Basic evaluation of highly sensitive thyroid stimulating hormone immunoradiometric assay kits

Usefulness of three kinds of TSH kits by immunoradiometric assay (IRMA) was evaluated. They were able to measure low levels (less than 0.1 microIU/ml) in serum thyroid stimulating hormone (TSH) with incubation of short time (4 hours). In particular, RIABEAD II kit had a highly specific affinity for TSH and the normal range (+/- 2 S.D.) using it showed from 0.20 to 3.50 microIU/ml in 150 normal subjects. In patients with hyperthyroidism and in patients with hypothyroidism, the values of TSH were lower and higher than those of normal subjects, respectively. Another kits showed similar results. These results indicate that these TSH-IRMA kits are useful to evaluate serum TSH levels exactly.     

Radioimmunoassay of cholylglycine in serum (author's transl)

Serum levels of cholylglycines (CG) were determined by radioimmunoassay and that of total bile acids (TBA) by enzymatic method. In normal subjects, serum levels of CG, TBA and CG/TBA ratio were 0.6 +/- 0.4 micronM, 7 +/- 2 micronM and 0.08 +/- 0.06, respectively. They were increased markedly in acute hepatitis and moderately in chronic hepatitis and cirrhosis. Thus, measurement of serum CG as compared with serum TBA appears to be a sensitive liver test.     

Changes in the plethysmogram patterns of finger reactive hyperemia in nutritional (alcoholic) liver disease patients with increased serum monoamine oxidase activities

The activity levels of serum monoamine oxidase in nutritional (alcoholic) liver disease patients were elevated beyond the upper normal limit, while those of other subjects without pathologically accelerated fibrosis ranged within normal values. Capillaroscopic and plethysmographic studies on finger reactive hyperemia revealed that two characteristic changes in the blood microcirculation, which indicated an occurrence of increased fibritic process in the capillary wall and skin tissues, were observed only in the nutritional (alcoholic) liver disease patients with the elevated serum monoamine oxidase levels. These results suggested that nutritional (alcoholic) liver disease is not a disease restricted to liver tissues, but to a disorder creating pathological fibrosis in the whole-body.     

108例甲状腺功能障碍患者全血9种元素相关性研究

通过原子吸收光谱技术对68例治疗前甲亢病人(A组)，35例治疗后甲亢病人(B组)，5例甲减病人(C组)及74例正常人(D组）全血多种微量元素测定分析，结果显示：D组Cu与Al，Mg与Mn正相关；Al与Zn、Se负相关；A组Fe与Zn、Ca，Ca与TT4。Cu与Mn正相关，TT与Zn、Cr负相关。B组Fe与Zn、Cu、Mg，Zn与Cu、Mn、Mg与Ca、Se、Se与TT3正相关。     

A study on postsplenectomy changes of serum tuftsin level using a radioimmunoassay protocol

In this article, postsplenectomy changes of serum tuftsin level were studied on both human subjects and rabbits by using a self-established radioimmunoassay protocol. Antituftsin antibodies were raised in rabbits and roosters.¹²⁵I-tuftsin was prepared through an Iodogen method. The characteristics of the RIA were satisfactory with a detecting range of 0.5–100 ug/ml and the lowest detection limit of 400ng/ml. Serum tuftsin levels of splenectomized subjects were measured and compared with control groups. The serum tuftsin level from 30 postsplenectomy human beings was 406±179 ng/ml ( ± s) while that from a control group of 40 healthy blood donors was 557±256 ng/ml; the serum tuftsin level of 10 postsplenectomy rabbits was found to be 206±75 ng/ml while that of a control group of 10 normal rabbits was 318±96 ng/ml. The results showed that serum tuftsin level decreased markedly after splenectomy.     

