Phospholipase activity in the endometrium of women with normal menstrual blood loss and women with proven ovulatory menorrhagia

The activity of phospholipase A2 types 1 and 2 and phospholipase C was measured in the endometrium of women with ovulatory menorrhagia and in those with normal menstrual blood loss. In both groups of subjects phospholipase A2 type 1 activity was significantly higher in the secretory phase than in the proliferative phase (P less than 0.001). The median activity (pmol/mg protein/min) for the proliferative phase was 27.6 in normal subjects and 40.4 in women with ovulatory menorrhagia and for the secretory phase the median activity was 144.5 in normal women and 138.1 in women with ovulatory menorrhagia. There was no difference between the two groups of women at either stage of the cycle. Phospholipase A2 type 2 activity was also higher in the secretory phase than in the proliferative phase (P less than 0.05 for normal subjects and P less than 0.001 for women with menorrhagia). The median activity (pmol/mg protein/min) for the proliferative phase was 94.4 (normal subjects) and 56.6 (women with menorrhagia) and for the secretory phase 148.3 (normal subjects) and 142.5 (women with menorrhagia). The activity of phospholipase A2 type 2 was significantly lower in the proliferative phase of women with ovulatory menorrhagia compared with normal subjects (P less than 0.05). Phospholipase C activity (nmol/mg protein/min) was significantly higher in women with ovulatory menorrhagia (median 8.2) compared with women with normal blood loss (median 5.5) (P less than 0.01).     

Primary and myoma-associated menorrhagia: role of prostaglandins and effects of ibuprofen

Summary. The release of 6-keto-prostaglandin F_1α(6-keto-PGF_1α), a metabolite of prostacyclin (PGI₂) and thromboxane B2 (TxB2), a metabolite of thromboxane A2 (TxA2), was estimated in endometrial biopsies taken from 12 menorrhagic and 12 healthy women during the luteal phase of the cycle. The releases of 6-keto-PGF_1α and TxB2 were normal, but the ratio TxB2/6-keto-PGF_1α was inversely related to menstrual blood loss in women with measured menstrual blood loss exceeding 70 ml. In the second part of the study, 24 women with excessive menstrual bleeding (13 with primary menorrhagia, 10 with uterine fibro-myomas, one with haemostatic factor VIII deficiency) were treated at random with ibuprofen (600mg/day and 1200mg/day) and with a placebo. Ibuprofen 1200 mg/day reduced (P<0.01) median blood loss from 146 ml (range 71–374 ml) to 110 ml (30–288 ml) in primary menorrhagia but had no effect on blood loss in women with uterine fibroids and factor VIII deficiency. Blood loss was normal in six women and was not affected by ibuprofen. Thus, our data suggest that there is a PGI₂ dominance in the endometrium of patients with menorrhagia. In addition, primary, but neither fibromyoma nor coagulation defect-associated menorrhagia, can be treated by ibuprofen.     

Relationships between circulating leptin concentrations and other hormonal parameters in obese and non-obese women with polycystic ovary syndrome

Aim: To clarify the role of leptin in women with polycystic ovary syndrome (PCOS), we analyzed whether serum leptin levels correlate with other hormonal parameters in obese and non-obese women with PCOS.
Methods: We studied 20 obese (body mass index, BM ≥25 kg/m²) and 20 non-obese (BMI <25 kg/m²) women with PCOS diagnosed by the existence of menstrual disturbance, elevated serum level of luteinizing hormone (LH) with normal follicle-stimulating hormone (FSH) and the characteristic polycystic appearance of the ovaries on transvaginal ultrasound images. Blood samples for LH, FSH, estradiol, testosterone (T), androstenedione (Δ₄) and leptin were obtained, and the relationships between variables were examined by calculating Spearman correlation coefficients.
Results: Mean levels of leptin, T and Δ₄ in obese PCOS women were significantly higher than those in non-obese PCOS women, but this was not the case for BMI, bodyweight and waist to hip ratio. In all the 40 PCOS women considered together, there were significant positive correlations of leptin with BMI, waist to hip ratio, and Δ₄ levels. However, in each group separately, serum leptin levels in obese PCOS women correlated only with BMI and bodyweight, whereas serum leptin levels in non-obese PCOS women correlated with serum A4 levels.
Conclusion: Although further study is needed to assess the role of leptin on ovarian function in non-obese women with PCOS, present findings do not support the fact that leptin is involved in the development of hormonal abnormalities in obese women with PCOS. (Reprod Med Biol 2002; 1 : 49–54)     

