Endometrial protein PP14 and CA-125 in recurrent miscarriage patients; correlation with pregnancy outcome

The concentrations of endometrial proteins PP14 and CA-125 were measured in uterine flushings taken on days LH+10 and LH+12 (10 and 12 days after luteinizing hormone surge) of the menstrual cycle from 15 normal, fertile women and 49 women who suffered recurrent miscarriage. The concentration of PP14 was significantly lower in the flushings from the recurrent miscarriage patients than in those from fertile controls on both day LH+10 (median: 1300, range: 3-10 300 ng/ml versus median: 13 933, range: 2174-40 404 ng/ml; P < 0.01) and LH+12 (median: 1560, range: 820-12 100 ng/ml versus median: 14 047, range 1402-62 108 ng/ml; P < 0.05). Similarly concentrations of CA-125 were significantly lower in flushings from recurrent miscarriage women compared to controls on both day LH + 10 (median: 1555, range: 47-6710 U/ml versus median: 6385.5, range 2884-27 731 U/ml, P < 0.01) and LH+12 (median: 2892, range: 956-9974 U/ml versus median: 7127.5, range: 1591-21 343 U/ml; P < 0.05). In contrast there was no significant difference in the concentration of PP14 in plasma samples taken on the same days as the flushings from recurrent miscarriage patients and fertile controls. The concentrations of PP14 in uterine flushings obtained on day LH + 10 or LH + 12 from recurrent miscarriage women during a pre-pregnancy investigative cycle were significantly lower (P < 0.05) in patients who went on to miscarry (median: 1000, range: 9-2900 ng/ml) than those who went on to have a live birth (median: 1440, range: 4-12 100 ng/ml) during a subsequent pregnancy. In contrast there was no significant difference in uterine CA-125 or plasma PP14 concentrations between these two groups of recurrent miscarriage patients. The results suggest that measurements of uterine PP14 and CA-125 may be useful in the assessment of endometrial development in recurrent miscarriage patients and suggest the importance of PP14 in preparing the endometrium for embryo implantation. In addition pre-pregnancy uterine PP14 measurements may be useful in predicting subsequent pregnancy outcome.      

Implantation: 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 protein14 (PP14) in uterine flushing and the endometrial morphologyin the mid-luteal phase was assessed in a prospectively designedstudy involving the precise timing of all samples by the luteinizinghormone (LH) surge. A total of 29 regularly cycling women withunexplained infertility or recurrent miscarriage were studied.To flush the uterine cavity, 10 ml of physiological saline solutionwas used immediately prior to sampling of an endometrial specimenfor morphological study, in the mid-luteal phase. PP14 concentrationswere measured by radioimmunoassay in uterine flushings and plasmasamples; the endometrium was assessed by the use of histologicaldating criteria and morphometric techniques. PP14 levels inuterine flushings were correlated with endometrial dating andvolume fraction measurement of the glands. They were consistentlybelow the sensitivity of the assay with histological datingof < day LH +5, or when the glandular lumen occupied <20%of the gland. In contrast, PP14 concentrations in plasma werenot 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 PP14in uterine flushing was significantly associated with normalhistological dating. Uterine flushing may therefore providea reliable, non-invasive alternative to endometrial biopsy inthe evaluation of endometrial function in the peri-implantationperiod.     

The production of leukaemia inhibitory factor by human endometrium: presence in uterine flushings and production by cells in culture

The concentration of leukaemia inhibitory factor (LIF) was measured in uterine flushings obtained from normal fertile women, from women with unexplained infertility and from women who suffered recurrent miscarriage. In normal fertile women, LIF was not detected in flushings obtained on days luteinizing hormone (LH)+0 to LH+6 of the cycle, but concentrations gradually increased from day LH+7 to a maximum at day LH+12. The amount of LIF in flushings obtained from women with unexplained infertility was significantly lower than in those from normal fertile women on day LH+10 (P < 0.05). The production of LIF by cultured human epithelial and stromal cells was also investigated. LIF was not detectable in the supernatants of cultured stromal cells. Basal LIF production by epithelial cells varied according to the stage in the cycle at which the biopsy was taken. Significantly more LIF was produced by epithelial cells from late proliferative and early secretory endometrium compared with amounts produced by cells from early proliferative (P < 0.001) and late secretory (P < 0.01) endometrium. High doses of progesterone and oestradiol caused a small decrease in epithelial cell LIF production: the combined effect of progesterone and oestradiol (P < 0.01) was greater than the effect of either steroid alone (P < 0.05). The results show, for the first time, the capability of human endometrium to produce LIF in vivo. The fact that maximum LIF concentrations are present at implantation and that decreased concentrations occur in women with unexplained infertility suggest the importance of this cytokine in embryo implantation.      

