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Biomedical subjects

T H Cho

Publications and source records attributed to T H Cho.

15 recordsLinked to original sources
PubMed2000

Liquid chromatographic method for fumonisin B1 in sorghum syrup and corn-based breakfast cereals.

The fungus Fusarium verticillioides has been found on corn and sorghum, so it is possible that one or more of these toxins may be found in corn products such as breakfast cereals and syrup prepared from sorghum. Published methods when applied to syrups spiked with fumonisins gave low recoveries, less than 50%. A method was therefore developed which would be applicable to the syrup and breakfast cereals as well. Test samples were extracted with methanol-0.1 M sodium phosphate buffer (pH3) (1 + 1). The extract was diluted with water and applied to a 1 g C18 column. The column was washed with acetonitrile-water (2 + 8). Fumonisin B1 (FB1) was eluted with acetonitrile-trifluoroacetic acid (1000 + 1). The purified extract was evaporated and the toxin was derivatized with ophthaldialdehyde mercaptoethanol. The reaction mixture was resolved on a C18 liquid chromatographic column using acetonitrile-water-acetic acid (500 + 550 + 10.5) as the mobile phase at 37 degrees C, and FB1 measured with a fluorescence detector (excitation 335 nm, emission 440 nm). Recoveries of FB1 added to samples of sorghum syrup at levels ranging from 0.1 to 1.0 microgram/g were 94-132%. Recoveries of FB1 added to samples of breakfast cereal (corn flakes) at levels ranging from 0.2 to 1.6 micrograms/g were 96-100%. The method was applied to the analysis of 35 samples of sorghum syrup collected from 15 states in the US. One sample was found to contain FB1 at 0.12 microgram/g. A total of 32 samples of breakfast cereals collected by the Food and Drug Administration inspectors from grocery stores around the Kansas City area were analysed; no FB1 was found in the breakfast cereals (< 0.01 microgram/g). Results of this study indicated that FB1 possibly is not a problem in sorghum syrup and corn-based breakfast cereals in the US.

Biomarkers↗
PubMed1999

Magnetic resonance image-based cerebellar volumetry in healthy Korean adults.

The effects of age and gender on cerebellar size have not been established yet. To understand these effects, the area of cerebellar vermis and the volume of cerebellum were measured using serial magnetic resonance images of 124 Korean adults free of neurologic symptoms and signs. Cerebellar volume of male was significantly larger than that of female, although the size of vermis did not show significant gender difference. Correlation analysis revealed that cerebellar volume was not affected by aging. Regressional analysis demonstrated that female vermis had a tendency to shrink after age of 50, whereas male vermis and total cerebellar volume in both sexes were not altered with aging. The different response of vermis with aging and maintenance of cerebellum volume need to be more explored.

Adult↗
PubMed1999

Aloesin and arbutin inhibit tyrosinase activity in a synergistic manner via a different action mechanism.

In this study, we present evidence that cotreatment of aloesin and arbutin inhibits tyrosinase activity in a synergistic manner by acting through a different action mechanism. Aloesin or arbutin similarly inhibited enzyme activity of human- and mushroom-tyrosinases with an IC50 value of 0.1 or 0.04 mM, respectively. Lineweaver-Burk plots of the enzyme kinetics data showed that aloesin inhibited tyrosinase activity noncompetitively with a Ki value of 5.3 mM, whereas arbutin did it competitively (Maeda, 1996). We then examined whether cotreatment of these agents inhibits the tyrosinase activity in a synergistic manner. The results showed that 0.01 mM aloesin in the presence of 0.03 mM arbutin inhibited activity of mushroom by 80% of the control value and the reverse was also true. The inhibitory effects were calculated to be synergistic according to the Bürgi method. Taken together, we suggest that aloesin along with arbutin inhibits in synergy melanin production by combined mechanisms of noncompetitive and competitive inhibitions of tyrosinase activity.

Agaricales↗
PubMed1999

Administration of multiple cytokine genes with anti-tumor activity inhibits both tumor incidence and tumor growth.

