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PubMed1996

Serum relaxin and the major endometrial secretory proteins in in-vitro fertilization and down-regulated hormone-supported and natural cycle frozen embryo transfer.

Relaxin has been postulated to be a modulator of the expression of the endometrial secretory proteins, insulin-like growth factor binding protein (IGFBP-1) and placental protein 14 (PP14). This study evaluated the expression of relaxin in relation to concentrations of these secretory proteins along with oestradiol, progesterone and human chorionic gonadotrophin in groups of pregnant and non-pregnant patients who underwent differing assisted conception treatments. Serum samples were taken from 88 patients at 8 and 12 days after embryo transfer. At 12 days after embryo transfer, relaxin concentrations in the pregnant patients who had undergone in-vitro fertilization (IVF) or natural cycle frozen embryo transfer were significantly higher than those who did not conceive in these groups (mean concentrations 8334 versus 28 and 2608 versus 62 pg/ml respectively, P<0.001). However concentrations in the pregnant patients who had hormone support and transfer of frozen embryos were not significantly different from the patients who did not conceive after the same treatment. Although relaxin expression was associated with corpus luteum activity, it was not related to the number of corpora lutea in IVF patients. A wide range of relaxin concentrations was seen to be compatible with a healthy pregnancy. These serum relaxin concentrations were not found to be directly related to the serum concentrations of IGFBP-1, PP14 or the other factors assessed in this study.

Chorionic Gonadotropin↗
PubMed1996

Insulin-like growth factor-I and alpha 2-macroglobulin in seminal plasma correlate with semen quality.

Insulin-like growth factor-I (IGF-I) and alpha 2-macroglobulin (alpha 2-M) are believed to be involved in the development of germ cells. IGF-I is mainly controlled by concentrations of human growth hormone (HGH), influences cell proliferation and differentiation and its action is mediated by insulin-like growth factor-binding proteins (IGFBP), placental protein 14 (PP14) and prostate-specific antigen (PSA). alpha 2-M acts as a broad spectrum proteinase inhibitor and a binding protein for many cytokines and hormones, e.g. inhibin and activin. This study was designed to identify concentrations of these molecules in seminal plasma in normal semen samples of healthy men, correlations with semen quality, the relationship of IGF-I and alpha 2-M with factors affecting male fertility, and whether vasectomy influences the concentrations of these molecules. Concentrations of IGF-I and alpha 2-M in human seminal plasma were related to semen quality, basal concentrations of HGH, testosterone, IGFBP-3, soluble fibronectin receptor (sFNR), PSA and PP14 in seminal plasma and to serum concentrations of luteinizing hormone (LH) and follicle stimulating hormone (FSH). Commercially available assays were used to analyse 69 semen samples of various quality and 11 post-vasectomy samples. IGF-I concentrations in seminal plasma were significantly correlated with the percentage of morphologically normal spermatozoa (r = 0.748, P = 0.00001) and sperm concentration (r = 0.301, P = 0.011), but negatively correlated with serum FSH (r = -0.506, P = 0.00006) and PSA in seminal plasma (r = -0.388, P = 0.00009). Total alpha 2-M was significantly correlated with sperm count (r = 0.423, P = 0.0005), percentage of progressively motile spermatozoa (r = 0.444, P = 0.00019), quality of motility (sperm motile efficiency, r = 802, P = 0.00001) and straight line velocity (r = 0.411, P = 0.0013). Correlation between the sperm concentration and HGH in seminal plasma was weak (r = 0.287, P = 0.015). Vasectomy reduced the concentration of total alpha 2-M (P = 0.00008) and HGH (P = 0.0068) in the seminal plasma; IGF-I was also reduced after vasectomy when the total ejaculate amount was considered. Thus IGF-I and alpha 2-M are significant for the germ cell development: IGF-I in the maturation of spermatozoa and alpha 2-M in progressive motility.

Follicle Stimulating Hormone↗
PubMed1996

Mechanism of action of the intrauterine contraceptive device: evidence for a specific biochemical deficiency in the endometrium.