The activity of calcium dependent and calcium independent phospholipase A2 in normal endometrium and in endometrium from women suffering from menorrhagia and polycystic ovary syndrome

The activity of 2 phospholipase A2 enzymes, PLA2(i) and PLA2(ii) was measured in endometrium in women with regular menstrual cycles without evidence of pathology and in those complaining of menstrual disturbances. There was a significant 4-fold increase in PLA2(i) activity in secretory phase endometrium (mean +/- SD: 32.7 +/- 9.5 pmol per mg protein/minute) compared to that of the proliferative and menstrual phases (9.5 +/- 4.9 and 6.1 +/- 2.6 pmol per mg protein/minute, respectively) but PLA2(ii) activity was variable and not related to the stage of the cycle (range: 4.0-97.0, 18.7-110.3 and 0.1-85.5 pmol per mg protein/minute for proliferative, secretory and menstrual phases, respectively). There was no significant difference between normal subjects and those with menorrhagia with respect to the mean activities of either isoenzyme at any stage of the cycle. Women with polycystic ovary syndrome (PCO) had markedly higher endometrial PLA2(ii) activity than normal subjects. The evidence of this study suggests that PLA2(ii) is not implicated in unexplained menorrhagia, but our preliminary findings indicate that the high level of PLA2(ii) activity found in the endometrium of women with PCO might be a marker of abnormal endometrial function.     

Uteroplacental blood flow and placental vascular endothelial growth factor in normotensive and pre-eclamptic pregnancy

Objective To determine whether placental vascular endothelial growth factor (VEGF) is increased in pre-eclampsia.
Design Prospective cohort study.
Setting Royal Prince Alfred Hospital, Sydney, Australia.
Sample Eleven normotensive women and eight women with pre-eclampsia matched for age and gestation.
Methods Uterine artery Doppler ultrasound flow velocity profiles were recorded in the third trimester and resistance index calculated as (V_s-V_d)/V_s (V_s= peak systolic flow velocity, V_d= end diastolic flow velocity). Placental tissue at delivery was examined for VEGF distribution with avidin-biotin-peroxidase immunohistochemistry.
Results Uterine resistance index [median (range)] was significantly increased in pre-eclamptic women (normotensive: 0.42 (0.36–0.51); pre-eclampsia: 0.59 (0.40–0.75);   P = 0.005  ). Notching of the uterine artery waveform, consistent with a high resistance circulation, was evident in early diastole in five women with pre-eclampsia but only one normotensive woman (   P = 0.013  ). Placental VEGF was increased in women with pre-eclampsia in the decidual trophoblast (normotensive: 34% (4–59) cells stained for VEGF; pre-eclampsia: 58% (15–95);   P = 0.033  ) and in the villous syncytiotrophoblast (normotensive: VEGF count 1.4 arbitrary units (1.1–2.1); pre-eclampsia: 1.8 arbitrary units (1.4–2.2);   P = 0.041  ). Analysis indicated that uterine artery resistance index was directly correlated with placental VEGF staining, mean arterial pressure and birthweight.
Conclusions Abnormal uterine artery Doppler ultrasound flow velocity profiles in pre-eclampsia indicate increased uteroplacental resistance. The associated increase in placental VEGF may represent a compensatory mechanism attempting to restore blood flow towards normal.     

