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Biomedical subjects

H K Lee

Publications and source records attributed to H K Lee.

At least 73 records · Page 4Linked to original sources

Application of static liquid-phase microextraction to the analysis of organochlorine pesticides in water.

Static liquid-phase microextraction, with subsequent analysis by gas chromatography-electron-capture detection, has been applied to extract eight organochlorine pesticides from water. A conventional microsyringe was used to extract analytes from water samples over a concentration range of 0.05-100 microg/l. Factors relevant to the extraction process were investigated. The sensitivity of the method was enhanced with agitation, and increasing the extraction temperature, of the sample solution. Concentration factors of >50-fold were easily achieved within 25 min of extraction. The analytical data exhibited a relative standard deviation (RSD) range of 3.2% (lindane) to 10.7% (methoxychlor) for the eight pesticides; most RSD values were under 7%. Water samples collected from a reservoir, and from tap water in a chemical laboratory were analyzed using the procedure.

Chromatography, Liquid↗

Antitumor agents. Part 204: synthesis and biological evaluation of substituted 2-aryl quinazolinones.

A series of 2',3',4',6,7-substituted 2-aryl quinazolinones were synthesized and evaluated for biological activity. Among them, 17 displayed significant growth inhibitory action against a panel of tumor cell lines. Compound 17 was also a potent inhibitor of tubulin polymerization. Compounds 8-10 displayed selective activity against P-gp-expressing epidermoid carcinoma of the asopharynx.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

Software and internet resources for capillary electrophoresis and micellar electrokinetic capillary chromatography.

The purpose of the contribution is to show how software and the internet have changed the way in which we carry out research, and what additional possibilities these new resources and tools provide. The internet and e-mail broaden our horizon for cooperation. There are no borders for information exchange: our library is a virtual one, electronic databases are at our fingertips. E-mail discussion groups provide an electronic community of CE users. Such forums have provided a basis for worldwide scientific cooperation amongst scientists; the present contribution is only one of several examples. Estimation software provides us with estimates of component properties in those cases where this information is not available from literature or from experiments (an estimated value is better than no value). Several examples will illustrate the use of estimation software in capillary zone electrophoresis and micellar electrokinetic capillary chromatography. Simulation software presents visualization of experimental results to be expected, both as a training tool and as the first step in method development. Other simulations yield valuable insights into phenomena that are not readily accessible experimentally for reasons of size or time-scale.

Chromatography, Micellar Electrokinetic Capillary↗

Inhibition of cytokine-induced vascular cell adhesion molecule-1 expression; possible mechanism for anti-atherogenic effect of Agastache rugosa.

Adhesion molecules such as vascular cell adhesion molecule-1 (VCAM-1) play an important role during the early stages of atherogenesis. Agastache rugosa has an anti-atherogenic effect in low density lipoprotein receptor -/- mice. Moreover, A. rugosa reduced macrophage infiltration and VCAM-1 expression has been localized in aortic endothelium that overlies early foam cell lesions. This study ascertained that tilianin (100 microM), a major component of A. rugosa, inhibits the tumor necrotic factor-alpha (TNF-alpha)-induced expression of VCAM-1 by 74% in cultured human umbilical vein endothelial cells (HUVECs). Also, tilianin (100 microM) reduced TNF-alpha-induced activation of nuclear factor-kappaB in HUVECs.

Animals↗

Platelet-activating factor-induced early activation of NF-kappa B plays a crucial role for organ clearance of Candida albicans.

In this study, we have investigated the mechanisms underlying organ susceptibility to candida infection. Infection of BALB/c mice with Candida albicans led to both an early (1-8 h) and late (24-48 h) activation of NF-kappaB in the organs resistant to C. albicans, including the lung and spleen. In susceptible organs such as the kidneys, early activation of NF-kappaB was not observed. The kinetics of TNF-alpha mRNA expression paralleled those of NF-kappaB activation in all organs examined. Blocking the effects of endogenous platelet-activating factor (PAF) by pretreatment with the PAF antagonist BN50739 or antioxidants significantly reduced the early activity of NF-kappaB and TNF-alpha mRNA expression, and increased the recovery of C. albicans in the lung and spleen. Importantly, administration of PAF 5 min prior to the infection resulted in the appearance of early activities of NF-kappaB and TNF-alpha mRNA expression, followed by a nearly complete clearance of the organisms in the kidneys. Pretreatment with anti-TNF-alpha Ab resulted in an enhanced susceptibility to C. albicans, and the PAF-mediated resistance was abrogated by anti-TNF-alpha in all organs examined. These data indicated that endogenously produced PAF in response to C. albicans is a key molecule involved in the early activation of NF-kappaB, which, in turn, renders the organ resistant to the fungus by promoting the production of anti-candidal proinflammatory cytokines such as TNF-alpha. Susceptible organs, including the kidneys, lack the capacity to generate a sufficient PAF-induced early NF-kappaB response.