The precise mechanism of action of the intrauterine contraceptive device (IUCD) is uncertain. In this study we compared the circulating concentrations of a specific endometrial protein, placental protein 14 (PP14), in 62 women with an IUCD and 16 controls. The concentrations of PP14 were substantially lower in IUCD users. There was no difference in the concentrations of another and less specific endometrial protein, insulin-like growth factor binding protein-1 (IGFBP-1). There was no difference in PP14 concentrations between those women with and without intermenstrual bleeding. We conclude that the reduced concentrations of PP14 in IUCD users reflect defective endometrial function in these women, probably related to the contraceptive effect. We propose that the measurement of PP14 might be a means of comparing the efficiency of different devices.

Adult↗
PubMed1996

The production of tumour necrosis factor alpha (TNF-alpha) by human endometrial cells in culture.

The production of tumour necrosis factor alpha (TNF-alpha) by cultured human endometrial epithelial and stromal cells prepared from endometrium obtained at different stages in the menstrual cycle has been investigated. TNF-alpha was not detectable in the supernatants of stromal cell cultures prepared from endometrial tissue obtained at any time in the menstrual cycle. TNF-alpha production by endometrial epithelial cells in culture varied depending on the time in the cycle at which the endometrial tissue was taken. Cells prepared from tissue obtained during the late proliferative phase of the cycle produced more TNF-alpha than those prepared from tissue obtained at other times in the cycle. In addition, a small increase in TNF-alpha production was seen by cells prepared from tissue obtained during the mid-secretory phase of the cycle. Interleukin 1 (IL-1) (1.4-140 pmol/l) caused a dose-dependent increase in TNF-alpha production by cells prepared from both proliferative and secretory endometrium. Maximum IL-1-stimulated increases in TNF-alpha production were similar in cells from both proliferative and secretory endometrium and typically reached from four to 10 times basal values. High doses of progesterone, either alone or in the presence of oestradiol, also affected TNF-alpha production by epithelial cells. TNF-alpha production by cells prepared from proliferative endometrium was increased by progesterone. In contrast, TNF-alpha production by cells prepared from secretory endometrium was decreased in the presence of progesterone. The effects of steroids on TNF-alpha production were less marked than that of IL-1, with values increasing or decreasing to a maximum of three times the basal value. Placental protein 14 (PP14) (0.18 and 1.8 nmol/l) also increased TNF-alpha production by cells prepared from proliferative tissue, but had no effect on its production by cells prepared from secretory endometrium. PP14-stimulated TNF-alpha levels typically only reached a maximum of two times basal values.

Cells, Cultured↗
PubMed1995

The measurement of CA 125 and placental protein 14 in uterine flushings in women with recurrent miscarriage; relation to endometrial morphology.

The concentrations of CA 125 and placental protein 14 (PP14) were measured in uterine flushings obtained throughout the luteal phase of the cycle from eight normal fertile women. The concentrations of both proteins increased in a similar pattern throughout the luteal phase of the cycle, with the most dramatic increase occurring 6 days after their luteinizing hormone surge (day LH +6). However, a greater variation in CA 125 concentrations was seen compared to that seen for PP14. The concentrations were compared to those obtained on day LH +7 of the cycle from a group (n = 35) of women with recurrent miscarriage. The ranges in concentration of PP14 and CA 125 in the flushings of fertile and recurrent miscarriage patients were very similar. However, a greater proportion of women with recurrent miscarriage (55%) had low concentrations (< 5 ng/ml) of PP14 than in the control group (12.5%) and the concentrations of PP14 in the uterine flushings were significantly less (P < 0.05) in women with recurrent miscarriage compared to the normal fertile group. There was no significant difference in the concentration of CA 125 in the uterine flushings between the two groups. Histological observation of the endometrial biopsy samples from recurrent miscarriage patients gave menstrual cycle datings that ranged from day LH +2.5 to LH +6.5 with retarded endometrium (< day LH +5) in 12 of 35 (34%) patients. Of these 12 patients, 10 (83%) had low PP14 concentrations and six (50%) had low CA 125 concentrations in their uterine flushings. In the recurrent miscarriage patients with histologically normal (> or = day LH +5) endometrial development, 10 out of 23 (43%) also had low PP14 concentrations and 8 out of 23 (35%) had low CA 125 in their uterine flushings. The results suggest that PP14 is better than CA 125 as a marker for endometrial function in this group of women. In some cases (52%) the low concentrations of PP14 in the uterine flushings could be explained by retarded endometrial development but for the others the reduction in PP14 concentration in the uterine flushing was not associated with retardation of endometrial development.