MUC1 in secretory phase endometrium: expression in precisely dated biopsies and flushings from normal and recurrent miscarriage patients

MUC1 is a cell-surface and secretory product of endometrialepithelium. Immunohistoehemical studies carried out using twodifferent antibodies to the mucin-type tandem repeat regionof MUC1 indicate a cell-surface location in proliferative phaseglands, with intracellular deposits accumulating in the earlysecretory phase. Commencing 3–4 days after the luteinizinghormone (LH) peak and continuing into the late secretory phase,secretory MUC1 appears in gland lumens. Uterine flushings werecollected as a function of time after the LH peak and were analysedusing a two-site enzyme-linked immunosorbent assay for MUC1.Low but measurable concentrations were observed up to day 7,while on days 7–13 much higher values were obtained. Inwomen suffering from recurrent spontaneous miscarriage, theconcentration of MUC1 in flushings wassignificantly lower thanin the controls on day LH+10. Lower values were also observedon days 7 and 13. Reduced epithelial secretory function anda resultant change uterine fluid composition are features ofendometrium from recurrent miscarriage patients.     

Is the measurement of placental protein-14 and CA-125 in plasma and uterine flushings useful in the evaluation of peri-menopausal and post-menopausal bleeding?

In this prospective study, we examined the possible diagnosticvalue of the measurement of two endometrial proteins, placentalprotein-14 (PP14) and CA-125, in the evaluation of pre- andpost-menopausal bleeding. Concentrations of these two proteinswere measured in plasma and uterine flushings obtained from139 pre- and post-menopausal women with bleeding problems, and26 normal post-menopausal control women without bleeding. Endometrialbiopsy samples were also obtained for histological study. Concentrationsof PP14 in both the plasma and uterine flushings in post-menopausalwomen were significantly lower (P < 0.001) than those ofcontrol pre-menopausal women. In post-menopausal women, theconcentrations of PP14 (mean ± SEM) in both plasma andflushing were significantly higher (P < 0.001) in women withendometrial adenocarcinoma (46.9 ± 7.5 ng/ml plasma;3350 ± 1711 ng/ml flushing) than in the controls (7.6± 1.3 ng/ml plasma; 125 ± 27 ng/ml flushing) orin women with post-menopausal bleeding and atrophic endometrium(20.4 ± 2.1 ng/ml plasma; 453 ± 167 ng/ml flushing).In contrast CA-125 concentrations in plasma and flushings weresimilar in post-menopausal and pre-menopausal women. Plasmaconcentrations of CA-125 were higher in post-menopausal womenwith adenocarcinoma (29.1 ± 7.4 IU/ml) than in thosewith post-menopausal bleeding and atrophic endometrium (21.8± 2 IU/ml) (P < 0.05) or control post-menopausal subjects(16.1 ± 2.1 IU/ml) (P < 0.01). CA-125 concentrationsin uterine flushings were not significantly different in anygroup of post-menopausal women. The results show that concentrationsof PP14 are correlated more strongly to endometrial histopathologythan those of CA-125 in pre- and post-menopausal women. ElevatedPP14 concentrations are also associated with the presence ofendometrial adenocarcinoma and may have a potential to be usedas a marker for this disease.     

Concentrations of endometrial protein PP 14 and CA-125 in uterine flushings performed in natural and stimulated cycles

BACKGROUND: Impaired implantation in assisted reproduction cycles with high serum estradiol (E(2)) concentrations may be attributed to abnormal endometrial development. This study compared concentrations of endometrial proteins in uterine flushings of infertile patients between natural and stimulated cycles. METHODS: Patients received a standard regimen of ovarian stimulation. Seven days after the LH surge in natural cycles or the hCG injection in stimulated cycles, uterine flushings were performed by slowly injecting and aspirating normal saline through a paediatric Foley catheter. Natural cycles were considered as group A whereas stimulated cycles with serum E(2) <20 000 pmol/l and serum E(2) >20 000 pmol/l were classified as groups B and C respectively. PP 14 and CA-125 in uterine flushings were measured and expressed per total protein content. RESULTS: Concentrations of the total protein, PP 14 and CA-125 in the uterine flushings were similar among the three groups. PP 14 per total protein in the uterine flushings was significantly correlated with serum E(2) on the day of hCG (r = 0.459; P = 0.009) in natural cycles only but not in stimulated cycles. CONCLUSION: There was no significant difference between natural and stimulated cycles in concentrations of PP 14 and CA-125 in uterine flushings performed in the mid-luteal phase.     