Blood levels of proteinase inhibitors in pregnancy

Summary. Plasma volume, serum α₂-macroglobulin, α₁-antitrypsin, Cï inactivator and α₂-antiplasmin, and plasma antithrombin III were measured in 10 pregnant women at gestation periods of 12–14 and 37–38 weeks. The proteinase inhibitors were also measured in 10 non-pregnant healthy women. There was a significant increase in the α₁-antitrypsin concentration and significant decreases in α₂-macroglobulin and Cï inactivator, but the total circulating quantity of all the proteinase inhibitors was significantly increased.     

Integrin expression in cervical carcinoma

Heatley MK, Corke K. Integrin expression in cervical carcinoma. Int J Gynecol Cancer 1998; 8: 203–206.
Integrin expression was studied in a series of 36 cervical carcinomas using antibodies to integrins αvβ₅, α₃, β₁ and β₄. Integrins αvβ₅, α₃ and β₁ were localized to the cell membrane in well differentiated (0/13, 2/4 and 4/14 cases) and moderately differentiated adenocarcinomas (0/6, 1/5 and 1/6 cases, respectively) and in adenosquamous (2/6, 2/5 and 3/6) well differentiated (1/4, 0/3, and 3/5) and moderately differentiated (1/2, 1/3 and 1/3) squamous cell carcinomas. Two antibodies (AA3 and 439-B) located integrin β₄ to the cell membranes and cytoplasm of well (13/13 and 10/12 cases) and moderately differentiated (6/7 and 5/6 cases) adenocarcinomas, cases of adenosquamous carcinoma (6/6 and 5/6 cases), and well (4/4 and 3/4 cases) and moderately (3/3 and 3/4 cases) differentiated squamous cell carcinomas. In conclusion, integrin expression was identified most frequently with the antibodies to β₄ which was localized to cervical carcinomas of varying grades and histological types. Immuno-staining was identified less frequently with the antibodies to αvβ₅, α₃ and β₁.     

Serum markers for Down's syndrome in women who have had in vitro fertilisation: implications for antenatal screening

To examine the Down's syndrome screening positive rate among in vitro fertilisation (IVF) pregnancies, we measured second trimester serum marker levels in singleton IVF pregnancies (cases) and in five non–IVF pregnancies (controls) matched to each case for gestational age, age of mother, and duration of storage of the serum sample. There were 151 IVF pregnancies in which alpha fetoprotein, unconjugated oestriol (uE₃), free β–human chorionic gonadotrophin (hCG) and total hCG were measured, 104 IVF pregnancies in which free α-hCG was measured, and 39 IVF pregnancies in which inhibin A was measured. Median uE₃ levels were 6% lower (   P = 0.003  ), median free β–hCG 9% higher (   P = 0.024  ), and median total hCG 14% higher (   P = 0.026  ) in IVF pregnancies compared with controls. The screen positive rate in the IVF pregnancies (28%) was about twice as high as that in controls (17%). High hCG levels may be explained by progesterone remaining high in IVF pregnancies. The low uE₃ levels remain unexplained. In Down's syndrome screening in IVF pregnancies hCG and uE₃ values should be adjusted to avoid the high screen positive rate.     

Immunohistological localization of pregnancyassociated endometrial α2-globulin (α2-PEG) in endometrial adenocarcinoma and effect of medroxyprogesterone acetate

Summary. Endometrium from postmenopausal women with endometrial adenocarcinoma was examined immunohistochemically using a monoclonal antibody to pregnancy-associated endometrial α₂-globulin (α₂-PEG), the major secretory protein of the glandular epithelium during the late luteal phase of the menstrual cycle and early pregnancy. Specimens were obtained at initial diagnostic curettage and at hysterectomy after medroxyprogesterone acetate (MPA) therapy. α₂-PEG was not detected in any malignant tissue irrespective of histological differentiation. Non-malignant endometrium obtained in association with malignant tissue was negative for α₂-PEG before treatment although after MPA therapy all specimens obtained exhibited marked α₂-PEG localization in glands. In four specimens endogenous alkaline phosphatase was observed consistently only in the malignant endometrium. Malignant endometrium does not appear to synthesize α₂-PEG nor is its synthesis induced by an oral progestogen, so that it does not represent a useful marker for endometrial carcinoma. Non-malignant endometrium in postmenopausal women appears to be fully capable of α₂-PEG production after stimulation with an oral progestogen.     