Simultaneous Radioimmunoassay of Plasma Testosterone and Dihydrotestosterone

Abstract

This report describes the simultaneous radioimmunoassay of plasma testosterone (T) and 5a-dihydrotestosterone (DHT), using as binding reagent a highly specific antiserum which reacted significantly with T and DHT. Utilizing a one step celite microcolumn to separate T from DHT, we have measured these two androgens accurately in small aliquots of plasma from both male and female subjects, Plasma concentrations of T and DHT expressed as mean ± S.D. in ng/ml were respectively: 1) in male subjects: a. normal adult: 4.9 ± 1.6 (T) and 1.3 ± 0.8 (DHT); b. hypogonadal: 0.24 ± 0.08 (T) and 0.20 ± 0.10 (DHT); c. prepubertal: 0.14 ± 0.02 (T) and 0.05 ± 0.02 (DHT). 2) in female subjects: a. normally menstruating, irrespective of time of cycle: 0.22 ± 0.07 (T) and 0.34 ± 0.17 (DHT); b. postmenopausal: 0.29 ± 0.08 (T) and 0.15 ± 0.07 (DHT); c. ovariectomized: 0.24 ± 0.06 (T) and 0.09 ± 0.01 (DHT). When plasma levels of T and DHT were measured during a complete menstrual cycle, the levels of these steroids were found to be higher during the luteal than during the follicular phose. The ratio of plasma T:DHT concentrations was five times higher in normal adult male (3.1) than in menstruating female subjects (0.6).     

Iodine speciation in biological samples by capillary electrophoresis-inductively coupled plasma mass spectrometry.

A hyphenation of capillary electrophoresis (CE) to inductively coupled plasma mass spectrometry (ICP-MS) was employed for the speciation of iodine. The separation method used a buffer sandwich of phosphate (pH 2.3), NaOH, sodium dodecyl sulfate (SDS) and borate buffer (pH 8.3) for stacking, aiming at sufficient separation of iodide, iodate, thyroxine (T4) and triiodothyronine (T3). These four iodine species were separated within 15 min and subsequently detected during a pressure-driven detection step (baseline-separated) at 19.5, 29.1, 36.6 and 42.2 s. The detection limits were determined at 0.08 microg I/L (iodide), 0.3 microg I/L (iodate), 3.5 microg I/L (thyroxine) and 2.5 microg I/L (triiodothyronine). This method was applied on iodine speciation in human serum ("healthy" and after thyroid gland operation) and urine. The serum from the healthy person contained iodide (13 microg I/L), T4 (61 microg I/L) and T3 (7.5 microg I/L), whereas the serum from the thyroid-operated person lacked T3. As no "free" I-hormones are known in serum, the role of the thyroid hormone binding globulin (TBG) was investigated. We found that spiked T4 or T3 immediately bound to TBG. Investigations on human urine showed only a peak for iodide.     

Fundamental studies on thyroid stimulating hormone immunoradiometric assay kit (TSH RIABEAD II)

We have reported fundamental studies on the TSH immunoradiometric assay, using TSH RIABEAD II kit (Dainabot). The sensitivity of the assay was 0.03 mu IU/ml and its C.V. was 27.2%. Intra- and inter-assay C.V. were less than 5%. Dilution test and recovery test were good. Serum TSH level was 0.3-4.0 mu IU/ml in normal subjects, less than 0.03 mu IU/ml in untreated Graves' disease and subacute thyroiditis. Therefore, it was found that the clear difference exist in serum TSH levels between normal subjects and patients with untreated Graves' disease. There was a well correlation on the serum TSH levels between this method and TSH radioimmunoassay kit (Amerlex TSH, r = 0.983). Especially, the measurement of serum TSH levels, using immunoradiometric assay kit, was useful for the diagnosis of patients with Graves' disease.     

