Simple, rapid and semiquantitative enzyme-linked immunoassay for detection of the preovulatory LH excretion in the urine.
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Biomedical subjects
Publications and source records attributed to O Tanizawa.
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A fluorometric liquid chromatographic method was developed for measurements of unconjugated oestradiol and oestriol in the serum of pregnant women. The serum samples are injected directly into the apparatus and pass to a pretreatment column, where oestrogens are adsorbed while hydrophilic components such as proteins and carbohydrates are not. The oestrogens then pass into a separation column containing a new type of polymer gel. The mobile phase consists of an acetonitrile-water mixture, and separation is achieved by a reversed-phase mechanism. The eluent is monitored for fluorescence. Data on the reproducibility and recovery by this method and the correlation of values with those obtained by radioimmunoassay are reported. Results on the increases of oestradiol and oestriol in the serum during pregnancy are also reported.
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Various assay methods for detection of thyroid hormones in human milk were evaluated in recovery and dilution experiments after which the concentrations of thyroxine (T4) and 3,5,3'-triiodothyronine (T3) were measured and compared with those in serum. The effect of breast feeding on pituitary thyroid function of normal babies also was studied. Competitive protein-binding analysis (CPBA) was found to be unsuitable for measurement of T4 in milk. T4 was not detected in samples of human milk by four radioimmunoassays (RIA), although more than 100% of T4 was recovered in the assays. RIA (double antibody-ANS system) seemed to be reliable for detection of T3 in milk, judging from recovery and dilution experiments. T3 was detectable in all samples obtained 1-4 months postpartum. The T3 concentration in milk was not correlated with protein concentration or daily volume. The concentration of T3 in milk was lower than that in serum and the mean ratio of serum T3 to milk T3 was 2.8 +/- 1.7 (mean +/- S.D.). No correlation was observed between the T3 concentration or daily T3 excretion in milk and the T3 concentration in serum. The total amount of T3 excreted in milk was estimated as only 5-1000 ng/day. The serum levels of thyrotropin, T4, free T4 and T3 were not significantly different between breast-fed and bottle-fed babies. These results indicate that T3 excretion in milk cannot be explained by simple diffusion from the blood into the mother's milk and that breast feeding has no influence on the pituitary thyroid axis of normal babies.
Maternal plasma oxytocin levels during pregnancy and labor were measured by a sensitive and specific radio-immunoassay. The mean plasma oxytocin concentration in maternal peripheral plasma showed a gradual rise towards term, reaching 12.5 microU/ml in term subjects without labor contractions. This level was not significantly different from the mean value in the 1st stage of normal spontaneous labor (12.6 microU/ml). Brief fluctuations in plasma oxytocin levels of considerable degree, and sometimes spurt release, were observed in labor and at term, but not in the mid-trimester, in serial samples collected at short intervals (about 10 sec). In normal spontaneous labor, no obvious relation was observed between the plasma oxytocin level and the uterine contraction period, and no significant difference was found between the mean value in uterine contraction periods (12.1 microU/ml) and that in relaxation periods (11.5 microU/ml).
In 4 women with anencephalic fetuses, the oxytocin level was measured in the maternal plasma in the second stage of labor and in the umbilical artery (UA) and umbilical vein (UV) by radioimmunoassay (RIA). In these cases, the mean maternal OT level was 13.4 +/- 8.0 microU/ml (mean +/- S.E.), which was not significantly different from the OT levels in cases of normal spontaneous birth and elective cesarean section. The UA and UV plasma OT levels in these cases were below the detectable level (less than 2 microU/ml), whereas in normal spontaneous births, they were 18.2 +/- 4.9 microU/ml and 12.2 +/- 2.3 microU/ml, respectively. The OT level in the UA was higher than that in the UV. There was no difference between the mean OT levels in patients with, versus those without contractions, but the plasma OT levels in the second stage of labor were higher than those in the first stage. In normal pregnancies the plasma OT concentration increased during parturition. These results suggest that placental transfer of maternal plasma OT to the fetal circulation is limited, and that in cases of anencephalic fetuses, maternal labor does not necessarily require fetal OT as a trigger or for its maintenance.
