Search PubMed⌕ Search

Biomedical subjects

K W Cho

Publications and source records attributed to K W Cho.

At least 109 records · Page 6Linked to original sources
PubMed1992

Frog lymph heart synthesizes and stores immunoreactive atrial natriuretic peptide.

The presence of immunoreactive atrial natriuretic peptide (irANP) and ANP gene expression in the frog lymph heart was examined by a radioimmunoassay (RIA) combined with HPLC and by Northern blot hybridization of total RNA. Serial dilution curve of the lymph heart extract was paralleled with the RIA standard curve. The lymph heart contained 153.32 +/- 35.80 pg of irANP/mg of wet tissue. The major form of irANP in the frog lymph heart was high molecular weight on reverse-phase and gel permeation high performance liquid chromatography as in the frog atria and ventricles. The frog lymph heart, as well as frog atria and ventricles, was shown to express mRNA coding for ANP. Dense core secretory granules similar to those observed in the mammalian atria were also found in the frog lymph heart. The presence of irANP and the expression of ANP gene in the frog lymph heart suggest that the lymph heart may participate in the regulation of homeostasis of lymph circulation and blood volume change through the synthesis and release of ANP.

Animals↗
PubMed1992

Ovarian atrial natriuretic peptide during the rat estrous cycle.

The changes in ovarian levels of immunoreactive atrial natriuretic peptide (irANP) and arginine vasopressin (irAVP) were observed during the estrous cycle of rat. We also demonstrated the synthesis of ovarian ANP. In adult 4-day cycling rats, ovarian level of irANP was found to be the highest on proestrus and was to be the lowest on diestrus. Ovarian irANP level inversely correlated with ovarian level of irAVP. On reverse-phase HPLC, two distinct peaks of ovarian irANP, high and low molecular weight forms, existed in the each stage of the estrous cycle. However, no significant changes in plasma and atrial concentrations of ANP were observed during the cycle. The rat ovary contained mRNA coding for ANP. These data showing the synchronized cyclic change of ovarian irANP and irAVP with the estrous cycle suggest that the ovary locally synthesizes ANP and ovarian ANP may play regulatory roles on the follicular fluid dynamics.

Analysis of Variance↗
PubMed1991

Molecular nature of Spemann's organizer: the role of the Xenopus homeobox gene goosecoid.

This study analyzes the function of the homeobox gene goosecoid in Xenopus development. First, we find that goosecoid mRNA distribution closely mimics the expected localization of organizer tissue in normal embryos as well as in those treated with LiCl and UV light. Second, goosecoid mRNA accumulation is induced by activin, even in the absence of protein synthesis. It is not affected by bFGF and is repressed by retinoic acid. Lastly, microinjection of goosecoid mRNA into the ventral side of Xenopus embryos, where goosecoid is normally absent, leads to the formation of an additional complete body axis, including head structures and abundant notochordal tissue. The results suggest that the goosecoid homeodomain protein plays a central role in executing Spemann's organizer phenomenon.

Activins↗
PubMed1991

Organizer-specific homeobox genes in Xenopus laevis embryos.

The dorsal blastopore lip of the early Xenopus laevis gastrula can organize a complete secondary body axis when transplanted to another embryo. A search for potential gene regulatory components specifically expressed in the organizer was undertaken that resulted in the identification of four types of complementary DNAs from homeobox-containing genes that fulfill this criterion. The most abundant of these encodes a DNA-binding specificity similar to that of the Drosophila melanogaster anterior morphogen bicoid. The other three are also homologous to developmentally significant Drosophila genes. These four genes may participate in the regulation of the developmental potential of the organizer.

Amino Acid Sequence↗
PubMed1991

Overexpression of a homeodomain protein confers axis-forming activity to uncommitted Xenopus embryonic cells.

The anteroposterior character of mesoderm induced by a peptide growth factor (XTC-MIF) was tested by transplantation into host Xenopus gastrulae. Both retinoic acid and a homeodomain protein were able to override the anteriorizing effect of the growth factor. Microinjection of a posteriorly expressed homeobox mRNA can respecify anteroposterior identity, transforming head mesoderm into tail-inducing mesoderm. Unexpectedly, overexpression of XIHbox 6 protein in the transplanted cells, without addition of growth factors, caused the formation of tail-like structures. The cells overexpressing XIHbox 6 were able to recruit cells from the host into the secondary axis. The results suggest that vertebrate homeodomain proteins are part of the biochemical pathway leading to the generation of the body axis.

