Enzyme immunoassay of oestrogen and progesterone receptors in uterine and intrauterine tissue during human pregnancy and labour.

Oestrogen and progesterone receptors were studied in the non-pregnant state, in early pregnancy and at term using monoclonal antibody enzyme immunoassays. Receptors for both steroids were found in tissues from non-pregnant patients and patients in early pregnancy. At term oestrogen receptors were undetectable in all tissues studied. Progesterone receptors were undetectable in chorion, amnion and placenta at term, while present in extremely low concentrations in decidua and myometrium.

Metabolic profiles and lipoprotein lipid concentrations in non-obese and obese patients with polycystic ovarian disease.

Clinical parameters, androgen status and lipoprotein lipid profiles were assessed in 10 non-obese and 10 obese patients with polycystic ovarian disease (PCOD) and reference subjects matched for age, height and weight. Both obese and non-obese women with PCOD had significantly higher androgen levels when compared to the reference groups. When comparison of lipoprotein lipid profiles were made between groups, non-obese women with PCOD had significantly higher total cholesterol, triglycerides and LDL-cholesterol levels than non-obese reference subjects. Obese PCOD women manifested significantly higher total cholesterol, LDL-cholesterol, cholesterol/HDL, and LDL/HDL values than did obese reference subjects. Correlations between serum androgens and lipoprotein lipid concentrations in PCOD and normal women were unhelpful. Both non-obese and obese patients with PCOD had significantly higher systolic and diastolic blood pressures (BPs) than the reference groups. Thus, both non-obese and obese women with PCOD manifest hyperandrogenaemia which may result in a male pattern of lipoprotein lipid concentrations.

Glucose, insulin and C-peptide secretion in obese and non obese women with polycystic ovarian disease.

Plasma glucose, immunoreactive insulin (IRI) and C-peptide responses during oral glucose tolerance testing (OGTT) were evaluated in 10 non obese women with polycystic ovarian disease (NOB-PCOD) and 10 obese women with polycystic ovarian disease (OB-PCOD). Mean plasma glucose response at 120 minutes in OB-PCOD showed impaired glucose tolerance. Also in this group, 1 patient had frank diabetes mellitus, whilst 3 other patients had impaired glucose tolerance 1 NOB-PCOD patient had impaired glucose tolerance. Mean plasma glucose levels and mean incremental glucose areas were higher in the OB-PCOD at all time intervals and reached statistical significance at 60 and 90 minutes. Mean plasma IRI levels were also higher in OB-PCOD at all time intervals, and reached statistically significant higher levels at 0, 60 and 90 minutes. Mean serum C-peptide valves were also higher at all time intervals in OB-PCOD. The relationship between acanthosis nigricans, obesity and PCOD was also analysed. It is evident from this study that obesity has a significant negative impact on the overall carbohydrate status in women with PCOD.

125I-insulin binding to circulating erythrocytes, monocytes and cultured fibroblasts in non-obese patients with polycystic ovarian disease.

The insulin receptor status of circulating erythrocytes, monocytes and cultured fibroblasts were studied in non-obese Indian women with polycystic ovarian disease (PCOD) with no clinical evidence of acanthosis nigricans and age, height and weight matched reference subjects. The women with polycystic ovaries had decreased mean maximum specific binding to erythrocytes (PCOD 7.4 +/- 0.6%, normal women 11.5 +/- 0.3%; p less than 0.0001) and monocytes (PCOD 2.4 +/- 0.3%, normal women 4.1 +/- 0.4%; p less than 0.003) when compared to the normal women. This decreased binding was attributed to a change in both receptor number and affinity. 125I-insulin binding to cultured fibroblasts revealed similar mean maximum specific binding and affinity values in both groups studied. Although further work is necessary to exclude genetic or post-receptor defects, it is likely that an environmental factor is implicated in the decreased 125I-insulin binding to erythrocytes and monocytes.

Changes in the binding characteristics of oestrogen and progesterone in uterine and intrauterine tissue with the progression of human pregnancy.

The binding characteristics of oestrogen and progesterone in intrauterine tissue were studied in the non-pregnant state and throughout pregnancy using sucrose density gradient centrifugation and titration analyses. High affinity receptors for these steroids which were present in high concentrations in tissue from non-pregnant patients and patients in early pregnancy, decreased with the advance of pregnancy to undetectable levels at term. Non-saturable binding to progesterone detected in term tissue was identified as albumin and cortisol binding globulin.

Insulin and C-peptide secretion in non-obese patients with polycystic ovarian disease.