Comparison of Dechlorane Plus Concentrations in Sequential Blood Samples of Pregnant Women in Taizhou,China

To develop an appropriate sampling strategy to assess the intrauterine exposure to dechlorane plus (DP), we investigated DP levels in sequential maternal blood samples collected in three trimesters of pregnancy, respectively, from women living in Taizhou. The median concentration of DPs (sum of syn-DP and anti-DP) in all samples was 30.5 pg g⁻¹ wet-weight and 5.01 ng g⁻¹ lipid-adjusted weight, respectively. The trimester-related DP concentrations were consistently strongly correlated (p < 0.01), indicating that a single measurement of DP levels could represent intrauterine exposure without sampling from the same female repeatedly; however, the wet-weight levels significantly increased across trimesters (p < 0.05), while the lipid-adjusted levels did not significantly vary. Notably, whether lipid-adjusted weight or wet-weight levels, the variation extent of DP across trimesters was found to be less than 41%, and those for other persistent organic pollutants (POPs) reported in the literature were also limited to 100%. The limitation in variation extents indicated that, regardless of the time of blood collection during pregnancy and how the levels were expressed, a single measurement could be extended to screen for exposure risk if necessary. Our study provides different strategies for sampling the maternal blood to serve the requirement for assessment of in utero exposure to DP.     

Measurement of 24-hr whole-body retention of 99mTc-MDP with a thyroid uptake probe (author's transl)

A new method for measurement of 24-hr whole-body retention (WBR) of 99mTc-MDP, using a thyroid uptake probe was established and its clinical significance was evaluated in various bone diseases. (1) Reproducibility of 24-hr WBR in 9 patients was very good. Correlation coefficient was 0.997 and coefficient of variability was only 1.83%. (2) Radiochemical purity of 99mTc-MDP was 97.8 +/- 0.7% (n = 5), indicating no significant inter-lot variations. (3) 24-hr WBR of normal adult males (n = 5) was 30.0 +/- 4.9%, which was significantly elevated compared to the reported 99mTc-HEDP WBR of 19.2 +/- 1.7%. Whole-body retentions of chronic renal failure, metastatic bone disease and hyperthyroidism groups were significantly elevated compared to that of the normal group. However, WBR of steroid-induced osteoporotic group was significantly decreased. Based on these results, this thyroid uptake probe method was simple, reproducible and accurate to measure 24-hr WBR of 99mTc-MDP. Quantification of WBR of 99mTc-MDP was of great clinical value to diagnose metabolic bone disease and to follow-up metabolic and metastatic bone diseases.     

Quantitative determination of free and total bisphenol A in human urine using labeled BPA glucuronide and isotope dilution mass spectrometry

Bisphenol A (BPA) is a widely used industrial chemical in the manufacturing of polycarbonate plastic bottles, food and beverage can linings, thermal receipts, and dental sealants. Animal and human studies suggest that BPA may disrupt normal hormonal function and hence, potentially, have negative effects on the human health. While total BPA is frequently reported, it is recognized that free BPA is the biologically active form and is rarely reported in the literature. The objective of this study was to develop a sensitive and improved method for the measurement of free and total BPA in human urine. Use of a labeled conjugated BPA (bisphenol A-d6 β-d-glucuronide) allowed for the optimization of the enzymatic reaction and permitted an accurate determination of the conjugated BPA concentration in urine samples. In addition, a ¹³C₁₂-BPA internal standard was used to account for the analytical recoveries and performance of the isotope dilution method. Solid-phase extraction (SPE) combined with derivatization and analysis using a triple quadrupole GC-EI/MS/MS system achieved very low method detection limit of 0.027 ng/mL. BPA concentrations were measured in urine samples collected during the second and third trimesters of pregnancy in 36 Canadian women. Total maternal BPA concentrations in urine samples ranged from not detected to 9.40 ng/mL (median, 1.21 ng/mL), and free BPA concentrations ranged from not detected to 0.950 ng/mL (median, 0.185 ng/mL). Eighty-six percent of the women had detectable levels of conjugated BPA, whereas only 22 % had detectable levels of free BPA in their urine. BPA levels measured in this study agreed well with data reported internationally.     