Human placental lactogen and pregnancy-specific β1-glycoprotein in pregnant diabetic women

Summary. Serum placental lactogen (hPL) and pregnancy-specific β₁ glycoprotein (SP₁) concentrations were measured in 16 insulin-dependent pregnant diabetic women and in 16 non-diabetic control subjects matched for placental weight. hPL concentrations were found to be significantly higher in diabetics with placental weight >90th centile compared with those in (i) non-diabetic controls with placental weight >90th centile and (ii) diabetics with placental weights <90th centile. SP_l concentrations were higher in both diabetic groups compared with those in their respective nondiabetic controls, but no difference existed between the two diabetic groups. In the control subjects infant birthweights were lower in the women with small placentae, but no such difference was observed between the two diabetic groups.     

Uterine artery embolisation for symptomatic fibroids: the effect of the large uterus on outcome

Objective   The aim of this study was to evaluate the efficacy of uterine artery embolisation (UAE) in myomatous uteri larger than 24 week's gestation (780 cm³).
Design   Prospective case contro study.
Setting   Universitas Hospital, University of the Free State, Bloemfontein, South Africa.
Population   Sixty-one women, who underwent UAE, were included in the study. The study group comprised of 12 women with uteri ≥780 cm³ and the control group 49 women with uteri <780 cm³.
Methods   UAE was performed and the difference in outcome for the two groups was determined at 12 months.
Main outcome measure   Symptomatic improvement with embolisation of the large uterus.
Results   Reduction of dysmenorrhoea, menorrhagia and pressure effects was similar for both groups. The median reduction in uterine volume (pre- to post-embolisation) was 188 cm³ (range 28–2038 cm³) with a 95% CI for the median difference for paired data of 146.5 and 236. Only 66% of the study group had, however, a reduction in volume to <780 cm³. The complication rates were similar for the two groups with regards to post-embolisation syndrome, fibroid slough, haematoma formation, infection, hysterectomy and failure to embolise. Satisfaction was similar between the two groups, with 91% of women satisfied with the procedure.
Conclusion   The large uterus does not decrease UAE's efficacy. Although 33.3% of the study group still had a uterus of ≥780 cm³, symptom reduction was still similar for both groups. Women may thus still be left with a large uterine volume but without symptoms. This must be taken into consideration when counselling women with an extremely large uterus for UAE.     

Oocyte donation: a review

Summary. Oocyte donation provides an option for achieving pregnancy in women lacking functioning gonads, or in whom IVF techniques have failed to harvest adequate oocytes, or those who do not wish to use their own gametes because of hereditary disease. In agonadal women, artificial menstrual cycles are required before proceeding to gamete donation. A fixed cyclical steroid replacement schedule of oestradiol (E₂) valerate and progesterone (P₄) pessaries was initially used, but the need for synchrony between donor and recipient cycles, and the narrow window for implantation limited the transfer of fresh embryos. Donorrecipient cycle asynchrony can be overcome by using frozen-thawed embryos, or by extending the follicular phase in the recipient to widen the transfer window. Twenty-two pregnancies have now been achieved by the Monash/Epworth group, resulting in the birth of 13 healthy Infants. There were no statistically significant differences in pregnancy rates (per transfer) between transfers in natural cycles (14%, four pregnancies) and steroid replacement cycles (24%, 16 pregnancies). Five pregnancies (36%) were established in women treated with 2 mg of E₂ daily 13–18 days before embryo transfer with P, starting on the day of or the day tollowing oocyte retrieval. E₂ was continued for a median of 85 days (range 49–110) and P₄ for a median of 86 days (range 49–133) after the supposed last menstrual period. All but one delivery was by caesarean section. There were no perinatal deaths and no ectopic pregnancies.     