Abortion, Habitual↗
PubMed1995

Relationships between the uterine environment and maternal plasma concentrations of insulin-like growth factor binding protein-1 and placental protein 14 in early pregnancy.

Blood was obtained from 218 women between 6 and 13 weeks of gestation. Measurements of serum insulin-like growth factor binding protein-1 (IGFBP-1) and placental protein 14 (PP14) concentrations were compared with maternal weight and height, maternal smoking habit, indices of maternal haematological status and two placental hormones [human chorionic gonadotrophin (HCG) and human placental lactogen (HPL)]. IGFBP-1 concentration was negatively correlated with maternal weight (P < 0.001) and body mass index (P < 0.001); PP14 concentration was not correlated with these measurements. PP14 concentration was negatively correlated with maternal haemoglobin concentration (P = 0.010), mean corpuscular volume (P = 0.003) and serum ferritin concentration (P = 0.016). The concentrations of PP14 were significantly less among smokers (P < 0.001); IGFBP-1 concentrations were uninfluenced by smoking. IGFBP-1 concentration was positively correlated with maternal serum HCG (P = 0.003) and maternal serum HPL (P = 0.002). PP14 concentration was positively correlated with maternal serum HCG (P < 0.0001) but not with HPL. These findings demonstrate that the maternal environment has an early influence on both endometrial and placental function.

Body Mass Index↗
PubMed1995

Variations in concentrations of the major endometrial secretory proteins (placental protein 14 and insulin-like growth factor binding protein-1) in assisted conception regimes.

We have previously shown that placental protein 14 (PP14) concentrations were depressed in two pregnancies that followed down-regulation of the anterior pituitary and exogenous hormone support prior to a frozen-thawed embryo transfer. We now report on a more comprehensive series of pregnancies following this form of treatment, in-vitro fertilization (IVF) and natural cycle frozen-thawed embryo transfer. Serum specimens were analysed for PP14 and insulin-like growth factor binding protein-1 12 days after embryo transfer and at 7 weeks gestation. At 12 days after embryo transfer, the mean serum PP14 concentrations in the IVF and natural cycle were significantly higher in those who conceived than those who did not (82 versus 23 and 107 versus 39 micrograms/l respectively, P < 0.001). Although the mean PP14 concentration in the hormone-supported pregnant patients was higher than in the non-pregnant patients, this had not reached statistical significance 12 days after embryo transfer (49 versus 31 micrograms/l). By 7 weeks gestation the PP14 concentrations in the hormone-supported pregnant patients were significantly higher than in the non-pregnant patients (152 versus 31 micrograms/l, P < 0.001). However, the PP14 concentrations for hormone-supported pregnant patients were significantly lower (P < 0.001) than those for pregnant IVF or natural cycle patients at 7 weeks gestation (152, 777 and 660 micrograms/l respectively). The PP14 concentrations in the pregnant patients, although lower than those in IVF and natural cycle pregnancies, were higher than those previously reported in ovarian failure and Turner's syndrome ovum donation cycles.(ABSTRACT TRUNCATED AT 250 WORDS)

Carrier Proteins↗
PubMed1995

Endometrial and placental protein markers and ovarian steroids in serum during in-vitro fertilization cycles.