Radioimmunoassay of Testosterone in Murine Plasma

Abstract

This report describes a radioimmunoassay method for measuring unconjugated plasma testosterone (T) and its application to the measurement of T in murine plasma. The parameters of assay reliability, sensitivity, and accuracy were established. Di-hydrotestosterone, the only steroid which showed significant cross-reaction with the testosterone antibody used, was not found in significant quantities in male mice of 19 inbred strains examined. This suggested that chromatographic separation of T from other plasma steroids was unnecessary for the measurement of T in males of this species. Biological levels of T in normal adult male mice of 19 inbred strains showed a range of 0.64 to 21.9 ng/ml plasma and a mean of 5.10 ng/ml. Castration reduced testosterone levels to about 4% of the normal level; castration plus adrenalectomy reduced testosterone to levels below the sensitivity of this method. These investigations demonstrated the validity and reliability of this method for measuring uncanjugated plasma T in male mice.     

Evaluation and comparison of high-sensitivity immunoradiometric assay kits for thyroid stimulating hormone

Fundamental and clinical characteristics of 3 kinds of high-sensitivity immunoradiometric assay (IRMA) kits for thyroid stimulating hormone (TSH). i.e., RIA BEADS II (kit A), TSH kit Daiichi II (kit B) and Ab tube TSH 'Eiken' (kit C) and one conventional radioimmunoassay (RIA) kit, i.e., TSH kit Daiichi (kit D), were studied. In the recovery test and the reproducibility test, there was no significant difference between the 4 kits. The sensitivities of kits A, B and C were much higher than that of kit D, and those IRMA kits were sensitive enough to distinguish hyperthyroidism from normal samples. For low concentrations of TSH (less than 5 microU/ml), the data from kits D, B, C and A tended to show higher values in that order. The correlation between the data measured by kits B and D, and the tendency of kit A toward lower values agreed well with other reports.     

A fundamental and clinical study of ferritin 'Eiken' radioimmunoassay kit

We have measured serum ferritin level using double antibody radioimmunoassay kit (Eiken ICL) and evaluated the characteristics of the kit and clinical usefulness. Satisfactory results were observed in standard curve, reproducibility, dilution and recovery test. In clinical evaluation, we have measured in normal subjects and patients with various diseases. The range in normal males and females were 13.0-158.7 ng/ml and 7.3-73.0 ng/ml, respectively. Serum ferritin level was elevated in patients with hepatoma, biliary cancer, lung cancer and other malignant diseases. Measurement of serum ferritin value would be useful in the monitoring of cancer patients.     

An enzyme-linked immunosorbent assay system for quantitative determination of calphobindin I, a new placental anticoagulant protein, and its application to various specimens

We developed a sandwich enzyme-linked immunosorbent assay (ELISA) system for calphobindin I (CPB-I), a new placental coagulation inhibitor, using two monoclonal antibodies. This ELISA system can detect CPB-I at concentrations of between 0.4 and 25 ng/ml in buffer and allow almost quantitative determination of it in human plasma. Using this ELISA system, CPB-I levels in many kinds of specimens were measured. Levels in the plasma and urine of women were as low as 10 ng/ml, and no significant differences were observed throughout the trimesters of pregnancy and during different stages of the menstrual cycle. Toxemic patients were slightly higher in CPB-I levels than normal pregnant women, and levels in body fluids such as the amniotic fluid, saliva, milk, ascites, and semen were higher than those in the plasma. The high levels of CPB-I were found, being in the order of micrograms/ml, in the ascites of carcinomatous peritonitis as well as seminal plasma. Measurements of the levels in ovarian follicular fluid samples at different stages of the menstrual cycle showed that those in the immature and atretic stages were higher than those in mature stages. CPB-I levels in many types of cultured human cells ranged from 0.023 to 10.30 micrograms/mg protein, and levels in cultured human lymphocytes were less than those in other types of cells measured. Little of this inhibitor was secreted into media from cultured human lymphocytes, and it was found in all measured tissues of Macacus irus at levels ranging from 0.232 to 1.557 micrograms/mg protein. From these results, it was suggested that CPB-I might be a ubiquitous protein in the body that has an important physiological role.     

Development of radioimmunoassay of intact laminin and its clinical evaluation as a tumor marker.

