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M W Seif

Publications and source records attributed to M W Seif.

35 records · Page 2Linked to original sources
PubMed1994

Endometrial differentiation in the peri-implantation phase of women with recurrent miscarriage: a morphological and immunohistochemical study.

OBJECTIVES: To study endometrial differentiation in the peri-implantation phase of women with recurrent miscarriage and to compare the results with endometrium of normal fertile women. DESIGN: A prospective study of endometrial specimens precisely timed from the LH surge, using traditional histologic dating (Noyes' criteria), quantitative histologic measurement (morphometric analysis), and immunohistochemical techniques. RESULTS: Fifteen of 25 (60%) subjects in the recurrent miscarriage group had retarded endometrial development in the peri-implantation period as monitored by morphometry. The recurrent miscarriage group showed reduced levels of four mucin-related secretory epitopes, and greater reductions were associated with morphological retardation. Normal differentiation was observed in all of the 14 subjects in the control group. CONCLUSIONS: Women with idiopathic recurrent pregnancy loss may be divided into two distinct subgroups on the basis of their endometrial response in the peri-implantation period. Precisely timed endometrial biopsy should be incorporated in the investigation of recurrent miscarriage.

Abortion, Habitual↗
PubMed1994

Expression of two isoforms of CD44 in human endometrium.

The distribution of the cell-surface adhesion glycoprotein CD44 in human endometrium was examined by immunofluorescence using six monoclonal antibodies to epitopes common to all forms of the molecule, and by reverse transcription-polymerase chain reaction (RT-PCR). Immunoreactivity was observed throughout the menstrual cycle in stroma, vessels, glandular, and luminal epithelium. Variations in staining intensity were observed, especially in the epithelial compartment. CD44 was also expressed strongly by decidualized stromal cells of first-trimester pregnancy. No systematic variation of immunoreactivity was observed with stages of the normal cycle, but a fraction (25%) of the specimens lacked reactivity in the epithelium. To determine the molecular size of the epithelial isoform, an immunoprecipitation technique was developed using surface-radioiodinated, detergent-extracted glands. This indicated the presence at the cell surface of a single dominant CD44E species with an approximate molecular mass of 130 kDa. RT-PCR was used to investigate the isoforms present in whole endometrial tissue, isolated gland fragments, and Ishikawa endometrial carcinoma cells. Complementary DNA produced from total endometrial mRNA was PCR-amplified across the splice junction between exons 5 and 15. Transcripts corresponding to the hyaluronate receptor CD44H as well as a larger isoform were identified. CD44H was absent, or very scarce, in cDNA from purified gland epithelium. In contrast, Ishikawa cells expressed this form abundantly. The glands and Ishikawa cells also expressed CD44E containing sequences encoded by exons 12, 13, and 14. These data demonstrate the presence of CD44 in human endometrium and decidua, and show that different isoforms of CD44 are associated with tissue compartments in which different functional roles can be anticipated.

Base Sequence↗
PubMed1994

Age related changes in the female reproductive cycle.

OBJECTIVE: To investigate the effect of age on the control of the reproductive cycle in regularly menstruating women. DESIGN: In this prospective study all women were seen regularly during both the follicular and luteal phases. Ultrasound scanning was used to confirm ovulation and measurements were taken to assess follicular development and endometrial thickness. Serum gonadotrophin levels on day 4 of the cycle, the maximum periovulatory oestradiol and progesterone on the seventh day after ovulation were measured and compared in different age groups. SUBJECTS & SETTING: Fifty-six healthy, regularly menstruating women were recruited from volunteers amongst staff of the University Hospital of South Manchester. RESULTS: Ovulation occurred later in the cycle for older women, with mean follicular phase length increasing from 13.9 days (in group 21-25 years) to 15.9 days (in group 37-45 years (P < 0.05). The mean of the maximum follicular diameter prior to rupture was significantly smaller in the older women: 16.7 mm in the 37 to 45 years old group compared with 19.6 mm, 21.6 mm and 21.3 mm in the 21 to 25, 26 to 31 and 32 to 36 years old age groups, respectively (P < 0.001). The maximum thickness of the endometrium in the luteal phase was greatest for older women: 15.9 mm in the age group 37 to 45 years compared with 12.1 mm in the age group 21 to 25 years (P < 0.001). Serum gonadotrophin concentrations during menses were higher with increased age; mean follicle stimulating hormone was 4.8 iu/l in the age group 21 to 25 years, and 8.5 iu/l in the age group 37 to 45 years (P < 0.001). Mean luteinising hormone was 4.5 iu/l in age group 21 to 25 years and 7.21 iu/l in age group 37 to 45 years (P < 0.001). Mean ovarian steroid concentrations were no different. CONCLUSIONS: These data illustrate significant age related differences in the pituitary-ovarian axis and endometrial thickness. This has implications for the management of older women in assisted reproduction and fertility control programmes.

