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Biomedical subjects

H Seo

Publications and source records attributed to H Seo.

At least 271 records · Page 15Linked to original sources

[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.

Evaluation Studies as Topic↗

[Hyponatremia with high plasma ANP level--report of two cases with emphasis on the pathophysiology of cerebral salt wasting].

Two cases of hyponatremia with intracranial lesions are reported with emphasis on diagnostic value of measurement of antidiuretic hormone (ADH) and atrial natriuretic polypeptide (ANP). Case 1. A 77-year-old female was transferred to our hospital for further care of vegetative state after subarachnoid bleeding on May 23, 1986. She was operated by neck clipping of rt-IC bifurcation aneurysm and lt-internal carotid-posterior communicating aneurysm at another hospital. On admission, computed tomography showed diffuse low density at bilateral thalamus and centrum semiovale. Biochemical analysis revealed hyponatremia (120 mEq/t) with increased natriuresis. Endocrinological date revealed normal plasma ADH and high plasma ANP levels. Patient was treated with infusion of 1% NaCl. Case 2. A 65-year-old male was admitted to our department because of gradual impairment of consciousness and generalized convulsion. Computed tomography showed small low density area at rt-thalamus and lt-cerebellar hemisphere. Biochemical date revealed severe hyponatremia (91 mEq/t) with normal plasma level of ADH and high plasma ANP. He was treated with infusion of 3% NaCl and hyponatremia was improved. The hyponatremia is frequently associated with intracranial disorders such as brain tumor, subarachnoid hemorrhage and head injury. Originally, hyponatremia with natriuresis was thought to be caused by salt wasting. This syndrome was defined as the inability to prevent salt loss in the urine due to undefined natriuretic factor in the brain. However, since 1957, because of introduction of concept of SIADH, it has generally become accepted that patients with natriuresis had SIADH. (ABSTRACT TRUNCATED AT 250 WORDS)

Aged↗

Seadragon VI: a 7-day dry saturation dive at 31 ATA. VI. Hyperbaria enhances renin but eliminates ADH responses to head-up tilt.

Three male subjects were passively tilted from a supine to a 90 degree head-up standing position on 2 d each at 1 and 31 ATA, then on 1 d of the postdive period. On each day the subjects were tilted once in the morning (0800-1000) and once in the evening (2000-2200). Before each tilt experiment, the subjects were first intravenously cannulated for blood sampling, then assumed the supine position. A blood sample was taken after 10 min in the supine position, and another sample was taken after 15 min of motionless, supported standing. The plasma was analyzed for antidiuretic hormone (ADH), plasma renin activity (PRA), plasma cortisol, and aldosterone. ADH, PRA, and cortisol were significantly increased by tilt, but the responses varied with time of day or atmospheric pressure. Cortisol increased only in the morning tilt (P less than 0.005) and was not affected by pressure. At 1 ATA, PRA was elevated in the morning tilt experiment (P less than 0.005) and not the evening tilt, but the overall response to tilt was greater at 31 ATA than at 1 ATA (P less than 0.005). The ADH response to tilt (P less than 0.025) was unaffected by time of day, but was eliminated at 31 ATA. The basal levels of ADH were also lower at 31 ATA (P less than 0.005). The mechanism of these responses remains unclear, but the eliminated postural stimulation of ADH may account for the eliminated circadian excretory pattern of the hormones. The altered responses to body fluid shifts possibly contribute to the increased aldosterone and decreased ADH frequently observed at hyperbaria.

Adult↗

Cloning and sequence analysis of cDNA for mouse prolactin.

The present study was undertaken to find out whether or not sexual dimorphism in biological activities and amino acid compositions of mouse prolactin might be due to heterogeneity in mRNA for mouse prolactin Cloned cDNAs for mouse prolactin were first isolated from a mouse pituitary cDNA library by hybridization with a rat prolactin cDNA. Then, one clone of about 140 positive clones obtained from 2000 transformants was subjected to nucleotide sequence analysis and verified to contain a nearly full length of cDNA sequence coding for mouse prolactin precursor. The deduced complete amino acid sequence indicates that the precursor molecule consists of 31 amino acids as the signal peptide and 197 amino acids of prolactin, in which two amino acids were found to be different from the amino acid sequence previously published elsewhere. S1 nuclease mapping analysis using male and female pituitary RNAs indicates that mouse preprolactin is encoded by two mRNAs in both sexes. The two mRNAs differ from each other based upon the deletion of three nucleotides in the coding region for the signal peptide determined by the nucleotide sequence analysis in other cDNA clones. In the present study, no sexual difference was revealed in murine prolactin mRNA.

Amino Acid Sequence↗

Effect of estrogen on prolactin mRNA in the rat pituitary. Analysis by in situ hybridization and immunohistochemistry.