Reduced fetal exposure to aspirin using a novel controlled-release preparation in normotensive and hypertensive pregnancies

Objectives To examine the fetal effects of a novel controlled-release, low dose aspirin preparation in normal and hypertensive pregnancies.
Design Random double-blind study. Participants assigned to receive conventional formulation aspirin (75 mg), controlled-release low dose aspirin (75 mg), or matching placebo.
Setting National Maternity Hospital, Dublin.
Participants Eighteen women with an uncomplicated pregnancy and 18 women with preeclampsia.
Main outcome measures Urine was analysed for metabolites of thromboxane and prostacyclin by gas chromatography, mass spectrometry. Serum thromboxane B₂, was determined in maternal and cord blood.
Results Both aspirin preparations reduced maternal serum thromboxane B₂, by 95% and induced similar reductions in the urinary 11-dehydro-thromboxane B₂, a major metabolite of thromboxane A₂ in vivo. In contrast, neither preparation altered urinary 2,3–dinor-6-keto PGF_1α, the major metabolite of prostacyclin. Despite their similar effects in the mothers, the two aspirin preparations differed in their effects on the fetus. While both suppressed cord fetal thromboxane B₂, this was significantly (  P < 0.005  ) less for the controlled-release preparation (210 ± 42 ng/ml for placebo vs 109 ± 22 ng/ml for controlled-release aspirin and 44 ± 9 ng/ml for regular oral aspirin).
Conclusions At equivalent maternal suppression of serum thromboxane B₂, a controlled aspirin release preparation results in lower fetal exposure than regular oral aspirin.     

Expression of integrin adhesion molecules in normal ovary and epithelial ovarian tumors

The metastatic potential of a solid tumor is dependent upon its ability to interact with the extracellular matrix. The integrin superfamily is a group of proteins that are fundamental in such interactions and play a major role in cell-cell and cell-matrix adhesion. Localization of the integrin proteins was performed in normal ovary, primary epithelial ovarian tumors and metastatic tumor cells in ascitic samples. Expression of α₁, α₃, α₆ and β₄ was observed on normal ovarian epithelium with variable expression of α₅. Loss of α₁ expression by malignant cells in the primary tumors was noted. β₄, a component of the laminin receptor which was strongly expressed by both normal ovary and solid tumor, was absent from the ascitic tumor cells in the majority of cases. There was an associated loss of α₆ expression, indicating a deficiency of hemidesmosomes in the ascitic tumor cells. This alteration of integrin expression by metastatic malignant epithelial ovarian tumor cells may therefore represent one important mechanism by which metastatic disease occurs.     

Serum protein pattern in normal pregnancy with special reference to acute-phase reactants

Summary. The concentrations of acute-phase protein reactants, total protein, albumin and globulin fractions were measured throughout normal pregnancy in 27 women. α₁-Antitrypsin and caeruloplasmin concentrations increased gradually to reach their highest levels in the third trimester. Orosomucoid and haptoglobin showed similar patterns: higher levels in the first and third trimester with a decline around 24 weeks gestation. C-Reactive protein showed levels similar to those of non-pregnant healthy individuals (< 5 mg/1) throughout pregnancy. α₁,-, α₂ and β-Globulin concentrations increased from the first trimester towards term. γ-Globulin concentration changed little during gestation. The data obtained provide reference ranges for serum proteins in healthy pregnancy.     