Uterus and endometrium: Changes in nuclear morphology in the human endometrial glandular epithelium in women with unexplained infertility

In the light and electron microscopical study reported here,we document the structure of the nucleus, nucleolus and nuclearchannel system (NCS) in the uterine glandular epithelium inboth fertile and infertile women during the early luteal phase.Nuclear volume was found to be larger in the infertile groupat day 5 after the luteinizing hormone surge (LH + 5) comparedto day-matched fertile subjects. A two-way analysis of varianceperformed on nucleolar volume data from fertile and infertilewomen biopsied on days LH + 4, +5 and +6 revealed a significanteffect of condition but no effect of day or interaction. Nucleolarvolume decreased from day LH + 2 to day LH + 6 in the fertilegroup, the sharpest decrease occurring between day LH + 3 andday LH + 4. The largest mean volume of the NCS was found atday LH + 5 in the fertile group and day LH + 6 in the infertilegroup. The results suggest a delay in the development of thisorganelle in the infertile women. The present study has documentedalterations in the nuclei of uterine glandular cells from infertilepatients. In these infertile women, there is also a delayedelaboration of the secretory apparatus and this delay correlateswell with the delayed/reduced expression of a luteal-specificglyco-protein.     

MUC1 as a Cell Surface and Secretory Component of Endometrial Epithelium: Reduced Levels in Recurrent Miscarriage

The mucin MUC1 is a large, highly glycosylated, hormonally regulated product of endometrial glandular and luminal epithelium with both cell surface-associated and secreted isoforms. The abundance of mRNA coding for MUC1 increases about sixfold from the proliferative to the early secretory phase (Hey et al., J. Clin. Endocrinol. Metab. 78:337–342, 1994). Immunohistochemical studies show intracellular deposits accumulating in the early secretory phase followed by the release of MUC1 into gland lumens. The apical surface of luminal epithelium is strongly immunopositive in the early secretory phase. We have used a two site ELISA to measure MUC1 in uterine flushings as a function of time after the luteinising hormone (LH) peak. Low levels of secretory MUC1 are observed before day LH+7, while values on days LH+7-LH+13 are much higher. Using semi-quantitative immunohistochemical methods we have shown that in women suffering recurrent spontaneous miscarriage, mid secretory phase levels of MUC1 core protein and mucin-associated glycans are reduced (Serie et al., Fertil. Steril. 62:989–996, 1994). Similarly, lower core protein levels are observed in uterine flushings after day LH+7 in these women. Reduced epithelial secretory function and a resultant change in uterine fluid composition are features of endometrium from recurrent miscarriage patients.     

A prospective randomized controlled study comparing two doses of gestodene in cyclic combined HRT preparations on endometrial physiology

Postmenopausal women taking oestradiol 17-beta 2 mg daily were randomized to receive either 25 or 50 microg gestodene from day 17 to 28 of the cycle in a double-blind study. Placental protein P14 (PP14) and CA 125 concentrations in uterine flushing, endometrial morphology and irregular bleeding after 12 cycles of study were observed. Eleven and 12 women in the 25 and 50 microg groups respectively completed the study. There were no significant differences in pre-treatment biochemical and morphological indices between the groups. The median PP14 concentration increased from 332 to 5800 ng/ml (P < 0.001) and from 145 to 27 160 ng/ml (P < 0.001) in the 25 and 50 microg gestodene groups respectively. No between-group significant rise of PP14 was observed. Similarly, no significant change was seen between the initial and post-treatment concentrations of CA 125 for either group. All biopsies were atrophic at inception of the study, and both regimens produced secretory endometrial transformation in the majority of biopsies. No between-group difference was observed in the morphometric indices measured, or any significant correlation between the concentrations of PP14 or CA 125 and morphology. The mean number of days of withdrawal bleeding (3.8 and 4.2 days for 25 and 50 microg respectively) were similar. In conclusion, both regimens produced a significant rise in uterine flushing concentrations of PP14, but not CA 125. PP14 is a sensitive biochemical marker in the assessment of endometrial response to hormone replacement therapy.     