The effect of estrogen on prolactin (PRL) synthesis at a single-cell level was studied by in situ hybridization and immunohistochemistry. Long-term estrogen treatment increased PRL-containing cells from 10-20% of total cell population to 80-90%, as revealed by immunohistochemistry. PRL mRNA containing cells also increased in a similar fashion. Moreover, cytoplasmic PRL mRNA expressed as the number of silver grains per cell increased 4- to 5-fold by estrogen. These results suggest that long-term estrogen treatment causes not only PRL cell proliferation but also an increase in PRL mRNA in a single cell.

Animals↗

Ontogenesis of pituitary prolactin in the human fetus.

To study the ontogenesis of PRL synthesis and secretion in the human fetus, PRL mRNA was measured in 12 pituitaries from fetuses of 16-27 weeks of gestation by hybridization of cytosol RNA with 125I-labeled single stranded complementary DNA (cDNA). Pituitary PRL content and serum PRL concentration were assessed by RIA. Pituitary weight increased with fetal age, ranging from 12 mg at 16 weeks of gestation to 41 mg at 27 weeks. The increased weight was due to an increase in cell number. PRL mRNA content did not change during weeks 16-20 of gestation. However, it increased rapidly after 21 weeks, reaching 11.5 times the 16-20 week value at 27 weeks. Pituitary PRL content was constant at a low level until 21 weeks, but thereafter it increased markedly. The increase was greater than that in PRL mRNA, and therefore, the PRL to PRL mRNA ratio was 10-fold greater at 27 weeks of gestation. Serum PRL concentrations also gradually increased after 21 weeks of gestation. These results indicate that marked increases in fetal pituitary PRL synthesis and release occur after 21 weeks of gestation.

Base Sequence↗

Comparative study of primates' transcortin: immunoreactivity and steroid-binding activity.

To investigate the phylogenic aspect of transcortin (corticosteroid-binding globulin, CBG), the immunoreactivity of transcortin with anti-human transcortin antiserum was studied in primates. The anti-human transcortin antibody was recognized by plasma proteins obtained from Catarrhini, taxonomically the most evolved monkey group. The immunoreactivity was not observed in plasma obtained from Platyrrhini and Prosimiae, classified as less evolved monkey groups than Catarrhini. Though comparison of immunoreactivity among different classes of Catarrhini was difficult because of non-parallelism of their displacement curves, displacement of 125I-labelled human transcortin from the antiserum by 1:10 and 1:100 diluted plasma was highest in human followed by Pongidae, Cercopithecoidea. The immunoreactivity of thyroxine-binding globulin (TBG) with anti-human TBG antiserum was also examined. The anti-human TBG antibody was only recognized in plasma from Pan (anthropoid ape) among Pongidae, highly evolved monkeys among Catarrhini. The existence of immunoreactive transcortin and TBG to respective human protein antibody in the highly evolved ape agreed well with the cladogenetic division of primate species delineated by Goodman and Moore (1971). Cortisol-binding activity of transcortin was detected in all monkeys except three, tafted capuchin monkey, night monkey and cotton-headed tamarin, which belong to Platyrrhini. The absence of cortisol-binding activity in these animals might be attributed to high levels of endogenous cortisol and low cortisol-binding capacity of transcortin. It is speculated that the structure of the immunoreactive site in transcortin could be modified by evolution without affecting the biologically important site, the site for cortisol binding.

Animals↗

Measurement of human prolactin messenger RNA in decidual tissues using complementary DNA probe cloned in M13mp9 bacteriophage.

A human prolactin (PRL) cDNA clone was digested with restriction enzyme Pst I and the resultant fragments were cloned into bacteriophage M13mp9. Single stranded recombinant DNAs having a coding strand of the PRL cDNA were selected by hybridization with 125I-labeled PRL mRNA obtained from human prolactinoma tissue. One of the single stranded recombinant DNAs was purified by agarose gel electrophoresis and labeled with 125I to a specific activity of 1.4 X 10(8) cpm per microgram of DNA. The probe could be successfully used in RNA dot hybridization. Analysis of poly (A) RNAs from prolactinoma, liver and placental tissues revealed that this probe was specific to PRL mRNA sequence. Hybridization of poly (A) RNA from decidual tissue to this probe revealed the presence of PRL mRNA sequence. However, PRL mRNA in decidua was at least 20,000 times less than that in pituitary prolactinoma.

Cloning, Molecular↗

Development of enzyme-linked immunospecific assay (ELISA) for the detection of monoclonal antibody: application to detection of monoclonal anti-TBG.