The objective of this study was to find the earliest time at which it was possible to detect clinical pregnancy in an in-vitro fertilization (IVF) treatment cycle supported with human chorionic gonadotrophin (HCG), and also retrospectively to diagnose abnormal ovarian- or endometrium-related situations in failure cycles. Serum samples were taken in 41 IVF cycles at frequent intervals from the beginning of ovarian stimulation until menstrual bleeding occurred or a pregnancy was established. Concentrations of oestradiol, progesterone, placental protein 14 (PP14), pregnancy-specific beta 1-glycoprotein (SP1), and pregnancy-associated plasma protein A (PAPP-A) were determined in the serum samples using commercially available (steroid) or purpose-developed (protein) immunoassays. The cycles were retrospectively distributed into four outcome groups: (i) fertilization failure (FF, n = 8); (ii) implantation failure (IF, n = 10); (iii) 'interaction' (embryo-endometrium) cycle (IC, n = 14), and (iv) clinical pregnancy (CP, n = 9). The embryo-endometrium interaction was detected by a rise in SP1 in 23 cycles (70% of embryo transfers) at a time when endogenous HCG was still masked by external support. Early ('false') positive SP1 concentrations were observed in two out of eight and five out of 14 cases in groups FF and IC respectively, but never amongst the ongoing pregnancies (CP). PAPP-A did not distinguish pregnancy from the other outcomes. The PP14/progesterone ratio was lower, later in the cycle, in CP than in the other groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1987

Micronized oral progesterone increases the circulating level of endometrial secretory PP14/beta-lactoglobulin homologue.

The effect of luteal phase supplementation of micronized oral progesterone, 200 mg daily, on the serum levels of endometrial secretory placental protein 14 (PP14)/beta-lactoglobulin homologue was studied in a double blind/cross-over fashion on five infertile women with apparently normal ovulatory cycles. Blood samples were taken on cycle days 10-12, 20-22 and 24-27, and the serum progesterone and PP14 concentrations were measured by specific radioimmunoassays. In each progesterone cycle the serum progesterone levels were higher than in the corresponding placebo cycle. In each case, the serum concentration of endometrial PP14 was also higher during progesterone than placebo treatment, but this was seen in the late luteal phase only. In view of the previous demonstration of endometrial synthesis of PP14 and the immunological identity between PP14 and progestogen-dependent endometrial protein, these results indicate that endometrial protein secretion can be increased by micronized oral progesterone in infertile ovulatory women whose serum progesterone level is within the normal range.

Administration, Oral↗
PubMed1987

The effect of Danazol on the circulating levels of 34K insulin-like growth factor-binding protein (PP12) and endometrial secretory protein PP14.

Twenty-four women, 11 with endometriosis and 13 with fibrocystic mastopathy, were treated with a medium dose (300-400 mg daily) of Danazol for 6 months. The circulating level of progesterone, the 34K insulin-like growth factor-binding protein (IGF-bp) and endometrial protein PP14 (placental protein 14) were measured by specific radioimmunoassays before and during treatment. The serum progesterone concentration decreased significantly during Danazol treatment, as did the serum levels of 34K IGF-bp and PP14. By the third month of therapy amenorrhoea was observed in 22 out of 24 women and this was accompanied by a further decline in the IGF-bp and PP14 levels. In light of the previous observations on the IGF-bp and PP14 synthesis by secretory endometrium and the fact that Danazol causes endometrial atrophy, these results suggest that Danazol treatment has an effect on endometrial protein secretion.

Adult↗
PubMed1987

The post-menopausal uterus: the effect of hormone replacement therapy on the serum levels of secretory endometrial protein PP14/beta-lactoglobulin homologue.

The effect of cyclical oestrogen-progestogen replacement on the levels of endometrial secretory protein PP14 (placental protein 14) was studied in 39 post-menopausal women, 13 of whom had had a hysterectomy. In those women with an intact uterus, the average rise of the late luteal phase serum PP14 concentration was from 29 to 45 micrograms/l, a 55% rise (P less than 0.001). Only a slight rise (from 27 to 30 micrograms/l) was observed in hysterectomized women. Cyclical replacement with oestrogen and levonorgestrel caused a greater rise in serum PP14 level than did replacement with the same dose of oestrogen plus medroxyprogesterone acetate (P less than 0.05). Because PP14 is found also in the serum of hysterectomized post-menopausal women, the uterus cannot be the only source of circulating PP14. But, the difference between hysterectomized and non-hysterectomized women indicates that the quiescent post-menopausal uterus responds to oestrogen-progestogen replacement by increased PP14 secretion, which can be detected and quantified by a blood test. The measurement of PP14 in serum may also become useful for testing the effect of various progestogens.

Estradiol↗
PubMed1989

Suppression of prolactin secretion during ovarian hyperstimulation is followed by elevated serum levels of endometrial protein PP14 in the late luteal phase.