Serum free thyroxin (FT4) in normal nonpregnant and pregnant subjects was measured by radioimmunoassay (RIA) with Amerlex FT4 RIA (Amersham International) and LiquiSol FT4 RIA (Damon Diagnostics) kits. Amerlex FT4 values in serum from pregnant women were lower than those in serum from nonpregnant women, but LiquiSol FT4 values were similar in serum from both groups. Amerlex FT4 values were directly correlated with the concentrations of albumin in serum and inversely correlated with those of thyroxin-binding globulin, but not with prealbumin concentrations. No significant correlations were observed between LiquiSol FT4 values and serum concentrations of thyroxin-binding proteins. Amerlex FT4 values were normal in patients with excess, deficient, or decreased thyroxin-binding globulin. Albumin added to serum samples increased Amerlex FT4 values but not LiquiSol FT4 values. Albumin inhibited the binding of labeled thyroxin analog to the solid-phase thyroxin antibody. These data indicate that the albumin concentration influences FT4 values as measured by an RIA involving a thyroxin analog and that Amerlex FT4 values should be carefully interpreted when the patient has an abnormal concentration of serum albumin.
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A newly synthesized 16, 16-dimethyl trans delta 2 PGE1 methyl ester (ONO 802) was clinically applied in the form of a vaginal suppository for therapeutic abortion of second trimester pregnancies. Its effects were studied in a double-blind test comparing it with an inactive placebo suppository. The study was conducted at 12 Japanese university hospitals. The number of patients was 125 in total, i.e. 63 receiving ONO 802 vaginal suppository (containing 1.0 mg ONO 802) and 62 receiving inactive placebo suppository. ONO 802 was more effective than placebo with a success rate of 87% (complete (71%) and incomplete (16%) abortions). The onset of uterine contractions was observed in 154.3+/-18.1 min. and the onset of uterine bleeding in 323.6+/-41.0 min. The expulsion of the fetus and placenta was observed in 955.4+/-97.0 and 961.6+/-97.0 min., respectively. The cervix dilating effect of ONO 802 was observed in 63.5% of the patients at 3 hours after the start of administration. Nausea, vomiting, abdominal cramps, diarrhea and pyrexia were noticed. However, all these side effects were transient and mild, requiring no treatment. No abnormality was observed in the puerperal course, duration of uterine bleeding or onset of subsequent menstruation following the therapeutic abortion. Therefore, the present study demonstrated that ONO 802 vaginal suppository was an effective and valuable drug for therapeutic abortion of second trimester pregnancies.
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The effect of pregnancy on the clinical course of Graves' disease was examined by studies on 41 pregnancies in 35 patients with Graves' disease, who were considered to be in a state of remission or near remission and were not receiving antithyroid drugs, during and after delivery. Eighteen of the 41 cases (44%) showed transient increases in the serum free T4 index (FT4 index) during weeks 10--15 of pregnancy, but normal thyroid function in the second and third trimesters. Similar transient increases in the serum free T3 index (FT3 index) were observed in early pregnancy in these patients. These early increases in the FT4 and FT3 indexes were specific to Graves' disease and were not observed in Hashimoto's disease. Two to 4 months postpartum, 32 cases (78%) developed various degrees of thyrotoxicosis, which was divided into 3 types: 1) persistent thyrotoxicosis with high radioactive iodine (RAIU) (10 cases), 2) transient thyrotoxicosis with normal or high RAIU) (10 cases), and 3) destruction-induced thyrotoxicosis with low RAIU (12 cases). An increase in the FT4 index in early pregnancy was significantly (P less than 0.001) associated with relapse of stimulation-induced thyrotoxicosis of either the persistent or transient type. Patients who developed destruction-induced thyrotoxicosis after delivery had significantly higher titers of antithyroid microsomal antibodies (P less than 0.001) and a longer euthyroid period before pregnancy (P less than 0.01) than patients who had recurrent persistent thyrotoxicosis. These data indicate that Graves' disease is aggravated in early pregnancy and after delivery and ameliorates in the latter half of pregnancy. Postpartum relapse of persistent hyperthyroidism could be predicted from an early increase in the FT4 index during pregnancy.
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In this sensitive, simple method for measuring "free thyroxin" (FT4) in eluates of dried blood spots on filter paper by use of a radioimmunoassay kit (Amerlex Free T4 RIA), the measurable range of FT4 is 1.8 to 57 ng/L (equivalent to the concentration in serum), or 7 to 237 fg/tube. The mean coefficients of variation for within assay-within spots, within assay-between spots, and between assays were 5.3%, 5.0%, and 6.2%, respectively. FT4 in blood spotted on filter paper is stable for at least a month when dried and kept at either -20 degrees C, 4 degrees C, room temperature (about 25 degrees C), or 37 degrees C. The results for FT4 in dried blood spots correlated closely with the free-T4 concentration in serum (r = 0.99). The method can be used to differentiate cases of primary and secondary hypothyroidism from normal subjects and those with subnormal thyroxin-binding globulin. This method may be useful in screening for congenital hypothyroidism, because sample-retesting is not necessary.
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