An enzyme-linked immunospecific assay (ELISA) was developed to screen monoclonal antibody to human thyroxine-binding globulin (TBG). The assay is based on the absorption of TBG from human pooled serum by rabbit polyclonal anti-TBG antibody coated on a microtiter plate and subsequent binding of monoclonal antibody to the absorbed antigen. Monoclonal antibody which binds TBG can be detected by peroxidase labeled anti-mouse IgG. From the examination with two different rabbit polyclonal anti-TBG antibodies, it was demonstrated that both purified IgG fraction and whole serum could be used as coating materials. It was not necessary to use purified TBG to react with anti-TBG antibody coated on the microtiter plate. When eight commercially available microtiter ELISA plates were tested in the assay, only 3 gave satisfactory results. The sensitivity of the assay was comparable with that of the conventional immunoprecipitation method using 125I-TBG and formalin fixed Staphylococcus aureus (Kowan strain) as an immunoabsorbent. The ELISA method could detect antibody activity in 0.032 microliter of medium obtained from a 3 day culture of confluent hybridoma cells. It is possible to store the antibody-antigen complexed microtiter plate for more than 2 weeks at 4 C. This makes possible rapid screening of monoclonal antibody.

Animals↗

Human transcortin synthesis by a cell-free translation of hepatic mRNA.

To evaluate the site of synthesis and to characterize the translated transcortin, poly (A)-containing RNA (mRNA) from human liver was translated in a cell-free system derived from rabbit reticulocyte lysate. The in vitro synthesized product was identified as transcortin by immuno-precipitation with its specific antiserum. This translated transcortin could be displaced from the antibody by unlabeled purified transcortin obtained from plasma. Furthermore, when the translation mixture was applied to a cortisol-Sepharose column, the translated transcortin was bound to the matrix in a specific manner, indicating that this product binds to cortisol. The molecular weight of the translated transcortin was estimated to be 45,700 by its mobility in sodium dodecyl sulfate polyacrylamide gel electrophoresis, while that of plasma transcortin was 53,800. The difference in molecular weight between the translated transcortin and plasma transcortin was probably due to the presence of pre-sequence (signal peptide) in addition to the absence of carbohydrate moiety in the former. In conclusion, human liver mRNA directed the synthesis of transcortin, and the translated transcortin binds to cortisol in spite of the absence of carbohydrate moiety.

Animals↗

Mechanism of interference between influenza A/WSN and B/Kanagawa viruses.

Simultaneous infection of MDCK cells with influenza viruses A/WSN and B/Kanagawa resulted in mutual interference with virus protein synthesis and in significant suppression of A/WSN growth. When infection by one virus preceded the other by 1 or 2 h, growth of the superinfecting virus was selectively inhibited at the level of transcription. Interference by the pre-infecting virus was strongly dependent on the expression of the viral genome but not on haemagglutinin activity. When the replication of both virus types was restricted to primary transcription by cycloheximide, the only translation products following removal of the drug were those of the preinfecting virus. This result was not affected by blocking secondary transcription by actinomycin D. These findings suggest that intertypic interference occurs at the level of primary transcription. This concept was supported further by the observation that a ts mutant of A/WSN (ts-65) with a defect in primary transcription interfered only with superinfection by B/Kanagawa at the permissive temperature.

Animals↗

Improvement of the oral bioavailability of digitalis glycosides by cyclodextrin complexation.

Inclusion complexes of the digitalis glycosides digitoxin, digoxin, and methyl digoxin with three cyclodextrins (alpha-, beta-, gamma-homologues) in water and in the solid state were studied by a solubility method, IR and 1H-NMR spectroscopy, and X-ray diffractometry. Solid complexes (in a molar ratio of 1:4) of the digitalis glycosides with gamma-cyclodextrin were prepared and their in vivo absorption examined. The rapidly dissolving form of the gamma-cyclodextrin complex significantly increased plasma levels of digoxin (approximately 5.4-fold) after oral administration to dogs.

Animals↗

Effect of high salt treatment on influenza B viral protein synthesis in MDCK cells.

Based on the information that high salt inhibits the initiation of cellular mRNA translation which depends on the function of the 5'-terminal structure of mRNA, we compared the effect of high salt on translation of host cellular mRNAs and influenza viral mRNAs, both of which are of 5'-terminal structure. Brief exposure of influenza B virus-infected MDCK cells to high salt medium resulted in a dose-dependent inhibition of viral polypeptide synthesis as well as of cellular polypeptide synthesis, but it had less effect on synthesis of viral polypeptides, particularly nonstructural protein (NS). Under these conditions the Na+ content of the infected cells was significantly increased. A similar salt effect on in vitro translation of viral and cellular mRNAs extracted from infected cells was also observed. There was no significant difference in sensitivity to hypertonic block of in vivo translation of influenza viral mRNAs and vesicular stomatitis virus mRNAs, the latter of which possess a virus-directed structure at the 5'-terminus.

Animals↗