The finding of reporter gene expression in muscle cells after intramuscular injection of a reporter gene containing DNA has suggested that injection of a certain gene in its naked form could induce an expression of the injected gene. The result proposed the concept, namely DNA or genetic vaccine technology, that injection of an antigen gene could induce a specific immune response against the antigen. Although the concept was initially applied to vaccination technology, the result also means that administration of cytokine genes with anti-tumor activity could exert their functions when they are applied as a naked form of DNA. To test the possibility, plasmid vector containing granulocyte macrophage-colony stimulation factor (GM-CSF) and interleukin-12 (IL-12) genes, which are known as one of the most potent anti-tumor cytokines, were constructed and injected into mice together with syngeneic tumor cells. When the cytokine gene containing plasmid was injected on the same day of tumor cell injection, a tumor mass developed in 4 out of 5 mice tested. Even among the 4 mice, the tumor mass of a mouse disappeared 2 weeks after tumor development. In addition, tumor generation was significantly delayed in cytokine gene injected mice and the average tumor size was about 51.5% that of vector control injected mice. These results suggested that tumor treatment through the injection of multiple cytokine genes with potent anti-tumor activity significantly inhibits tumor development and growth, and that the method could be considered as one of the tools for efficient tumor treatment.

Animals↗
PubMed1998

Structure and function of normal and transformed murine acyl-CoA binding proteins.

Acyl-CoA binding protein (ACBP) is a ubiquitous cytosolic protein found in high levels in tumorigenic cells. However, the molecular basis for the elevated levels of ACBP in malignant cells, ligand binding characteristics, and function in microsomal phospholipid synthesis have not been resolved. To address whether tumorigenic ACBP differs from the native protein, ACBP was purified from LM cells, a tumorigenic subline of mouse L-929 fibroblasts, and its primary structure was examined by delayed-extraction MALDI-linear TOF mass spectrometry. Proteolytic digestion and peptide sequence analysis confirmed that ACBP from LM cells was identical to native mouse ACBP (based on cDNA-derived amino acid sequence) with no amino acid substitutions, deletions, or posttranslational modifications. A fluorescent binding assay revealed that mouse ACBP bound cis-parinaroyl-CoA with high affinity, Kd 7.6 +/- 2.3 nM, at a single binding site. Furthermore, mouse ACBP enhanced microsomal phosphatidic acid formation from oleoyl-CoA 2.3-fold. Mouse ACBP also inhibited microsomal phospholipid acyl chain remodeling of choline-containing phospholipids, phosphatidylcholine and sphingomyelin, by 50 and 64%, respectively. These effects were specific compared to those of native rat liver or recombinant rat ACBP. Mouse and rat ACBPs differed by three amino acid substitutions at positions 4, 68, and 78. Although these small differences in amino acid sequence did not alter binding affinity for cis-parinaroyl-CoA, rat liver ACBP stimulated utilization of oleoyl-CoA 3.8-fold by microsomal glycerol-3-phosphate acyltransferase, significantly higher than that observed with mouse ACBP, but did not alter microsomal phospholipid acyl chain remodeling from oleoyl-CoA. In addition, these ACBPs protected oleoyl-CoA against hydrolysis. Finally, both mouse and rat ACBP shifted the incorporation of oleoyl-CoA from microsomal phospholipid acyl chain remodeling to phosphatidic acid biosynthesis. These data for the first time show a role for ACBP in stimulating microsomal phosphatidic acid biosynthesis and acyl chain remodeling in vitro. While ACBP from tumorigenic cells did not differ from normal, ACBPs from different murine species displayed subtle differences in their effects on microsomal phospholipid metabolism in vitro.

Acyl Coenzyme A↗
PubMed1998

Fatty acid binding protein isoforms: structure and function.

Although structural aspects of cytosolic fatty acid binding proteins (FABPs) in mammalian tissues are now well understood, significant advances regarding the physiological function(s) of these proteins have been slow in forthcoming. Part of the difficulty lies in the complexity of the multigene FABP family with nearly twenty identified members. Furthermore, isoelectric focusing and ion exchange chromatography operationally resolve many of the mammalian native FABPs into putative isoforms. However, a more classical biochemical definition of an isoform, i.e. proteins differing by a single amino acid, suggests that the operational definition is too broad. Because at least one putative heart H-FABP isoform, the mammary derived growth inhibitor, was an artifact (Specht et al. (1996) J. Biol. Chem. 271: 1943-49), the ensuing skepticism and confusion cast doubt on the existence of FABP isoforms in general. Yet, increasing data suggest that several FABPs, e.g. human intestinal I-FABP, bovine and mouse heart H-FABP, rabbit myelin P2 protein and bovine liver L-FABP may exist as true isoforms. In contrast, the rat liver L-FABP putative isoforms may actually be due either to bound ligand, post-translational S-thiolation and/or structural conformers. In any case, almost nothing is known regarding possible functions of either the true or putative isoforms in vitro or in vivo. The objective of this article is to critically evaluate which FABPs form biochemically defined or true isoforms versus FABPs that form additional forms, operationally defined as isoforms. In addition, recent developments in the molecular basis for FABP true isoform formation, the processes leading to additional operationally defined putative isoforms and insights into potential function(s) of this unusual aspect of FABP heterogeneity will be examined.