Activins↗
PubMed1991

Identification of immunoreactive atrial natriuretic peptide in bulbus arteriosus of freshwater teleostean fish.

1. The presence and partial characterization of immunoreactive atrial natriuretic peptide (ir-ANP) in the bulbus arteriosus were investigated using high performance liquid chromatography (HPLC) and radioimmunoassay (RIA) in several freshwater teleostean fish (catfish, silver crusian carp, snakehead, and rice eel). 2. The level of ir-ANP in the bulbus arteriosus was comparable with that in the atria at about 5-15 pg/mg, 6-13 pg/mg wet wt, respectively, but was 2-4-fold higher than that in the ventricles in all species observed. 3. The HPLC profile of ir-ANP showed two main peaks corresponding to low and high mol. wt rat ANPs. 4. Our results provide evidence for the presence of ir-ANP with high as well as low mol. wts in the bulbus arteriosus of freshwater teleostean fish.

Animals↗
PubMed1991

Role of membrane sulfhydryl groups in stimulation of renin secretion by sulfhydryl reagents.

The present study was designed to address the reactivity and accessibility of the particular class of sulfhydryl groups involved in the regulatory process of renin secretion. Both mercurial (such as P-chloromercuriphenyl sulfonate [PCMPS] and non-mercurial sulfhydryl reagents (for example, 6,6-dithiodinicotinic acid [DTDN]), which very slowly penetrate the cell membrane of intact cells, stimulated renin secretion. The membrane permeant sulfhydryl reagent N-ethylmaleimide had no effect on renin secretion but its membrane impermeant derivative, stilbene maleimide, strongly stimulated secretion. Furthermore, disulfide reducing agents such as dithiothreitol (DTT) had no effect on renin secretion at low concentrations, but strongly inhibited it at high concentrations. Several reagents which are known to primarily deplete cellular reduced glutathione were without effect on renin secretion. The stimulation of renin secretion by PCMPS was rapid in onset, and prevented and reversed by DTT and L-cysteine. Furthermore, the maximal stimulatory effect of PCMPS was not additive to that by diuretics with sulfhydryl reactivity (such as, ethacrynic acid and mersalyl). The stimulatory effect of PCMPS was not affected by diuretics which lack sulfhydryl reactivity (such as, bumetanide and furosemide). These results suggest that sulfhydryl reagents of both with and without diuretic activity stimulate renin secretion by reacting with specific class of sulfhydryl groups which are readily accessible from the extracellular compartment. In addition, these results provide further support the possibility that a sulfhydryl-disulfide interchange in the membrane may play a regulatory role in the renin secretory process.

4-Chloromercuribenzenesulfonate↗
PubMed1991

Cloning and nucleotide sequences of lux genes and characterization of luciferase of Xenorhabdus luminescens from a human wound.

Xenorhabdus luminescens HW is the only known luminous bacterium isolated from a human (wound) source. A recombinant plasmid was constructed that contained the X. luminescens HW luxA and luxB genes, encoding the luciferase alpha and beta subunits, respectively, as well as luxC, luxD, and a portion of luxE. The nucleotide sequences of these lux genes, organized in the order luxCDABE, were determined, and overexpression of the cloned luciferase genes was achieved in Escherichia coli host cells. The cloned luciferase was indistinguishable from the wild-type enzyme in its in vitro bioluminescence kinetic properties. Contrary to an earlier report, our findings indicate that neither the specific activity nor the size of the alpha (362 amino acid residues, Mr 41,389) and beta (324 amino acid residues, Mr 37,112) subunits of the X. luminescens HW luciferase was unusual among known luminous bacterial systems. Significant sequence homologies of the alpha and beta subunits of the X. luminescens HW luciferase with those of other luminous bacteria were observed. However, the X. luminescens HW luciferase was unusual in the high stability of the 4a-hydroperoxyflavin intermediate and its sensitivity to aldehyde substrate inhibition.

Amino Acid Sequence↗
PubMed1991

Atrial pressure, distension, and pacing frequency in ANP secretion in isolated perfused rabbit atria.