Plasma glucose, immunoreactive insulin (IRI) and C-peptide responses during an oral glucose tolerance test (oGTT) were assessed in 11 non-obese patients with polycystic ovarian disease (PCOD) and 11 reference subjects matched for age, height and weight. Also, 6 patients with PCOD and 6 normal women were subjected to intravenous glucose tolerance testing (ivGTT) On oGTT, all subjects exhibited normal glucose tolerance; however, PCOD patients had significantly higher mean plasma glucose levels at 30, 60, 90 and 120 min and higher mean incremental glucose areas. In addition the patients with polycystic ovaries showed higher mean basal IRI and C-peptide levels, higher mean glucose stimulated IRI and C-peptide levels and higher mean incremental IRI and C-peptide values. The molar ratios of C-peptide/IRI were significantly lower in the PCOD group at all time intervals after glucose stimulation when compared to the normal women. During ivGTT, there were significantly higher mean glucose levels at 5, 40, 50 and 60 min in the PCOD group when compared to the reference group. The IRI response to intravenous glucose in the PCOD women was similar to the reference group. The findings on oGTT suggest that non-obese patients with PCOD have increased pancreatic IRI secretion as well as impaired hepatic extraction of the hormone.

Comparison of human chorionic gonadotrophin concentrations in the sera of patients with normal and abnormal pregnancy measured by radio-immunoassay and immunoradiometric assay.

Human chorionic gonadotrophin (HCG) concentrations were measured in the serum of 40 patients with normal pregnancy, 18 with ectopic pregnancy, 36 with incomplete abortions and 24 patients with gestational trophoblastic disease using a radio-immunoassay (RIA) and immunoradiometric assay (IRMA). There were no significant differences in HCG concentrations measured by the two assays with regard to the first three groups but the patients with gestational trophoblastic disease had consistently higher results on RIA, probably reflecting increased circulating free beta-subunit of HCG. It was concluded that measurement of HCG using IRMA for the intact molecule gives equivalent results to RIA in patients with normal and ectopic pregnancy as well as after abortion. However, monitoring of patients with choriocarcinoma and hydatidiform mole requires an assay that detects free beta-HCG.

Spectrum of thyroid function abnormalities in gestational trophoblastic neoplasia.

The relative influences of the putative thyroid stimulator, pregnancy state and non-thyroidal illnesses, were assessed in 52 patients with gestational trophoblastic neoplasia (GTN). In this study biochemical thyroid status was assessed with the aid of thyrotrophin (TSH) measurements by a sensitive immunoradiometric assay and measurement of total and free thyroid hormone concentrations. The reference group consisted of 63 patients with normal pregnancy. Thyroid function in the GTN patients was also compared with that in 36 female patients with Graves' disease and 34 female patients with non-thyroidal illnesses. We found the major influence on thyroid functions in Group I ('Hyperthyroid') patients with GTN to be the presence of the thyroid stimulator, while non-thyroidal illnesses played an important role in Group II ('High T4 syndrome') and III ('euthyroid') patients with GTN. Thyroid function was also stimulated in Group II patients. The influence of pregnancy was seen in all three groups of patients with GTN, who had higher TBG concentrations than non-pregnant patients. We conclude that there is a spectrum of thyroid function abnormalities in GTN, and that thyroid function in an individual patient is determined by the relative influence of the thyroid stimulator, non-thyroidal illnesses and the pregnancy.

Serum progesterone in the diagnosis of ectopic pregnancy: a valuable diagnostic test?

The value of a single serum progesterone (P) assay in the diagnostic work-up of suspected ectopic pregnancy was investigated in 89 patients with ectopic pregnancy and 27 patients with incomplete abortion. Reference values for P in the blood were obtained from 77 patients with normal intrauterine pregnancies in the first trimester. With the use of a discriminatory level of 63 nmol/l (20 ng/ml), sensitivity of 92%, specificity of 84%, positive predictive index of 90%, and negative predictive index of 87% were achieved. Addition of pelvic ultrasound further improved the diagnostic accuracy. It is concluded that serum P measurement offers a valuable adjunct to existing methods of diagnosis of ectopic pregnancy.

Serial oxytocin levels in amniotic fluid and maternal plasma during normal and induced labour.

Oxytocin was measured by a specific and sensitive radioimmunoassay in plasma and amniotic fluid after extraction with Sep-Pak cartridges in patients undergoing elective caesarean sections, normal labour and labour induced with oxytocin infusion or prostaglandins. In maternal plasma, levels of oxytocin correlated with the period of gestation; concentrations at term were significantly higher than in the first two trimesters. Maternal plasma levels of oxytocin before the onset of contractions were not significantly different from those at the onset of spontaneous labour or at full cervical dilatation. Levels of oxytocin in patients induced with oxytocin were not statistically different from levels observed in spontaneous labour. Amniotic fluid oxytocin levels were not significantly different between the groups either at the onset of labour or immediately before delivery. Umbilical arterio-venous differences in oxytocin were present in all groups except the prostaglandin-induced group. Increased prostaglandins associated with the onset of normal labour are probably not a consequence of changes in oxytocin concentrations.