Association of gestational diabetes with abnormal maternal vascular endothelial function

Objective To evaluate vascular endothelial function in isolated small arteries from women with gestational diabetes.
Methods Small subcutaneous arteries (mean luminal diameter ∼ 250μm) were dissected from biopsies obtained at caesarean section in 14 normotensive women with gestational diabetes and in 18 normotensive nondiabetic pregnant women. Vascular function was determined after mounting the arteries on a small vessel myograph.
Results Pre-constricted arteries from gestational diabetic pregnant women demonstrated poor relaxation to acetylcholine, an endothelium-dependent vasodilator (pEC₅₀, mean [SE], 6.98 [0.10] vs normal pregnant, 7.28 [0.08],   P < 0.03  ; % maximum relaxation, median [range], 88.2 [42.4–994] vs normal pregnant 94.2 [71.8–100.0],   P < 0.01  ). In the presence of indomethacin relaxation to acetylcholine was similar in both groups suggesting a deficiency in dilator prostaglandin synthesis in the arteries from the diabetic women. The nitric oxide synthase inhibitor N -monomethyl-L-arginine further reduced sensitivity of arteries to acetylcholine but to a similar degree in both normal pregnant and gestational diabetic women. Relaxation to sodium nitroprusside, an indicator of sensitivity of the vascular smooth muscle to nitric oxide, was similar in both groups.
Conclusions Maternal vascular endothelial dysfunction may contribute to the increased incidence of cardiovascular disorders in women with gestational diabetes.     

Use of a super active luteinizing hormone releasing hormone (LHRH) agonist in the treatment of menorrhagia

Summary. A luteinizing hormone releasing hormone agonist, d-Ser(But)⁶des Gly¹⁰-LHRH ethylamide, was administered intranasally to four women with unexplained ovulatory menorrhagia. Dramatic reductions in excessive menstrual blood losses were induced without significant side-effects, although the pattern of loss returned to pretreatment levels within 2 months of ceasing therapy.     

Phytoestrogens and reproductive biology

Phytoestrogens are naturally occurring plant substances that can either mimic or antagonize the action of endogenous estrogens. This is because of the similarity of the functional structure of phytoestrogens and endogenous estrogens. In premenopausal women, phytoestrogen intake might induce a decrease in luteinizing hormone, follicle-stimulating hormone and estradiol (E₂), which are associated with a longer follicular phase. The circulating transport protein, sex hormone-binding globulin, is increased, resulting in less cellular availability of E₂. Phytoestrogens inhibit the activities of E₂ synthetic enzymes through adenylate cyclase and tyrosine kinase cascades. This might decrease of risk of hormone dependent cancers. A phytoestrogen-rich diet might reproduce normal body composition, affecting the course of polycystic ovary syndrome (PCOS). Some herbs used in traditional Japanese medicine contain phytoestrogens that influence endogenous hormone levels to directly regulate the pituitary-ovarian system, in particular, the chemotactic effects on ovaries. (Reprod Med Biol 2005; 4 : 225 –229)     

Low dose 25 mg oestradiol implants and 1 mg norethisterone as continuous combined hormone therapy: a prospective study

The anxiety regarding no-bleed regimens is that breakthrough bleeding and endometrial hyperplasia may occur. We aimed to demonstrate that 25 mg oestradiol implants can be adequately opposed by a low dose of progestogen protecting against osteoporosis. Twenty-two patients were recruited to the study. The mean age was 62 years and body mass index of 26.5. Median oestradiol rose from 77 pmol/L at baseline to 275 pmol/L at one year. Median endometrial thickness remained unchanged at 4 mm and only two women withdrew with bleeding problems. There was one case of proliferative endometrium at one year—all others samples were either atrophic or secretory. Lumbar bone density (L2–L4) rose significantly from  0.939 to 0.992 g/cm²  (  +5.6%  ,   P = 0.005  ) and the total femoral density rose from  0.872 to 0.890 g/cm² (+2.1%)  . Bone formation markers increased significantly (serum type 1 procollagen C terminal peptide,  P1CP = 112–114, P = 0.0376  ) and bone resorption fell (serum type 1 collagen C terminal telopeptide,  1CTP = 3.0–2.9, P = 0.2863  ). E25 implants and low dose progestogen appear to avoid endometrial hyperplasia and bleeding problems while increasing bone density.