriotic implants.

Localization and immunohistochemical staining patterns of endometrial protein PP14 were studied in 55 patients providing 86 histologically confirmed pelvic and ovarian endometriotic implants. The cellular localization of PP14 in endometriotic implants is comparable with that in the endometrium: epithelial cells express PP14, whereas stromal cells are negative. Positive immunostaining is restricted to apical secretory-like granules of the epithelial cells. In endometriotic implants with positive staining, PP14 is expressed by some, but not all, glandular epithelial cells. Furthermore, positive staining for PP14 is observed most frequently in foci with in-situ secretory differentiation, whereas implants with proliferative or atrophic implants only rarely express PP14. Finally, PP14 can be localized in endometriotic foci at different depths of infiltration, although positive labelling is seen more often in superficial implants. These data demonstrate that PP14 can be expressed by endometriotic glandular epithelial cells with secretory cellular differentiation and that the histological appearance of ectopic implants sometimes only poorly reflects their function.

Endometriosis↗
PubMed1991

Evaluation of the luteal phase.

The values of various methods used to evaluate the luteal phase, including basal body temperature, measurement of progesterone (P), endometrial biopsy, ultrasonographic measurement of endometrial thickness, and measurement of endometrial proteins, are reviewed. Luteal phase defect (LPD) is a controversial entity. The diagnosis of this condition is best based on a histological study of the endometrium. Methods to improve the accuracy of the diagnosis are discussed. LPD is more likely to be a result of an abnormal response of the endometrium to P, than to a subnormal production of P by the corpus luteum. Many methods of treatment for LPD have been proposed but none is based on a properly controlled clinical trial. Treatment designed to improve the response of the endometrium to P may be more rewarding than P supplementation.

Biopsy↗
PubMed1991

Correlation of serum progestagen-associated endometrial protein levels with endometrial biopsies serum steroid levels and therapy for luteal phase defects.

The progestagen-associated endometrial protein (PEP) level rises from the early to the late luteal phase. A study was performed in infertile women where late luteal phase endometrial biopsies and serum PEP levels were obtained. The objective of the study was to evaluate the correlation between the PEP levels and the endometrial biopsies and to determine if subnormal PEP levels could be improved by the same therapies used to correct endometrial defects. There was a poor correlation between PEP levels and endometrial biopsies (r = 0.17). Similarly, there was no significant correlation between PEP levels and levels of the following hormones: mid- and late-luteal phase progesterone (P) (r = 0.186 and 0.282 respectively), mid-luteal phase 17-hydroxyprogesterone (17-OHP) (r = 0.139) and mid-luteal phase oestradiol (r = 0.135). Furthermore, there was no correlation between PEP levels and the dosage of progesterone used in therapy (r = 0.07). There were no statistically significant differences in PEP values (U/ml) depending on whether any fertility drug was taken. Thus our data suggest that progesterone may only have a permissive role, with some other factor(s) controlling the actual rise and fall of PEP.

17-alpha-Hydroxyprogesterone↗
PubMed1991

Lack of placental protein 14 production in pregnancy after frozen embryo transfer, down-regulation of anterior pituitary and administration of exogenous oestradiol and progesterone.

The circulating levels of placental protein 14 (PP14) and progesterone were measured in three pregnancies resulting from the transfer of cryopreserved embryos. Two of these women had suppressed ovarian activity as a result of pituitary down-regulation with the luteinizing hormone-releasing hormone agonist (buserelin) prior to treatment with exogenous oestradiol and progesterone. After 14 days of oral oestradiol treatment and if the endometrial thickness was greater than 7 mm, progesterone was given intramuscularly for a further 14 days with embryo transfer on the third day of this treatment. On confirmation of pregnancy by human chorionic gonadotrophin analysis, progesterone administration was altered to transvaginal pessaries for maintenance of adequate progesterone levels and endometrial support. In the two women with ovarian suppression, PP14 levels remained below the 2.5th centile of the normal range for pregnancy. In the third pregnancy, embryo transfer was performed 3 days after a spontaneous luteinizing hormone surge in a normal menstrual cycle. In this pregnancy, PP14 levels were within the normal range. Ultrasonic examination confirmed three normal ongoing singleton pregnancies. These results suggest that the majority of PP14 production in normal pregnancy is under ovarian or anterior pituitary control and that the influence of progesterone is of a secondary nature.