Adjuvants, Immunologic↗

High-performance chiral separation of fourteen triazole fungicides by sulfated beta-cyclodextrin-mediated capillary electrophoresis.

In this paper, sulfated beta-cyclodextrin-mediated capillary electrophoresis (CE) is evaluated as a new approach for the chiral separation of triazole-type fungicides. The 14 fungicides investigated were bitertanol, cyproconazole, difenoconazole, diniconazole, flutriafol, hexaconazole, myclobutanil, paclobutrazol, penconazole, propiconazole, tebuconazole, tetraconazole, triadimefon and triadimenol. Under the optimal conditions, excellent enantioseparation was achieved for all the 14 fungicides, including those fungicides containing two chiral centers. To our knowledge, this is the only system to date that offers outstanding enantiodiscrimination towards all triazole-type fungicides. The impact of the molecular structures of the triazole compounds on their migration behavior was studied. Similar to other chemical systems involving host-guest complexation, the interaction between sulfated beta-cyclodextrin and the triazole compounds was found to be affected by a variety of factors, including electrostatic force, hydrogen bonding, steric effect and hydrophobicity. These factors, coupled with the countercurrent electroosmotic flow (EOF), were believed to be the major forces behind the exceptional chiral selectivity.

Cyclodextrins↗

Somatic mutations of TRAIL-receptor 1 and TRAIL-receptor 2 genes in non-Hodgkin's lymphoma.

Tumor necrosis factor-related apoptosis-inducing ligand-receptor 1 (TRAIL-R1) and tumor necrosis factor-related apoptosis-inducing ligand-receptor 2 (TRAIL-R2) are cell-surface receptors involved in tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced cell-death signaling. TRAIL-R1 and TRAIL-R2 genes have recently been mapped to chromosome 8p21-22, which is a frequent site of allelic deletions in many types of human tumors, including non-Hodgkin's lymphoma (NHL). Because TRAIL/TRAIL receptor system plays an important role in lymphocyte homeostasis, we hypothesized that the mutations of TRAIL-R1 and TRAIL-R2 may be involved in the development of NHL and that such mutations may be responsible for the allelic losses of 8p21-22 in NHL. In this study, we analysed the entire coding region of TRAIL-R2 gene and the death domain region of TRAIL-R1 gene for the detection of the somatic mutations in a series of 117 human NHLs using polymerase chain reaction (PCR)-based single strand conformation polymorphism (SSCP) analysis. Overall, eight tumors (6.8%) were found to have two TRAIL-R1 gene mutations or six TRAIL-R2 gene mutations. Interestingly, of the eight mutations, six missense mutations (two TRAIL-R1 and four TRAIL-R2) were detected in the death domains and one nonsense mutation of TRAIL-R2 was detected just before the death domain. Our data suggest that somatic mutations of TRAIL-R1 and TRAIL-R2 genes may play a role in the pathogenesis of some NHLs and that TRAIL-R1 and TRAIL-R2 genes might be the relevant genes to the frequent loss of chromosome 8p21-22 in human NHL.

Humans↗

Somatic mutations in the death domain of the Fas (Apo-1/CD95) gene in gastric cancer.