The SimulTRAC FT4/TSH assay evaluated as a first-line thyroid-function test.

We evaluated the SimulTRAC FT4 57Co/TSH 125I dual-isotope assay for the simultaneous measurement of free thyroxin (FT4) by radioimmunoassay analog techniques and of thyrotropin (TSH) by immunoradiometry. Inter- and intra-assay CVs were less than 10% over the entire range tested except for 15.9% at the lowest FT4 concentration. Results obtained by the SimulTRAC assay allowed complete differentiation of 85 hyperthyroid patients and 35 hypothyroid patients from normal subjects. However, such estimations of FT4 or TSH concentrations occasionally were misleading for assessing thyroid status in various clinical conditions. We conclude that the SimulTRAC assay has the same inherent disadvantages possessed by FT4 analog and TSH immunoradiometric assays; however, where results of one of the simultaneous assays may be misleading, the results provided by the other may indicate the underlying pathology without requiring an additional assay.

Impaired corpus luteum function in ectopic pregnancy cannot be explained by altered human chorionic gonadotropin.

We studied the cause of the low serum progesterone, 17 beta-estradiol, and 17-hydroxyprogesterone levels that occur in women with an ectopic pregnancy. Only women who had been amenorrheic for less than 8 weeks were studied in order to assess corpus luteum rather than placental biosynthesis of these steroids; each woman with an ectopic pregnancy was matched to a woman with a normal intrauterine pregnancy on the basis of serum intact hCG levels within 10% of one another to obviate the influence of different levels of this luteotropic hormone. Every woman with an ectopic pregnancy had lower serum progesterone, estradiol, and 17-hydroxyprogesterone levels than her matched normal pregnant pairmate (median values: progesterone, 27.9 vs. 83.5 mmol/L; estradiol, 0.36 vs. 1.79 nmol/L; 17-hydroxyprogesterone, 4.95 vs. 22.1 nmol/L, respectively; all P less than 0.002). The ratios of intact hCG, measured by immunoradiometric assay, to hCG, measured by a hCG beta-specific RIA, were similar in the two groups. Serum hCG bioactivity was assayed by measuring the ability of serum to stimulate testosterone secretion from mouse Leydig cells. The mean biological to intact immunological hCG ratios were 2.06 +/- 1.39 (+/- SD) for ectopic pregnancy and 1.91 +/- 0.81 for normal pregnancy (P greater than 0.05). The biological hCG to immunoreactive hCG beta ratios were 1.98 +/- 0.75 and 2.02 +/- 0.82, respectively. Serum hCG from both groups of women stimulated cAMP generation by testicular cells similarly. We conclude that the lower serum steroid levels in women with ectopic pregnancy cannot be explained by altered hCG bioactivity. The lower steroid levels may thus reflect a primary defect of the corpus luteum, absence of another stimulator of ovarian steroid biosynthesis, or more subtle alterations in hCG glycosylation which are important in vivo but not assessed by the in vitro bioassay.

Autoantibodies to thyroxin and triiodothyronine.

Two clinically euthyroid patients with multinodular goiter were found to have high "free" thyroxin (Amerlex-M, Amersham and Coat-a-Count, DPC) and triiodothyronine (Amerlex-M) concentrations (FT4 and FT3, respectively). The presence of antibodies to T4 and T3 was confirmed by the finding that polyethylene glycol precipitated a far greater proportion of radioactivity when radiolabeled FT4 or FT3 analog (Amerlex-M) was incubated with serum from these patients than was true for normal subjects. With this method we could not demonstrate antibodies to thyroid hormones in 116 healthy volunteers. Of 101 hyperthyroid patients tested, one had antibodies to T4 but none had antibodies to T3. One patient had antibodies to T4, and one to T3, of 36 hypothyroid patients tested. All patients with thyroid hormone antibodies also demonstrated antithyroglobulin antibodies (measured immunoradiometrically). Evidently, the presence of thyroid hormone antibodies should be suspected when results of thyroid-function tests are discordant with the clinical state, and we suggest that measurement of thyrotropin by an assay with improved detection limits will aid in correctly determining thyroid status.

Blood or urine measurement of human chorionic gonadotropin for detection of ectopic pregnancy? A comparative study of quantitative and qualitative methods in both fluids.