Buserelin↗
PubMed1991

The role of measurement of progestagen-associated endometrial protein in predicting adequate endometrial differentiation.

Late secretory endometrium synthesizes and secretes progestagen-associated endometrial protein (PEP), which is measurable in the serum of cycling women, and has been shown to increase in concentration during the luteal phase of the normal menstrual cycle. The purpose of this study was to determine the utility of serum PEP as a predictor of normal or inadequate luteal phase endometrial differentiation. One-hundred-and-twenty-five endometrial biopsies were taken within 4 days of a subsequent menstrual period, during the course of evaluation of infertility or recurrent abortion. Twenty-one of these biopsies demonstrated glandular/stromal asynchrony. Thirteen patients exhibited 'out-of-phase' endometrial biopsies and 91 patients had 'in-phase' biopsies. Mean PEP values significantly increased from the mid-luteal phase to the late luteal phase. Mean PEP values for patients with in-phase biopsies (43.92 units/ml) were not significantly different from those with out-of-phase biopsies (25.24 units/ml, P = 0.0942). PEP values for patients with asynchronous biopsies were intermediate. Use of clomiphene citrate did not affect these results.

Biopsy↗
PubMed1991

Effect of clomiphene citrate on the synthesis and release of the human beta-lactoglobulin homologue, pregnancy associated endometrial alpha 2-globulin, by the uterine endometrium.

As an extension to a previous study to examine the effects of clomiphene citrate (CC) on human chorionic gonadotrophin/luteinizing hormone (LH) receptors in human corpora lutea, we sought to determine the effect of CC on the synthesis of the beta-lactoglobulin homologue, pregnancy associated endometrial alpha 2-globulin (alpha 2-PEG) by the luteal phase human endometrium. Seventeen normally-cycling women aged 25.9 +/- 2.1 years (mean +/- SE), undergoing elective tubal ligation, were divided into four groups consisting of controls (n = 2) and three treatment groups. Each of the treatment groups (n = 5/group) was treated with either 50 mg (group 1), 100 mg (group 2) or 150 mg (group 3) of CC daily on days 5 to 9 of the cycle. Portions of endometrial tissue were obtained by curettage 7 to 9 days following the urinary LH surge and were either fixed for histological analyses or cultured in vitro in the presence of [35S]methionine. Tissue media (TCM) that were analysed by two-dimensional gel electrophoresis followed by fluorography indicated that the synthesis of alpha 2-PEG was most markedly reduced in groups 2 and 3. Densitometric scans of TCM Western blots and radioimmunoassay (RIA) of TCM showed that the overall synthesis of alpha 2-PEG decreased from 5.8 +/- 1.9 arbitrary units (AU) and 371.2 +/- 239.3 ng/ml in group 1 to 3.1 +/- 0.8 AU and 69.4 +/- 24.9 ng/ml, and 2.7 +/- 1.1 AU and 52.4 +/- 19.8 ng/ml in groups 2 and 3 respectively. On cycle days 20-22, control values were 5.8 AU and 498.1 ng/ml respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1991

Secretory endometrial protein PP14 in women with early pregnancy bleeding.

The decidualized endometrium produces secretory proteins of which secretory endometrial protein PP14 is the major product during the first trimester of pregnancy. The protein is secreted into the uterine lumen as well as into the peripheral blood. The purpose of this study was to examine whether decidual function, evaluated by the serum concentration of PP14, was different in women with early pregnancy bleeding compared to normal pregnant women. A reference range for serum PP14 was established on the basis of single samples from 236 normal pregnant women with ultrasonically confirmed gestational age. All the women were delivered of a normal child at term. The study comprised 128 pregnant women admitted because of vaginal bleeding between 6 and 18 weeks gestation. At ultrasonography, intrauterine fetal heart activity was either present or was confirmed at a subsequent examination. No difference was found in the serum level of PP14 compared to that in normal pregnancies, but women with vaginal bleeding and depressed PP14 levels appeared to have a 5-fold higher risk of preterm delivery than women with bleeding and normal PP14 levels.

Adult↗
PubMed1992

Placental protein 14 in human in-vitro fertilization early pregnancies.