Animals↗
PubMed1998

Determination of aloesin in plasma by high-performance liquid chromatography as fluorescent 9-anthroyl derivative.

A sensitive high-performance liquid chromatographic (HPLC) method for the determination of aloesin in plasma was developed. After solid-phase extraction from plasma and derivatization of aloesin and compound AD-1, which was prepared from aloesin as a internal standard, with 9-anthroylnitrile in the presence of quinuclidine, the derivatives were separated on a inertsil ODS-3 column using acetonitrile/methanol/water (3:1:6) as a mobile phase, and detected fluorimetrically at 460 nm with excitation at 360 nm. The detection limit of aloesin was 3.2 ng/ml in plasma (S/N = 3).

Anthracenes↗
PubMed1997

Isoforms of rat liver fatty acid binding protein differ in structure and affinity for fatty acids and fatty acyl CoAs.

Although native rat liver fatty acid binding protein (L-FABP) is composed of isoforms differing in isoelectric point, their comparative structure and function are unknown. These properties of apo- and holo-L-FABP isoforms were resolved by circular dichroism, time-resolved fluorescence spectroscopy, and binding/displacement of fluorescent ligands. Both apo-isoforms had similar hydrodynamic radii of 18.5 A, but apo-isoform I had a greater alpha-helical content and exhibited a longer Tyr lifetime, indicative of secondary and tertiary structural differences from isoform II. Isoforms I and II both had two fatty acid or fatty acyl CoA binding sites. Ligand binding decreased the isoform hydrodynamic radii by 3-4 A and increased Tyr rotational motions in a more restricted range. Fatty acyl CoAs were more effective than fatty acids in altering the isoform structures. Scatchard analysis showed that both isoforms bound cis- parinaric acid with high affinity (Kd values 41 and 60 nM, respectively) and bound trans-parinaric acid with 2- and 7-fold, respectively, higher affinity than for cis-parinaric acid. In contrast, isoform I had higher affinity for cis- and trans-parinaroyl CoAs (Kd values of 33 and 14 nM) than did isoform II (Kd values of 110 and 97 nM), thereby resulting in biphasic plots of parinaroyl-CoA binding to native L-FABP. Finally, displacement studies indicated that each isoform displayed distinct specificities for fatty acid/fatty acyl CoA chain length and unsaturation. Thus, rat L-FABP isoforms differ markedly in both structure and ligand binding function.

Acyl Coenzyme A↗
PubMed1997

Lipid specificity and location of the sterol carrier protein-2 fatty acid-binding site: a fluorescence displacement and energy transfer study.

Although it was recently recognized that sterol carrier protein-2 (SCP-2) interacts with fatty acids, little is known regarding the specificity of SCP-2 for long-chain fatty acids or branched-chain fatty-acid-like molecules. Likewise the location of the fatty-acid binding site within SCP-2 is unresolved. A fluorescent cis-parinaric acid displacement assay was used to show that SCP-2 optimally interacted with 14-22 carbon chain lipidic molecules: polyunsaturated fatty acids > monounsaturated, saturated > branched-chain isoprenoids > branched-chain phytol-derived fatty acids. In contrast, the other major fatty-acid binding protein in liver, fatty-acid binding protein (L-FABP), displayed a much narrower carbon chain preference in general: polyunsaturated fatty acids > branched-chain phytol-derived fatty acids > 14- and 16-carbon saturated > branched-chain isoprenoids. However, both SCP-2 and L-FABP displayed a very similar unsaturated fatty-acid specificity profile. The presence and location of the SCP-2 lipid binding site were investigated by fluorescence energy transfer. The distance between the SCP-2 Trp50 and bound cis-parinaric acid was determined to be 40 A. Thus, the SCP-2 fatty-acid binding site appeared to be located on the opposite side of the SCP-2 Trp50. These findings not only contribute to our understanding of the SCP-2 ligand binding site but also provide evidence suggesting a potential role for SCP-2 and/or L-FABP in metabolism of branched-chain fatty acids and isoprenoids.

Animals↗
PubMed1996

Sterol carrier protein-2, a new fatty acyl coenzyme A-binding protein.