One hundred seventy-five patients with features of ectopic pregnancy were studied, of whom 95 were subsequently shown to have an ectopic pregnancy. Paired blood and urine samples were assayed for human chorionic gonadotropin (hCG) by two radioimmunoassays, one immunoradiometric assay for intact hCG and an immunoradiometric assay for free beta subunit that also detects the "beta core" of hCG in urine. Qualitative testing was also performed using the Tandem Icon method of immunoconcentration on a membrane. The quantitative results for intact hCG showed an approximately unitary relationship between concentrations in both fluids, with a close correlation (r = 0.84, gradient = 1.01). The qualitative tests for hCG in both serum and urine were positive in all patients with ectopic pregnancy. The Tandem Icon is equally useful in blood and urine, whereas quantitative assays are more reliable in the blood. Quantitation of urinary hCG is not recommended because of the variable dilution of the glycoprotein in this fluid.

Evidence for increased absorption of glucose and decreased hepatic extraction of insulin in South African Indians.

It has been previously demonstrated that Indians have an exaggerated insulin response to oral glucose when compared to Africans. In an attempt to determine the reasons for this phenomenon matched groups of Indian and African volunteers were subjected to oral and intravenous glucose tolerance testing. It was concluded that the increased absorption of glucose and reduced hepatic extraction of insulin contributed to the hyperinsulinism in Indians.

Detection of urinary human chorionic gonadotropin by rapid immunoconcentration method is the first-line test for suspected ectopic pregnancy.

Nine hundred nine patients with suspected ectopic pregnancy were tested for human chorionic gonadotropin (hCG) using two qualitative assays: the Tandem Icon urine assay (Hybritech, San Diego, CA) and a serum radioimmunoassay (RIA) employing a positive cut-off of 25 IU/l. Pregnancy status was determined by clinical or histologic examination, or detection of hCG or its metabolites in four quantitative immunoassays: two RIAs and two immunoradiometric assays (IRMAs). Both the Tandem Icon and the qualitative RIA detected all 71 patients with ectopic pregnancy. The predictive indices of the Icon were 100% for a positive result and 99.6% for a negative result, and those of the RIA were 96.7 and 99.6%, respectively. The Tandem Icon thus fulfils the need for a simple, rapid, and sensitive method for hCG in the detection of patients with suspected ectopic pregnancy.

Chloroquine resistance in Plasmodium falciparum in Natal.

Out of 110 cultures of Plasmodium falciparum obtained from infected patients in Natal 18 were found to be resistant to chloroquine by in vitro tests: 2 cultures showed schizont development in wells containing 24 pmol chloroquine; in 16 schizont development was present in culture wells containing 48 pmol chloroquine. Seventy-two patients with P. falciparum malaria who apparently did not respond to adequate oral chloroquine therapy were investigated. All responded to one or more treatments with pyrimethamine/sulphadoxine (Fansidar; Roche). It would seem that all the patients were infected in areas not controlled by the South African health authorities.

Amniotic membrane production of prostaglandin F2 alpha is reduced in dysfunctional human labor: results of in vivo and in vitro studies.

Mobilization of arachidonic acid from glycerophospholipids and prostaglandin (PG) release from fetal membranes were studied in women with dysfunctional labor in the absence of cephalopelvic disproportion or fetal malposition. Using superfusion of intact amnion and chorion, we found a slight decrease in PGE and a more significant decrease in PGF release by the amniotic side of the fetal membrane obtained from women with dysfunctional labor compared to that in women with normal labor (PGE: normal labor, 2992 pg/cm2.h; dysfunctional labor, 1846 pg/cm2.h; P less than 0.05; PGF: normal labor, 662 pg/cm2.h; dysfunctional labor, 204 pg/cm2.h; P less than 0.02). Release of both prostanoids was significantly greater from the amniotic side in tissues obtained after labor compared to that in prelabor tissue. Analysis of arachidonic acid (by gas liquid chromatography) and phospholipid content (by two-dimensional thin layer chromatography) confirmed metabolic disposal of arachidonic acid from the amnion after the onset of labor. However, no difference in either phospholipid or phospholipase A2-releasable arachidonic acid of individual phospholipid classes was found in amnion tissue from women with normal and dysfunctional labor, suggesting similar activities of phospholipase A2 in these two groups. The finding of decreased free and phospholipase A2-releasable arachidonic acid of the total lipid extract of the amnion of women with dysfunctional labor could suggest further metabolic exhaustion of the substrate or failure of liberation of this fatty acid from glycerophospholipids by enzymes other than phospholipase A2, such as phospholipase C or diacyl and monoacylglycerolipases.