Adult↗
PubMed1994

The polymorphic epithelial mucin MUC1 in human endometrium is regulated with maximal expression in the implantation phase.

After ovulation, progesterone stimulates a temporally regulated secretory transformation in human endometrial epithelium. Using a combination of immunohistochemistry, and Western and Northern blotting, we demonstrate that 1) the polymorphic epithelial mucin MUC1 is secreted by human endometrial epithelium; 2) low levels of both mRNA and core protein are present in the preovulatory phase of the menstrual cycle; 3) mRNA levels increase several-fold after ovulation, consistent with transcriptional regulation by progesterone; 4) there is an increase in translation product in postovulatory endometrium; and 5) the tandem repeat domain of the MUC-1 polypeptide is glycosylated in endometrium.

Antibodies, Monoclonal↗
PubMed1994

Ovulation induction and endometrial steroid receptors.

The endometrial morphology, endometrial steroid receptors and serum steroid hormone concentrations have been studied in 22 infertile women participating in an in-vitro fertilization, gamete intra-Fallopian transfer programme, including nine cases following treatment with gonadotrophin-releasing hormone analogue/human menopausal gonadotrophin/human chorionic gonadotrophin. All patients had normal ovulatory function before treatment and satisfactory response to ovulation induction. Endometrial biopsies were taken in spontaneous and treatment cycles on the fourth day after ovulation had been detected by ultrasound scanning, when endometrial receptors were measured using immunohistochemistry. Histological examination of biopsies in spontaneous cycles showed the majority (20/22) to be 'in-phase', while in two cases luteal phase defect was diagnosed. After ovulation induction, all the biopsies were still morphologically 'in-phase', although a significant reduction had occurred in the nuclear receptor level in both the glands and stroma for both progesterone receptors (gland P = 0.030, stroma P = 0.012 using microscopic analysis; gland P = 0.020, stroma P < 0.001 using a cell analysis system) and oestrogen receptors (gland P = 0.017, stroma P = 0.002 using direct microscopic analysis). This suggests that a reduction in steroid receptors in the endometrium occurs after ovulation induction in the presence of supraphysiological amounts of steroids, which is not associated with detectable morphological changes.

Adult↗
PubMed1992

Endometrium in in-vitro fertilization cycles: morphological and functional differentiation in the implantation phase.

Secretory differentiation of endometrium after multiple follicular stimulation using gonadotrophin releasing hormone and human menopausal gonadotrophin has been studied both histologically and immunohistochemically in 30 women undergoing in-vitro fertilization treatment. None had embryo transfer. Patients were randomly allocated to receive luteal phase support with a single dose of human chorionic gonadotrophin. The latter failed to produce any significant enhancement of endometrial structure or secretions. Appropriate glandular morphology was present in a greater proportion of those who were successfully stimulated than those who responded poorly. However, defective secretion of the cycle-dependent component studied, using monoclonal antibody D9B1, was demonstrated in two-thirds of cases regardless of the ovarian response. Early vascular maturation in the stroma was a common finding, and was thus considered as a feature of structural modulation of these endometria.

Antibodies, Monoclonal↗
PubMed1991

The effects of the antiprogesterone RU486 (Mifepristone) on an endometrial secretory glycan: an immunocytochemical study.

OBJECTIVE: To examine the effects of progesterone (P) receptor blockade by RU486 (Mifepristone; Roussel-Uclaf, Paris, France) on a secretory endometrial glycan recognized by monoclonal antibody D9B1. DESIGN: Retrospective comparison of endometrial biopsies from treated and untreated women from 2 to 8 days after the luteinizing peak (LH) peak. SETTING: Infertility clinic, Jessop Hospital for Women, Sheffield. PATIENTS: Twenty-two normal fertile women received the RU486. A control group of 44 normal fertile women were also assessed. INTERVENTIONS: RU486 was administered to 22 normal women during the first half of the luteal phase and an endometrial biopsy examined 3 days later. MAIN OUTCOME MEASURES: Immunohistochemistry was used to assess the production and secretion of the D9B1 epitope. RESULTS: When the drug was given 2 days after the LH peak, it prevented appearance of the epitope. When RU486 was administered 5 days after the LH peak, epitope already present in gland cells was subsequently secreted. CONCLUSIONS: These data suggest that production of the sialo-oligosaccharide is P-dependent, but secretion through established intracellular pathways is P-independent.