Placental protein 14 (PP14) and human chorionic gonadotrophin (HCG) were analysed in patients participating in an in-vitro fertilization-embryo transfer programme which did not include any kind of luteal support. Women with normal pregnancies, spontaneous abortions, ectopic pregnancies, biochemical pregnancies and non-pregnant women were compared. A combination of HCG and PP14 analyses distinguished between normal and abnormal implantation as early as 15 days after oocyte retrieval. The product of HCG (IU/l) and PP14 (micrograms/l) concentrations differed significantly between normal pregnancy, spontaneous abortion and ectopic pregnancy (P = 0.0248). It is concluded that both endometrial (PP14) and trophoblastic (HCG) markers, when used in combination, exhibit changes in abnormal implantation which may be clinically useful.

Abortion, Spontaneous↗
PubMed1992

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.

Adult↗
PubMed1992

Serum placental protein 14 (PP14) reflects endometrial status during hormone replacement therapy.

Healthy post-menopausal women were randomly assigned to treatment groups receiving 28 day treatment cycles of oestradiol valerate (2 mg, days 1-28) sequentially combined with levonorgestrel (75 micrograms, days 17-28) (n = 19); oestradiol valerate (2 mg, days 1-28) continuously combined with cyproterone acetate (1 mg, days 1-28) (n = 18), or placebo (n = 22). Treatment continued for 2 years. After therapy, the women receiving the sequential combination had an early secretory phase (74%) or atrophic endometrium (26%). All women receiving the continuous combination or placebo had inactive or atrophic endometrium, or the tissue was inadequate for assessment. Biochemical parameters of endometrial secretion [oestradiol dehydrogenase (EDH), isocitrate dehydrogenase (ICDH)] measured in endometrial tissue and placental protein (PP14) measured in serum were all low in the groups receiving the continuous combination and placebo. During sequential treatment, all biochemical parameters showed significantly higher mean values. Serum PP14 accurately reflected this pattern. Furthermore, in the sequentially treated group, serum PP14 correlated highly significantly with EDH (r = 0.70; P less than 0.001) and ICDH (r = 0.57; P = 0.01). These correlations were of the same magnitude as the correlations between the endometrial markers, which indicates that serum PP14 reflects the endometrial status. Furthermore, serum PP14 measurements at 3 months and at 2 years of treatment were highly correlated, reflecting the long-term validity of the measurement. The present data suggest that PP14 measured in serum may be used to assess endometrial status. Further studies are required to determine whether serum PP14 will discriminate pathological endometrial conditions.

Cyproterone Acetate↗
PubMed1993

The correlation of placental protein 14 concentrations in uterine flushing and endometrial morphology in the peri-implantation period.

The relationship between the concentrations of placental protein 14 (PP14) in uterine flushing and the endometrial morphology in the mid-luteal phase was assessed in a prospectively designed study involving the precise timing of all samples by the luteinizing hormone (LH) surge. A total of 29 regularly cycling women with unexplained infertility or recurrent miscarriage were studied. To flush the uterine cavity, 10 ml of physiological saline solution was used immediately prior to sampling of an endometrial specimen for morphological study, in the mid-luteal phase. PP14 concentrations were measured by radioimmunoassay in uterine flushings and plasma samples; the endometrium was assessed by the use of histological dating criteria and morphometric techniques. PP14 levels in uterine flushings were correlated with endometrial dating and volume fraction measurement of the glands. They were consistently below the sensitivity of the assay with histological dating of < day LH +5, or when the glandular lumen occupied < 20% of the gland. In contrast, PP14 concentrations in plasma were not related to histological dating or morphometric analyses, and did not differ in patients with normal endometrial development (20.8 ng/ml) and in those with retarded endometrial development (22.5 ng/ml). The presence of detectable concentrations of PP14 in uterine flushing was significantly associated with normal histological dating. Uterine flushing may therefore provide a reliable, non-invasive alternative to endometrial biopsy in the evaluation of endometrial function in the peri-implantation period.

Adult↗
PubMed1993

Function of the corpus luteum, the endometrium and the trophoblast after treatment of tubal pregnancy by prostaglandin F2 alpha.