The ability of sterol carrier protein-2 (SCP-2) to interact with long chain fatty acyl-CoAs was examined. SCP-2 bound fluorescent fatty acyl-CoAs at a single site with high affinity. Kd values for cis- and trans-parinaroyl-CoA were 4.5 and 2.8 nM, respectively. Saturated 10-18-carbon and unsaturated 14-20-carbon fatty acyl-CoAs displaced SCP-2-bound fluorescent ligand. Oleoyl-CoA and oleic acid (but not coenzyme A) significantly altered SCP-2 Trp50 emission and anisotropy decay, thereby increasing SCP-2 rotational correlation time, SCP-2 hydrodynamic radius, and SCP-2 Trp50 remaining anisotropy up to 1.7-, 1.2-, and 1.3-fold, respectively. These changes were not accompanied by significant alterations in protein secondary structure as determined by circular dichroism. Finally, SCP-2 differentially altered the fluorescence emission and anisotropy decays of bound cis- and trans-parinaroyl-CoA. Both fluorescent fatty acyl-CoAs were located within a very ordered (limited cone angle of rotation) environment within SCP-2, as shown by a remaining anisotropy of 0.365 and 0.361 and a wobbling cone angle of 12 and 13 degrees , respectively. These anisotropy values were very close to those of such ligands in a propylene glass. However, the rotational relaxation times exhibited by SCP-2-bound cis- and trans-parinaroyl-CoA, 8.4-8.8 ns, were longer than those for the corresponding free fatty acid, 7.5-6.6 ns. These data show for the first time that SCP-2 is a fatty acyl-CoA-binding protein.

Acyl Coenzyme A↗
PubMed1992

Measurement of serum rubella antibody titers in Korean children, female adolescents, and pregnant women.

IgG antibody titers to rubella virus in serum of 322 nuchal cord blood, 1,473 children, and 1,103 reproductive and pregnant women were detected in Soonchunhyang University Hospital in Korea. Positive rate of IgG antibody to rubella virus was 86.3% (278/322) in nuchal cord blood, 68.0% (991/1,458) in children. The IgG antibody from the mother disappeared between the 7th to 11th month after birth, and then the positive rate increased as age increased. In the reproductive age group including pregnant women, positive rate of IgG antibody was 81.8% (894/1,103). Therefore, it was known that there was a possibility that 18.9% of this group would be attacked with rubella in Korea.

Adolescent↗
PubMed1991

Effects of Escherichia coli endotoxin on structure and permeability of myocardial capillaries.

A study was carried out to investigate changes in myocardial capillaries induced by endotoxin, in order to clarify the pathogenesis of myocardial damage in endotoxemia. Wistar rats were injected intraperitoneally with 100 mg/kg Escherichia coli lipopolysaccharide and then sacrificed at 1, 2, 3,4, 5, 6, 8, and 24 h after injection. The myocardium was observed by electron microscopy with histochemistry using horseradish peroxidase and immunocytochemistry for Na+, K(+)-ATPase/TPase. The earliest evident endothelial alterations were swelling, increased numbers of pinocytotic vesicles, and formation of cytoplasmic projections. Interstitial edema and focal detachment of the endothelial cells from the basement membrane occurred with time. Vascular permeability was increased after endotoxin injection. Activity of Na+, K(+)-ATPase was reduced on the plasma membrane of the endothelial cells. It is concluded that endotoxin induces structural and enzymatic changes in the myocardial capillary endothelium and an increase of capillary permeability.

Animals↗
PubMedDate not supplied

Determination of deoxynivalenol in white flour, whole wheat flour, and bran by solid-phase extraction/liquid chromatography: interlaboratory study.

A liquid chromatographic (LC) method for determining deoxynivalenol (DON) in white flour, whole wheat flour, and bran at or above the U.S. Food and Drug Administration advisory level of 1 microgram/g was evaluated by an interlaboratory study. Test samples of processed wheat (flour and bran) were extracted by blending with acetonitrile-water (84 + 16). Extracts were filtered and passed through a solid-phase extraction (SPE) column. The eluate was then chromatographed on a reversed-phase LC column with a water-methanol gradient. DON was measured at 220 nm. Naturally contaminated white flour, whole wheat flour, and bran samples and spiking solutions of DON to be added to the 3 commodities at 0.5, 1.0, and 2.0 micrograms/g were sent to 4 collaborators in Kansas, Louisiana, Missouri, and Washington states. Three collaborators completed the study. Average recoveries of DON from the 3 commodities spiked at 0.5, 1.0, and 2.0 micrograms/g were 94, 87, and 97%, respectively. Within-laboratory relative standard deviations for repeatability (RSDr) ranged from 3.1 to 21.7% and between-laboratory relative standard deviations for reproducibility (RSDR) ranged from 10.8 to 38.7%. On the basis of the results of this study, the SPE/LC method for DON in white flour, whole wheat flour, and bran was adopted as a peer-verified method by AOAC INTERNATIONAL.

Chromatography, Liquid↗