Adult↗
PubMed1990

An endometrial factor in unexplained infertility.

OBJECTIVE: To study a group of women with unexplained infertility to see whether they have a defect that is intrinsic to the endometrium. DESIGN: Evaluation of the functional response of the endometrium by examining endometrial biopsy specimens using immunohistochemical methods in a group of women with unexplained infertility and in a control group of women with normal fertility. PATIENTS: 27 Women with unexplained infertility (average age 33.2); median duration of infertility five years. A control group of 44 women with normal fertility (average age 33.8) who were requesting sterilisation or reversal of sterilisation. SETTING: Infertility clinic, Jessop Hospital for Women, Sheffield. INTERVENTION: Secretory phase endometrial biopsy specimens were taken, with informed consent, as an outpatient procedure. MAIN OUTCOME MEASURES: Immunohistochemistry with monoclonal antibody D9B1, was used to assess the production and secretion of an oligosaccharide epitope produced by endometrial gland cells between two and seven days after the luteinising hormone surge. A reflected light measuring system was used to assess the amount of epitope within the gland cells, and in the gland lumen. RESULTS: In the control group of women, mean reflected light measurements at the cell base and cell apex peaked at three and five days after the luteinising hormone surge respectively, and in the gland lumen the epitope accumulated rapidly from three days, reaching a peak at seven days. In the women with infertility the peaks of epitope at the cell base and cell apex were lower, broader, and delayed in onset, and the build up of epitope in the gland lumen was retarded. The synthesis and secretion of the epitope in the women with infertility was therefore significantly reduced and delayed, even in the presence of normal concentrations of circulating progesterone. CONCLUSIONS: The results suggest that a primary dysfunction of the endometrium might be associated with hitherto unexplained infertility.

Adult↗
PubMed1990

Menstrual-cycle-dependent expression of keratan sulphate in human endometrium.

Immunochemical methods have been used to detect and characterize two classes of polypeptide-associated keratan sulphate (KS) in epithelial secretions from human endometrium. Monoclonal antibody D9B1 binds to a hormonally regulated sialylated epitope associated with KS in a high relative molecular mass (250,000-350,000) component that bands as a doublet in SDS/PAGE. These KS chain(s) are sensitive to keratanase, endo-beta-galactosidase and N-glycanase. A second, more highly sulphated, type of KS is also present, that is resistant to all three enzymes. This can be detected using monoclonal antibody 5D4. It is present throughout the menstrual cycle and is associated principally with a component of Mr 140,000. Thus secretory KS contributes to the environment of the implanting embryo, may be used as a molecular index of endometrial function and could be important in the establishment of pregnancy.

Animals↗
PubMed1989

A novel approach for monitoring the endometrial cycle and detecting ovulation.

We report the presence of a cycle-dependent sialoglycoprotein in the endometrium. A monoclonal antibody (D9B1) to this glycoprotein has been derived and used to study tissue from 24 women with normal menstrual cycles. Results obtained with peroxidase immunohistochemistry suggest a highly significant variation in concentration of the glycoprotein, which is absent in the proliferative phase and present at maximal levels in the early secretory phase. The amount of antigen then diminishes slowly through the latter part of the secretory phase. The glycoprotein is produced in epithelial cells of glands and uterine lumen before being secreted across the apical cell surface. The secretory response is uniform in different areas of the tissue and within individual glands. However, considerable differences in secretory activity can be observed between adjacent glands in any part of the endometrium. Binding of the antibody is shown to be a new and novel parameter in characterization and standardization of the normal function of endometrium in response to ovarian hormones.

Antibodies, Monoclonal↗
PubMed1989

The effect of the intrauterine contraceptive device on endometrial secretory function: a possible mode of action.

The effect of the intrauterine contraceptive device (IUD) on the composition of endometrial secretion has been studied using the monoclonal antibody D9B1. The antibody binds to a polypeptide-associated oligosaccharide epitope that is secreted by endometrial epithelium in the secretory phase with a maximum around the time of implantation. In endometria affected by the IUD in situ, a significant reduction in epitope expression has been observed using immunohistochemistry. This defect can occur against a background of normal secretory differentiation and in the expectation of normal ovarian stimulus. The data provide new insight into the contraceptive mechanisms of action of the IUD.