It has been suggested in this laboratory that the principal stimulus for the secretion of atrial natriuretic peptide (ANP) is the reduction of atrial distension and that the secretion of ANP is dependent on both atrial reduction volume and reduction frequency. To investigate the relationship among the changes in atrial pressure, distension, pacing frequency, and ANP secretion, we performed a series of experiments in the isolated perfused rabbit atria. Increase in atrial pressure without changes in transmural pressure and thus without volume changes did not raise immunoreactive ANP (irANP) secretion. Atrial distension without changes in intracavitary atrial pressure increased irANP secretion with the reduction. Electrical stimulation with atrial distension resulted in an increase in irANP secretion in proportion to pacing frequency. Incremental response of irANP secretion to electrical stimulation was accentuated by increasing atrial distension. Neither atrial pacing without distension nor distension without pacing raised irANP secretion. These results suggest that the direct and principal stimulus for irANP secretion in response to atrial pacing and distension is the length shortening of atrial myocytes and that the incremental response of irANP secretion to increasing pacing frequency is the result of an increase in frequency of the length shortening of atrial myocytes.

Animals↗
PubMed1991

Gradient fields and homeobox genes.

We review here old experiments that defined the existence of morphogenetic gradient fields in vertebrate embryos. The rather abstract idea of cell fields of organ-forming potential has become less popular among modern developmental and molecular biologists. Results obtained with antibodies directed against homeodomain proteins suggest that gradient fields may indeed be visualized at the level of individual regulatory molecules in vertebrate embryos.

Animals↗
PubMed1991

Preferential distribution of active RNA polymerase II molecules in the nuclear periphery.

We have combined immunogold labeling with the Miller spreading technique in order to localize proteins at the electron microscope (EM) level in whole mount nuclei from mouse and human fibroblasts. Anti-histone H1 antibody labels nuclei uniformly, indicating that the nuclear interior is accessible to both antibodies and gold conjugates. Anti-topoisomerase I antibody labels nucleoli intensely, in agreement with previous immunofluorescent and biochemical data. Two different antibodies against the large subunit of RNA polymerase II (pol II) show preferential labeling of the nuclear periphery, as do antibodies against lamin, a known peripheral nuclear protein. Treatment of cells with alpha-amanitin results in loss of virtually all RNA polymerase II staining, supporting the specificity of labeling. Finally, when nuclei are incubated in the presence of biotin-UTP (bio-UTP) under run-off transcription conditions, incorporation is preferentially located at the nuclear periphery. These results support the conclusions that transcriptionally active pol II molecules are non-uniformly distributed in fibroblast nuclei, and that their differential distribution mirrors that of total pol II.

Amanitins↗
PubMed1990

Sequential mechanism of atrial natriuretic peptide secretion in isolated perfused rabbit atria.

It is well known that the secretion of atrial natriuretic peptide (ANP) is dependent on the atrial stretch. It has been claimed in this laboratory that the secretion of ANP occurs with a reduction in atrial distension. It was shown in the present experiment that the secretion of immunoreactive (ir) ANP occurs coincidently with a translocation of extracellular space marker (3-H)-inulin in the isolated perfused rabbit atria. Translocation of extracellular space fluid was observed with a reduction in atrial distension. The secretion of irANP into the atrial lumen occurs less than 15 sec of the reduction in atrial distension. It is therefore suggested that the incremental response of irANP secretion to the reduction in atrial distension is a sequential mechanism of ANP secretion, in which first is the release of ANP from the atrial myocytes into the extracellular space and then second is the translocation of ANP with extracellular space fluid into the atrial lumen with a reduction in atrial distension.

Animals↗
PubMed1990

Elicitation of an oxidase activity in bacterial luciferase by site-directed mutation of a noncatalytic residue.

Flavin-dependent external monooxygenases and oxidases could catalyze the same flavin oxidation reaction involving distinct mechanisms. To gain insights into enzyme structure-function relationship, site-directed mutagenesis was carried out for Vibrio harveyi luciferase, a monooxygenase. The substitution of the alpha subunit cysteine 106 by alanine shows unambiguously that the alphaCys106 is not essential to catalysis. The corresponding substitution by valine resulted in a substantial reduction of the bioluminescence activity correlatable with the induction of a new flavin oxidation activity typical for oxidases. These findings indicate that mutation of a single noncatalytic residue at the active center of a flavoenzyme could transform one enzyme type to another, thus highlighting the subtlety of enzyme active site structure in relation to catalysis and the versatility of enzyme evolution.

Catalysis↗
PubMed1990

Presence of immunoreactive atrial natriuretic peptides in pericardial fluid of human subjects with congenital heart diseases.