The activity of the corpus luteum, the endometrium and the trophoblast was studied after local medical treatment of 31 women with tubal pregnancy. We measured the serum concentration of progesterone, the secretory endometrial protein placental protein 14 (PP14), and human chorionic gonadotrophin (HCG) before and after treatment by injection of prostaglandin F2 alpha into the site of the gestation and into the corpus luteum. There was no significant difference in the pre-treatment serum progesterone and serum PP14 concentrations of 26 women who were treated successfully and of five women, who were operated on after failure of the treatment. After the prostaglandin treatment the serum progesterone and PP14 concentrations decreased simultaneously with the serum HCG concentration or remained at a low, constant concentration. We conclude that measurement of serum progesterone and PP14 cannot be used for selection of patients for treatment by prostaglandin F2 alpha or for monitoring the effect of the treatment. The injection of prostaglandin into the ovary has either no effect on the activity of the corpus luteum or induces only a partial luteolysis.

Adult↗
PubMed1993

Effect of tamoxifen alone and in combination with RU 486 on the endometrium in the mid-luteal phase.

The effects of RU 486 combined with tamoxifen and tamoxifen alone on hormonal parameters and endometrial development at the time of implantation were studied. Measurements of cytosolic oestrogen and progesterone receptors in endometrium and placental protein 14 (PP14) in plasma were also included. Three dosage schedules were used: single oral dose of 40 mg tamoxifen alone and in combination with 200 mg RU 486, and 40 mg tamoxifen for three consecutive days starting on the first day after the luteinizing hormone (LH) surge. The combined treatment prolonged the luteal phase (P < 0.05) and increased the plasma levels of progesterone. A single dose of tamoxifen did not affect the bleeding pattern and plasma hormone levels, but raised plasma oestradiol and LH with the 3-day treatment. The endometrium was retarded after the combined and the 3-day treatment with tamoxifen. Concentrations of cytosolic progesterone receptors were higher after the combined therapy, but were unaffected after tamoxifen only. PP14 levels were higher (P < 0.05) after repeated tamoxifen doses than in controls, but were lower with combined treatment. Progesterone and oestrogen are evidently necessary for endometrial maturation during the secretory phase of the menstrual cycle. PP14 levels in plasma cannot be used for clinical assessments of endometrial function because high levels coincide with disturbed endometrial development.

Embryo Implantation↗
PubMed1993

Circulating placental protein 14: in the first trimester of spontaneous and IVF pregnancies.

Circulating placental protein 14 (PP14) levels were measured during the first trimester in three groups of pregnant women: (i) natural conception (n = 15); (ii) pituitary desensitization with buserelin and ovarian stimulation with human menopausal gonadotrophin (HMG) followed by in-vitro fertilization and embryo transfer (IVF-ET) (n = 15); and (iii) ovarian stimulation with clomiphene citrate and HMG, followed by IVF-ET (n = 16). A 7- to 8-fold increase in serum PP14 levels was observed in normal pregnancies between weeks 4 and 10. This increase was earlier and less marked in group (ii) and absent in group (iii). These findings support the concept that endometrial function is altered in pregnancies achieved following ovarian stimulation. Alternatively, if the ovary is an important source of PP14, then these data suggest that in contrast to ovarian synthesis of steroids and the peptide relaxin, ovarian stimulation results in an impairment of PP14 synthesis, and that this is most marked when clomiphene citrate has been used.

Adult↗
PubMed1993

Serum placental protein 14 concentrations in the first trimester of ovum donation pregnancies.

The concentration of the endometrial glycoprotein placental protein 14 (PP14) has been measured at weekly intervals during the first trimester in serum obtained from women who had become pregnant naturally (n = 15, all singleton) or following ovum donation (n = 16, 15 singleton and one twin) after ovarian failure (n = 13), Turner's syndrome (n = 2) or post-chemotherapy (n = 1). The concentration of PP14 failed to rise in ovum donation pregnancies, and was significantly reduced compared to the concentration in natural conceptions from 5 to 13 weeks gestation (P < 0.05-0.01). These findings demonstrate that the corpus luteum is essential for the normal increase in serum PP14 in the first trimester. Furthermore, they suggest that progesterone is not the major stimulus to PP14 synthesis in early pregnancy.

Female↗