Adult↗
PubMed1989

Luteal phase defect: the possibility of an immunohistochemical diagnosis.

Monoclonal antibody D9B1 has been shown to bind to a carbohydrate epitope associated with high molecular weight secretory sialoglycoprotein(s) of human endometrium. The authors demonstrate that, in a group of 28 patients diagnosed on the basis of histopathologic assessment as exhibiting luteal phase defect, 68% reveal significantly diminished expression of the epitope. Furthermore, histologic assessment of the secretory activity in single glands does not correlate simply with the level of expression of the epitope. Of specimens from patients complaining of infertility, 92% showed defective production of the D9B1 epitope. Thus, D9B1 immunohistochemistry provides a new molecular criterion for defective endometrial function.

Antibodies, Monoclonal↗
PubMed1989

The endometrial cycle: the expression of a secretory component correlated with the luteinizing hormone peak.

This paper reports the pattern of production and secretion of a secretory phase epitope in the endometrium of 44 normal fertile women. Peroxidase immunochemistry was used to detect the monoclonal antibody D9B1, which binds to a peptide-associated sialylated oligosaccharide, in endometrial biopsies all chronologically dated from the luteinizing hormone (LH) peak. During the proliferative phase, the epitope was shown to be absent from tissue sections. It first made its appearance within gland cells 2 days after the LH peak. By LH+3 a rapid accumulation of the D9B1 epitope was noted in the base of the cell, below the nucleus. On subsequent days, increasing amounts of the antigen were detected in the apical cytoplasm, apparently in a more concentrated form than in the basal cell zone. On day LH+6, around the time of implantation, secretory vesicles in the cell apex were discharged into the gland lumen where the glycoconjugate finally accumulated. This immunohistochemical approach introduces a new parameter for evaluation of endometrial function and could be used to improve the accuracy of histological assessment of the endometrial cycle.

Adolescent↗
PubMed1988

Expression of a secretory product by microvillous and ciliated cells of the human endometrial epithelium in vivo and in vitro.

A monoclonal antibody which identifies a component of post-ovulatory endometrial secretions is now shown to be expressed within the cytoplasm and on the cell surface of both microvillous and ciliated epithelial cells. A glandular explantation model was developed in order to study the 'carry over' of this secretion to the regenerative phase endometrium. A loss of cytoplasmic antigen was observed in vitro. However, it was retained on the cell surface in a fashion consistent with its expression at the time of explantation. Mosaicism of expression of this secretory component occurs throughout the secretory-phase and is particularly pronounced at the time of transition from proliferative to secretory phase. It is concluded that both ciliated and microvillous epithelial cells produce a post-ovulatory secretory component which may be retained on the cell surface in the absence of hormonal stimulation.

Cells, Cultured↗
PubMed1987

A monoclonal antibody to a cell surface determinant in human endometrial epithelium: stage-specific expression in the menstrual cycle.

A monoclonal antibody (CC25) was obtained after immunization of mice with intact glandular epithelial cells from secretory phase endometrium. Here we report a preliminary immunohistologic study in the endometrium of 29 patients in different phases of the menstrual cycle and early pregnancy. In immunofluorescence, CC25 binds to a basolaterally oriented epithelial cell surface antigen that is absent during the proliferative phase and appears suddenly in both glandular and uterine surface locations soon after ovulation. In mid and late secretory phase, the level of expression diminishes slowly. The epitope is absent from glandular epithelial cells in first-trimester decidua. However, it is associated with vascular smooth muscle cells in endometrium and decidua. CC25 promises to be a useful reagent for the analysis of endometrial function during the menstrual cycle.

Animals↗
PubMed1985

Basally located epithelial cell surface component identified by a novel monoclonal antibody technique.

Tubular aggregates of glandular epithelial cells (gland fragments) were isolated from human endometrium by collagenase digestion of surrounding stroma, thus exposing the basal surfaces of the cells. Using these aggregates as immunogen, monoclonal antibodies could be derived that recognized basally located antigens. One such antibody, G71, is described, that binds to a basal epithelial cell antigen present in a variety of human epithelia. Epitope-bearing molecules in the range Mr 60 000-180 000 are present in two of the tissues studied, amnion and endometrium. The epitope is associated with areas of epithelial cell-extracellular matrix contact.

Amnion↗