Placental protein 14 in cycles with normal and retarded endometrial differentiation

The purpose of this study was to investigate whether endometriumwith retarded development differs, functionally, from endometriumwith normal ’in-phase‘ development. Precisely timedendometrial biopsies were obtained from 24 women suffering fromunexplained infertility at 4, 7, 10 and 13 days following theluteinizing hormone(LH)surge. Frozen sections were labelledwith an anti-placental protein(pp)14 monoclonal antibody usingan avidin-biotin peroxidase technique and semi-quantificationof endometrial PP14 was performed using one-and two wat analysisof variance. Normal and retarded endometrium were identifiedin 16(group 1) and eight (group II)women respectively. Bothgroups demonstrated a significant increase of the area of precipitatemeasured for PP14 form day LH +13. However, two-way analysisof variance showed that endometrial PP14 was significantly(P< 0.05)lower in the retarded endometrium group at LH+10 andLH+13. Serum PP14 was also significantly lower(P< 0.01) inwomen withretarded endometrial development had significantlyhigher (P< 0.05)concentration of cumulative saliva progesteronefrom LH +3 to LH +5. This study indicates that there are functionaldifferences between normal and retarded endometrium. These differencesmay adversely affect uterine receptivity during implantationand the early placentation stage     

Prognostic value of the measurement of uterine natural killer cells in the endometrium of women with recurrent miscarriage

BACKGROUND: Studies in mice suggest that CD56 + uterine natural killer (uNK) cells play an important role in implantation. Studies in humans have described an increase in the number of uNK cells in the non-pregnant mid-secretory endometrium of women with unexplained recurrent miscarriage (RM). However, the predictive value of uNK cell number in the maintenance of pregnancy is controversial. METHODS: A blind retrospective study was undertaken. The percentage of stromal cells positive for CD56 was identified by immunocytochemistry in endometrial biopsies from 10 normal control women and 87 women with unexplained RM, of whom 51 became pregnant following biopsy. Biopsies were obtained on days LH + 7 to LH + 9. RESULTS: As in previous studies, the number of uNK cells in the 87 women with RM (mean 11.2% range 1.1-41.4%) was significantly higher (P = 0.013) than in the control women (mean 6.2% range 2.2-13.9%). No significance difference in uNK numbers was observed between 19 women who miscarried (mean 9.6% range 1.7-25.0%) and 32 women who had a live birth (mean 13.3% range 1.1-41.4%) in a subsequent pregnancy. CONCLUSIONS: In this study numbers of uNK cells in the peri-implantation endometrium of women with unexplained recurrent miscarriage did not predict subsequent pregnancy outcome.     

Markers of endometrial function in women with unexplained recurrent pregnancy loss: a comparison between morphologically normal and retarded endometrium

BACKGROUND: Endometrial defect, usually described as luteal phase defect (LPD), is associated with recurrent miscarriage. Recurrent miscarriage has also been associated with the abnormal expression of various molecules by endometrial cells. The aim of this study was to determine if any of these molecules or cells could be used to distinguish LPD from in-phase endometrium. METHODS: Immunocytochemistry was used to compare endometrial expression of CD45+, CD56+, CD3+ and CD4+ cells, leukaemia inhibitory factor, interleukin-6 and estrogen and progesterone receptors in precisely timed endometrial biopsies obtained between days LH+6 and LH+11 from recurrent miscarriage women with in-phase and retarded endometrium. RESULTS: In all samples there was a positive correlation between the number of CD45+ cells and LH day and a negative correlation between progesterone receptor and LIF expression and LH day. A significantly lower number (P<0.05) of CD56+ cells in peri-implantation endometrium and a decreased mid-cycle estrogen level (P<0.05) was seen in women with LPD compared to in-phase endometrium when single analysis was carried out. However, these differences were not significant after application of the Bonferroni correction for multiple analysis. CONCLUSIONS: The results are in line with previous associations observed between estrogen levels and LPD and suggest that the number of CD56+ cells is different in LPD and in-phase endometrium, although this could be due to delayed endometrial development in women with LPD. Interpretation must be cautious because these differences could have arisen by chance.     