The concentration of laminin including laminin variants in serum samples was measured by a double-antibody radioimmunoassay using intact laminin. The mean level in 92 normal subjects (47 men and 45 women) was defined as 1 unit (U)/ml, and the cut-off value (2 S.D. above the mean) was 1.37 U/ml. Mean laminin level in 391 patients with various malignancies was 1.50 +/- 0.86 U/ml. Laminin levels were elevated in various cancer patients, and in 45.0% (176/391) the values exceeded the cut-off level; in patients with cancer of the stomach or pancreas, positivity rate exceeded 60%. Mean laminin concentration for 130 pregnant women (2.06 +/- 0.65 U/ml) was also significantly higher than that of normal controls, but concentrations for patients with various benign diseases were within a low range (0.55 +/- 0.29-1.10 +/- 0.29 U/ml). In the stomach or pancreas cancer patients, a positive correlation between laminin level and tumor progression or course of the disease was observed. These findings indicate that serum laminin level is potentially useful in the diagnosis and monitoring of certain cancers.     

Derivatization of Methylamine and Ethylamine Followed by LC and Fluorescence Detection for Measurement of Urinary Clearance

Monomethylamine (MMA) and ethylamine (EtA) accumulate in uremia. However, the urinary clearances of these compounds in normal humans and those with chronic kidney disease (CKD) have not been studied. In this work, an LC based fluorescence assay for MMA and EtA was developed. In applying the method, we found that some previous assays of urinary levels of MMA and EtA lead to overestimations. Normal subjects with average creatinine clearance of 82 ± 21 mL min^?1 demonstrate clearances for the two aliphatic amines less than the creatinine clearance, and in the range of urinary urea clearance. CKD subjects with an average creatinine clearance of 37 ± 12 mL min^?1 had modestly but significantly elevated serum levels of MMA and EtA above normals. Their fractional clearances were increased markedly compared to normals, mitigating the rise in their plasma levels. These findings imply enhanced secretion and/or reduced reabsorption of the amines with CKD.     

A new solid phase enzyme-immunoassay for total thyroxine (T4), triiodothyronine (T3) and thyroxine-binding globulin (TBG)

Conclusions When comparing the values determined by the radioimmunological and enzymeimmunological methods (Table 1), no significant differences were found in the Wilcoxon test for paired samples. Precision, specifity and accuracy of the EIA are comparable to those of the RIA. There are however important advantages of the EIA: 1) its extreme stability of antigen enzyme conjugates and 2) no further need of radiochemicals.

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Ein neuer solid-phase Enzymimmunoassay für Gesamt-Thyroxin (T₄), Triiodthyronin (T₃) und Thyroxin-bindendes Globulin (TBG)

     

Abnormal hepatobiliary clearance of 99mTc-N-(2,6-diethylphenylcarbamoylmethyl)iminodiacetic acid in the altered state of thyroid--by an analysis of deconvolution method

Hepatobiliary clearance of 99mTc-EHIDA was investigated in cases with altered thyroid function by deconvolution method. The results indicated that mean hepatic transit time of all control subjects revealed less than 10 minutes. On the other hand, mean hepatic transit time of cases with altered thyroid function revealed prolonged more than 13 minutes. Cases especially showing an elevated serum concentration of TSH compared with normal range (4.6 microU/ml) had a tendency of a high incidence of markedly prolonged mean hepatic transit time. These results suggest that thyroid hormone may influence on the hepatic metabolism of hepatobiliary radiopharmaceuticals. This phenomenon also could partly explain the cause of liver dysfunction seen in subjects with altered states.     

Zn、Mg、FT3、FT4在甲亢患者血清中的含量及相关性探讨

测定了３１例甲状腺机能亢进患者血清中Ｚｎ,Ｍｇ和Ｔ３,Ｔ４含量,并对其临床意义及相关性进行了探讨,同时作正常人对照,经统计学处理。结果表明：Ｍｇ明显降低（Ｐ〈０．０１）,Ｚｎ明显升高（Ｐ〈０．０１）,ＦＴ３,ＦＴ４明显升高（Ｐ〈０．０５）,但此两种元素含量与ＰＴ３,ＰＴ４水平没有等级相关性。