A double-blind placebo-controlled study on bromocriptine administration during days 2-12 of ovarian hyperstimulation for in-vitro fertilization (IVF) showed that, in bromocriptine cycles, levels of the endometrial protein PP14 were higher in the late luteal phase. This was verified both by calculating forward from the day of human chorionic gonadotrophin (HCG) administration and backward from the onset of the next period. Bromocriptine had no effect on IVF performance. During bromocriptine treatment the serum prolactin levels declined and serum oestradiol levels were higher on day 9 of the cycle. There was a positive correlation (r = 0.55; P = 0.012) between the serum oestradiol levels on day 9 and the PP14 levels on days 22-23 of the cycle. No difference was found in the luteal phase progesterone levels between bromocriptine- and placebo-treated cycles. These results suggest that low prolactin and/or high oestradiol levels during the follicular phase have an influence on the subsequent secretory capacity of the endometrium as reflected by secretion of a specific endometrial protein.

Bromocriptine↗
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 non-reproduction-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

Endometrial assessment in a group of infertile women on stimulated cycles for IVF: immunohistochemical findings.

An immunohistochemical assessment of the endometrium was carried out in a group of IVF patients on stimulated cycles, in order to evaluate this technique against standard histological methods and to consider its application in a clinical situation. Monoclonal antibodies to the two cycle-dependent proteins: pregnancy associated endometrial alpha 2-globulin (alpha 2-PEG) and 24K (a protein originally isolated from an oestrogen-dependent breast tumour line, MCF-7) were used in the experiment. Immunohistochemical results concerning the effect of drug stimulation, age and date of biopsy on the secretory state of the endometrium revealed trends which were consistent with previous histological data, helping to confirm the value of this new technique. In addition, several specimens were found to have a normal, i.e. in phase, histological appearance but to have an atypical pattern of protein secretion. These observations suggest that biochemical monitoring of the uterus should be used in conjunction with routine histological dating.

Adult↗
PubMed1990

Serum PP14 levels in a patient with Turner's syndrome pregnant after frozen embryo transfer.

Data are presented on serum levels of the pregnancy-associated placental protein (PP14) in an XO mother, pregnant after frozen embryo transfer. Steroid replacement before and during early pregnancy was provided with transdermal oestradiol and vaginal micronized progesterone. Characterization of a pre-treatment cycle indicated a physiological response by the endometrium. The pregnancy was associated with normal maternal blood levels of all hormones considered responsible for maintenance of early gestation but with consistently subnormal serum levels of PP14. Hitherto, PP14 has been believed to be an endometrial protein arising from the decidua during pregnancy, under the influence of ovarian steroids. The findings in this case suggest that other factors are involved in its production, possibly under the control of, or originating from, the maternal ovary.

Adult↗
PubMed1990

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.

Adult↗
PubMed1990

Serum levels of pregnancy-associated endometrial alpha 2-globulin (alpha 2-PEG), a glycosylated beta-lactoglobulin homologue, in successful and unsuccessful assisted conception.

Pregnancy-associated endometrial alpha 2-globulin (alpha 2-PEG) is the major secretory protein of the endometrium in the late luteal phase. Serum levels were measured during the implantation period in successful and unsuccessful cycles of assisted conception. The effects of two different treatment regimens were also compared. All patients who underwent ovulation induction therapy showed an endometrial response as measured by a rise in serum levels of alpha 2-PEG. This rise was the same regardless of either implantation or the method of ovulation induction used. Higher levels were seen at oocyte recovery in those cycles treated with clomiphene citrate and may reflect a disordered endometrial response involving a direct effect of the drug on the endometrium. This may explain the difference in success rates in assisted conception when an alternative treatment initially involving down-regulation of endogenous gonadotrophin activity and subsequent administration of gonadotrophins is used. The rise in serum levels seen in pregnancy did not differ statistically according to whether the pregnancy progressed normally or subsequently aborted. Considerably reduced alpha 2-PEG production by the endometrium was identified in one patient who subsequently aborted. The measurement of serum levels of alpha 2-PEG during the implantation phase is unlikely to provide a clinical tool either for early detection of pregnancy or as a prognostic factor of successful implantation.

Abortion, Spontaneous↗