Pregnancy: Serum concentrations of luteinizing hormone and progesterone in ovum recipients: their relationship with pregnancy and miscarriage

Our objective was to test the hypothesis that the associationbetween elevated luteinizing hormone (LH) concentrations andmiscarriage is mediated via an effect of LH on the maternalenvironment, rather than on the oocyte. The impact of maternalage, ovarian function, previous IVF attempts, therapeutic (buserelin)and hormonal (LH, oestradiol, progesterone) effects occurringon the day of zygote intra-Fallopian transfer (ZIFT) or embryotransfer, and of oocyte or embryo numbers, whether they werefresh or frozen, and their mode of transfer on the occurrenceof pregnancy and miscarriage following ovum donation (n = 57)were investigated. The cycles were divided by outcome into non-pregnant(n = 26), miscarriage (n = 19) and normal term pregnancy (n= 12). The circulating concentrations of LH were greater inmiscarriage cycles (P = 0.046) and cycles ending in pregnancy(P = 0.04) than in non-pregnant cycles, while the concentrationsof progesterone were greater in non-pregnant (P = 0.029) andmiscarriage (P = 0.015) cycles than in cycles ending in pregnancy.Frozen embryos were used more frequently in non-pregnant comparedto cycles ending in pregnancy (P = 0.016). Multiple regressionanalysis was used to investigate which factors are associatedwith miscarriage and identified progesterone concentrationsat the time of transfer as being the only significant variable(r = 0.48, F = 8.5, P = 0.007). The same method of analysiswas used to investigate which factors are associated with thefailure to conceive and identified previous IVF attempts (F= 5.8, P = 0.021), the presence of ovarian function (F = 5.7,P = 0.022), the use of frozen zygotes (F = 5.1, P = 0.029) andprogesterone concentrations (F = 5.9, P = 0.02), with an overallresult of r = 0.59, F = 5.2 and P = 0.002. In conclusion, highprogesterone concentrations were associated with the failureto conceive and miscarriage. In contrast, LH concentrationswere lower in women who failed to conceive but similar in pregnantwomen who did and did not miscarry. This suggests that the associationbetween elevated LH concentrations and infertility is via adirect effect of LH on the oocyte and an indirect effect, mediatedby elevated progesterone concentrations, on the endometrium.     

The effects of post-ovulatory administration of onapristone on the development of a secretory endometrium

The purpose of this study was to assess the ability of the anti-progestinonapristone administered in the immediate post-ovulatory periodto disrupt endometrial differentiation as a potential methodof fertility control. In all, 10 healthy female volunteers weregiven 400 mg onapristone 2 days after the mid-cycle luteinizinghormone (LH) surge in urine (LH+2), An endometrial biopsy wastaken 4 or 6 days after the LH surge (i.e. LH+4 or LH+6) ina control cycle and on the corresponding day of the treatmentcycle. Biopsies were assessed for histological dating and immuno-localizationof oestrogen receptors, progesterone receptors and 15-hydroxyprostaglandindehydrogenase (PGDH). On day LH+12, blood was taken for themeasurement of insulin-like growth factor binding protein-1(IGFBP-1) and placental protein 14 (PP14). Hormonal measurementsin blood and urine were used to monitor the effects on the menstrualcycle. In addition, the concentration of cortisol in plasmawas measured to determine if this dose of onapristone exertedsignificant anti-glucocorticoid activity. Treatment with onapristoneretarded the development of secretory changes within the endometriumwithout affecting the length of the luteal phase. Intense nuclearimmunostaining of oestrogen and progesterone receptors was evidentin glands and stroma after treatment, suggesting that the progesterone-dependentdown-regulation of steroid receptors was inhibited by the anti-progestin.Onapristone also affected the production of luteal phase endometrialproteins, as judged by the pronounced reduction in immunostainingof PGDH within the glands and the significant reduction in plasmaconcentrations of PP14. However, plasma concentrations of IGFBP-1did not differ between cycles. Onapristone did not appear toexert significant anti-glucocorticoid activity because concentrationsof cortisol were unaffected. These findings suggest that onapristonecould potentially be used as a method of post-ovulatory fertilitycontrol.     