The epicardial release of immunoreactive atrial natriuretic peptides (ir-ANPs) in inside-out perfused rabbit atria has been reported. In order to determine the presence of ir-ANPs in pericardial fluid and to evaluate their biochemical characteristics, we measured the concentration of ir-ANPs in pericardial fluid obtained from the patients with congenital heart diseases during open heart surgery. Serial dilution curves made with the extrats of pericardial fluid using Sep-Pak C18 cartridges were parallel with standard curve. The concentration of ir-ANPs in pericardial fluid was significantly lower than the corresponding plasma concentration. On gel permeation and reverse-phase high performance liquid chromatography, the ir-ANPs in pericardial fluid, plasma and atrial appendage showed both high and low molecular weights. The major peak of ir-ANPs in plasma was observed at the corresponding fraction to the alpha-human ANP and considerable amount of high molecular weight form of ir-ANPs was observed in pericardial fluid. However, the major peak of ir-ANPs in atrial appendage was observed at the corresponding fraction to the rat pro-ANP. The data suggest that ir-ANPs exist both high and low molecular weight forms in pericardial fluid.

Atrial Natriuretic Factor↗
PubMed1990

A time-dependent bacterial bioluminescence emission spectrum in an in vitro single turnover system: energy transfer alone cannot account for the yellow emission of Vibrio fischeri Y-1.

Yellow fluorescent protein (YFP), which has a bound FMN, was isolated from the marine bacterium Vibrio fischeri strain Y-1b. Its presence in a luciferase [alkanal monooxygenase (FMN-linked); alkanal, reduced-FMN:oxygen oxidoreductase (1-hydroxylating, luminescing), EC 1.14.14.3] reaction mixture causes a striking color change, and an increase in bioluminescence intensity, as well as a faster rate of intensity decay, so that the quantum yield is not changed. The emission spectrum shows two distinct color bands, one at 490 nm attributed to the unaltered emission of the luciferase system, the other peaking in the yellow around 540 nm due to YFP emission. The kinetics of the two color bands differ, so the spectrum changes with time. The yellow emission reaches its initial maximum intensity later than the blue, and then both blue and yellow emissions decay exponentially with nearly the same pseudo-first-order rate constants, linearly dependent on [YFP] (from 0.01 sec-1 with no YFP to a maximum of approximately 0.1 sec-1 at 4 degrees C) but exhibiting a saturation behavior. The data can be interpreted by assuming the interaction of YFP with the peroxyhemiacetal intermediate in the luciferase reaction to form an unstable new complex whose breakdown gives the yellow emitter in its excited state. This simple model fits well the data at [YFP] less than 15 microM. The results indicate that a single primary excited state cannot be responsible for the blue and the yellow emissions.

Bacterial Proteins↗
PubMed1990

Differential activation of Xenopus homeo box genes by mesoderm-inducing growth factors and retinoic acid.

What is the nature of positional information during embryogenesis? By using Xenopus homeo box genes as anteroposterior (A-P) markers, we confirm the findings of others that mesoderm-inducing growth factors and retinoic acid (RA) can provide positional information along the axis of the body. Xenopus tissue culture-mesoderm-inducing factor (XTC-MIF) selectively activates an anteriorly expressed homeo box gene (XlHbox 1), while basic fibroblast growth factor (bFGF) activates selectively a posteriorly expressed homeo box gene (XlHbox 6). RA activates expression of the posterior gene XlHbox 6, but not of XlHbox 1. This activation, however, requires exposure to growth factors. The data suggest that growth factors and RA may cooperate with each other to provide positional information in vertebrates.

Animals↗
PubMed1989

Interference with function of a homeobox gene in Xenopus embryos produces malformations of the anterior spinal cord.

XIHbox 1 is expressed in a narrow band across the cervical region of Xenopus embryos. The gene produces two related proteins: "long" and "short" XIHbox 1 homeodomain proteins. Injection of antibodies to the long XIHbox 1 protein into 1-cell embryos caused a phenotype in which the anterior spinal cord was morphologically transformed into a hindbrain-like structure. This alteration was restricted to the region normally expressing long XIHbox 1 protein. Injection of long protein mRNA disrupted segmentation and tissue organization without inhibiting cell proliferation. Injection of short protein mRNA into 1-cell embryos produced spinal cord malformations similar, but not identical, to those caused by the antibodies, suggesting antagonistic roles for long and short XIHbox 1 proteins. We immunostained tadpoles carrying extended hindbrains for N-CAM and consistently found defective organization of spinal nerves over the affected region.

Animals↗