Effect of low weekly doses of mifepristone on ovarian function and endometrial development

The effect of a low dose of mifepristone (RU486) on ovarianand endometrial function was studied in 14 healthy women. Thestudy included one control and two treatment cycles. Duringthe treatment cycles, either 2.5 mg (n = 9) or 5 mg (n = 5)of mifepristone was administered once weekly. The concentrationof ovarian steroids and luteinizing hormone (LH) in urine wasmeasured daily, cortisol in blood once weekly and glycodelin(placental protein 14; PP14) at the time of menstruation. Ovarianfunction was monitored by vaginal ultrasound. An endometrialbiopsy was taken in each cycle in the mid-luteal phase, basedon self-measurement of the LH peak, or on cycle day 22 if noLH peak could be detected. In the evaluation of the results,the outcome of the enzyme immunoassay of LH was used to datethe biopsy. Endometrial progesterone and oestrogen receptorsand Dolichus biflorus agglutmin (DBA) lectin binding were measured.Ovulation was not inhibited by treatment with mifepristone,and an LH peak could be determined in all control and treatmentcycles. However, in four subjects (one with the higher and threewith the lower dose) the follicular phase was prolonged by 6–13days. The duration of the luteal phase and the concentrationsof pregnanediol and oestrone glucuronide were not affected bytreatment A dose of 5 mg, and to a lesser extent 2.5 mg, mifepristoneonce weekly caused desynchron-ization of endometrial developmentEndometrial progesterone receptor, but not oestrogen receptor,concentration was significantly increased by the higher dose.A significant reduction in DBA-lectin binding and in serum glycodelinconcentrations was also found. Thus, low doses of mifepristonedo not inhibit ovulation but delay endometrial development andimpair secretory activity. Whether these effects are sufficientto prevent implantation remains to be established.     

Concentrations of placental protein 14 in uterine flushings from infertile women: validation of the collection technique and method of expression of results

Concentrations of various proteins in uterine flushings have been described as a direct method for assessment of the secretory activity of the endometrium. We investigated levels of the endometrial protein known as placental protein 14 (PP14) in flushings obtained from 271 infertile women. Under transvaginal ultrasonographic control, 2 ml of 0.154 M sodium chloride solution were injected into the uterine cavity and re-aspirated, five times. In contrast to previous studies the recovered volume of each flushing was not consistent (range: 0.05-2.1 ml); the volume varied significantly between serial samples obtained from an individual (P = 0.02, one-way ANOVA), different cycle days (P < 0.0001, one-way ANOVA) and women with bilaterally blocked versus patent Fallopian tubes (P < 0.05, Student's t-test). Concentrations of PP14 showed a better correlation with protein content (r = 0.506, P < 0.0001) than with the recovered volume (r = 0.087, P = 0.095). We therefore corrected PP14 concentrations for total protein content as an indicator of the efficiency of the flushing process. Corrected PP14 concentrations varied significantly relative to time since the onset of menstruation (P = 0.001, Kruskal Wallis ANOVA) with higher levels on days 1-8, as previously observed in plasma samples. No significant difference in PP14 levels was found with different causes of infertility. This study shows that uterine flushing is not a consistent process in women with differing physical characteristics and at varying times throughout the menstrual cycle.      

Peripheral CA 125 levels in patients with uterine fibroids.

CA 125, a marker of ovarian cancer, is also increased in otherwise normal women suffering from, for example, pelvic inflammatory disease, endometriosis and adenomyosis. The tissues suspected of producing CA 125 in normal women include the endometrium, the ovary and the peritoneum. This study was based on the hypothesis that uterine myomata would distend the peritoneum covering the uterus and thereby increase the peripheral levels of CA 125. To verify this hypothesis we measured CA 125 by an immunoradiometric assay in eight normal women every second day throughout the cycle and in 26 women with uterine fibroids before and after hysterectomy and at 8 and 12 weeks during gonadotrophin releasing hormone (GnRH) analogue therapy. In normal women no difference was observed between CA 125 levels in the follicular phase or in the luteal phase of the cycle. Over one-third (10/26) of the patients with uterine fibroids had increased (greater than 90th centile of the controls) levels of CA 125 before GnRH therapy or hysterectomy. Removal of the uterus or administration of GnRH significantly decreased peripheral concentrations of CA 125 to levels below those observed in normal women. Furthermore, a significant positive correlation was observed between the levels of CA 125 and the volume of myomata as assessed by ultrasound. We conclude that in those cases of uterine fibroids where CA 125 is increased, monitoring this parameter during GnRH therapy is a good indirect measurement of regression of myomata.     

Association of 14-bp insertion/deletion polymorphism of HLA-G gene with idiopathic recurrent miscarriages in infertility center patients in Yazd,Iran

HLA-G is supposed to play a pivotal role in tolerance of the semi-allogeneic graft in pregnancy by inhibiting the cytotoxic functions of T and NK cells. A 14-bp insertion and/or deletion polymorphism in exon-8 has a possible role in HLA-G expression. The present study analyzed the 14-bp insertion/deletion polymorphism in normal pregnancy and recurrent miscarriage patients in order to discover a possible correlation between the 14-bp polymorphism and recurrent miscarriage (RM). In this study, genomic DNA from 200 RM patients and 200 normal fertile control individuals using the routine salting out method were isolated. Exon-8 of HLA-G gene of the two groups were amplified using polymerase chain reaction and analyzed by electrophoresis on 10% non-denaturing polyacrylamide gel electrophoresis containing ethidium bromide and visualized under ultraviolet light. HLA-G allele frequencies and genotypes in RM women and the fertile control group were compared using a Chi-square test. The results showed that there was a difference in allelic frequencies of 14-bp insertion polymorphism between fertile controls and RM patients; the frequency of +14 bp/?14?bp heterozygotes was significantly higher in RM patients as compared with fertile controls. Furthermore, the frequency of +14-bp insertion allele was significantly higher in those with RM as compared with normal fertile controls. From the findings here, it was concluded that a 14-bp insertion/deletion polymorphism in exon 8 could play a possible role in recurrent miscarriages. These results might ultimately be of significance for clinicians and those involved in understanding infertility and RM.     

Endocrine abnormalities during the follicular phase in women with recurrent spontaneous abortion.

The frequency of endocrine abnormalities during the follicular phase in non-pregnant women with a history of recurrent abortion was investigated in a case-control study. A total of 42 consecutive women with recurrent spontaneous abortion (three or more consecutive abortions, mean +/- SD: 3.9 +/- 1.1 range 3-8) with no parental chromosome rearrangement or uterine abnormality were studied during the early follicular phase under standardized conditions. Serum concentrations of follicle stimulating hormone (FSH), luteinizing hormone (LH), prolactin, androstenedione, testosterone, dehydroepiandro-stenedione, 17-OH-progesterone, oestradiol, progesterone and thyroid stimulating hormone (TSH) were measured by commercially available radioimmunoassays. Controls were 42 nulligravid females with tubal or male factor infertility without miscarriage. Mean (SD) concentrations of prolactin and androstenedione were 14.2 +/- 6.7 ng/ml versus 10.5 +/- 3.5 ng/ml (95% CI 0.8-6.1) and 2.3 +/- 0.9 ng/ml versus 1.7 +/- 0.6 ng/ml (95% CI 0.2-0.9) in the study and control groups respectively. The other endocrine parameters were comparable in both groups. Obesity [BMI weight (kg)/height (m2) > or = 25] was more prevalent (23 versus 5 women, P = 0.0001) in the study than the control group. Recurrent spontaneous abortion is associated with abnormalities in prolactin and androgen secretion during the follicular phase, suggesting an endocrine aetiology in this disorder. Reduction of body weight and correction of hyperprolactinaemia and of hyperandrogenism may reduce the rate of miscarriage in a subsequent pregnancy in these women.     

Changes in basement membrane thickness in the human endometrium during the luteal phase of the menstrual cycle

We have examined aspects of the fine structure of the basal laminae associated with the luminal and glandular epithelium and small blood vessels in the human endometrium. Four short studies are presented and reviewed. Study 1 examined biopsies from 20 fertile women taken on days after the luteinizing hormone surge (LH): LH +2, 4, 6, 8 and 10. The basal lamina (both lamina densa and lucida) increased in thickness over the period studied. Study 2 again studied the glandular epithelium and examined the effect of RU486 (a progesterone receptor blocker) administered on day LH +3 and biopsied on day LH +6. The basal laminae were found to be the same as LH +2 control group but thinner than LH +6 control. Study 3 documented increased thickness of the basal laminae between LH +6, 8 and 13 in the luminal epithelium. The within-group coefficient of variation was 16% and 27% for LH +6 and LH +13 groups but only 2 % for LH +8. Study 4 demonstrated an increase in basal lamina thickness associated with small blood vessels between LH +6 and LH +10 in normal fertile women. The basal lamina provides the interface between epithelial and mesenchymal environments; changes in its structure can alter the phenotypic expression of the epithelia. It is one of the maternal barriers that must be transgressed by the trophoblast during implantation. Together, these combined studies provide quantitative baseline structural information on the electron microscopical appearance of the basal lamina during the luteal phase of the menstrual cycle.