It is now believed that genes regulating apoptosis are also important variables in cancer development. Fas, a transmembrane protein of the tumour necrosis factor receptor family, is a key molecule for cell death signalling. The mutation of the primary structure of the Fas gene might also be one of the possible mechanisms that disrupt Fas-mediated apoptosis in tumour cells. The purpose of this study was to determine whether somatic mutation of the Fas gene could be involved in the tumourigenesis of gastric cancer. Polymerase chain reaction (PCR)-based loss of heterozygosity (LOH) analysis with two intragenic polymorphic markers, and mutation analysis for the entire coding regions of the Fas gene were performed in 43 cases of gastric cancer, using PCR-single-strand conformational polymorphism sequencing. Five (11.6%) missense mutations were detected, only in the death domain of the Fas gene. Although these mutations were observed only in intestinal-type gastric cancers, there was no statistically significant difference in the frequency of Fas mutation between intestinal- and diffuse-type gastric cancer (p=0.068). Nine LOH out of 22 informative cases were also detected with one or both markers (41%). Three of them demonstrated a somatic mutation in the remaining allele, indicating the inactivation of both alleles. These results suggest that genetic alterations of the Fas gene may not only be limited to gastric cancer cell protection through Fas resistance, but may also play an important role in tumour promotion and/or progression in a subset of gastric cancer.

Adenocarcinoma↗

Spatial temperature gradient capillary electrophoresis for DNA mutation detection.

A continuous spatial temperature gradient was established in capillary electrophoresis by using a simple temperature control device. The temperature profile along the capillary was predicted by theoretical calculations. A nearly linear spatial temperature gradient was established and applied to DNA mutation detection. By spanning a wide temperature range, it was possible to perform simultaneous heteroduplex analysis for various mutation types that have different melting temperatures.

DNA↗

Preliminary study of the analysis of oligogalacturonic acids by electrospray ionization mass spectrometry.

During systematic studies of the behavior of oligogalacturonic acids (OGAs) under different conditions using electrospray ionization mass spectrometry (ESI-MS), cation adduction, fragmentation and non-covalent binding were found to be the three major problems that compromised the analysis of OGAs by ESI-MS. By adjusting solution components, capillary temperature and capillary voltage in the ESI source, an optimized condition was found that gave a clean and clear spectrum of trigalacturonic acid. A direct injection ESI-MS technique based on the use of aqueous acetonitrile and acetic acid and triethylamine (TEA) as modifiers has been applied to analyze a mixture including mono-, di- and trigalacturonic acids, which will facilitate further applications of ESI-MS in the analysis of mixtures of OGAs.

Oligosaccharides↗

A cytotoxic secocycloartenoid from Abies koreana.

Two triterpenoids, 24-methylene-3,4-seco-cycloart-4(28)-en-3-oic acid (1) and 3-oxo-9beta-lanosta-7,22Z,24-trien-26,23-olide (6) were isolated from Abies koreana, together with beta-sitosterol (2), maltol (3), beta-sitosterol-O-beta-D-glucoside (4), and hexacosylferulate (5). The structures of the compounds were established based on the spectroscopic data. The cytotoxic activities of triterpenoids have been evaluated using the sulforhodamine B (SRB) method. Compound 1 showed moderate cytotoxicities against human lung carcinoma (A549), ovarian carcinoma (SK-OV-3), malignant melanoma (SK-MEL-2), and colon carcinoma (HCT-15) cell lines.

Abies↗

Islet cell autoimmunity and mitochondrial DNA mutation in Korean subjects with typical and atypical Type I diabetes.

AIMS/HYPOTHESIS: The 1997 American Diabetes Association classification of diabetes mellitus included a subset of Type I diabetic patients who do not need insulin for several years but eventually progress to complete insulin deficiency i. e. atypical Type I diabetes mellitus. In Caucasian populations, most Type I diabetic patients have auto-antibodies against islet cells. We examined the frequency of the auto-antibodies against islet cells and mitochondrial DNA 3243 mutation in Koreans with typical and atypical Type I diabetes mellitus. METHODS: We measured plasma C-peptide level in 1870 consecutive Korean diabetic patients. Of these, 56 patients had insulin deficiency (fasting and glucagon-stimulated plasma C-peptide concentrations < or = 0.2 nmol/l and < or = 0.32 nmol/l, respectively), and they were subdivided into typical (n = 26) and atypical Type I (insulin-dependent) diabetes mellitus (n = 30) according to clinical manifestation. Islet cell antibody was measured by indirect immunofluorescence. Anti-GAD antibody and anti-ICA512 antibody were measured by radioimmunoassay. Mitochondrial DNA 3243 mutation was detected using restriction enzyme Apa-I digestion of the amplified genomic DNA. RESULTS: The overall prevalence of auto-antibodies in the typical and atypical groups was 77% and 57%, respectively. Mitochondrial DNA 3243 mutation was found in 3 out of 30 (10%) of atypical Type I (insulin-dependent) diabetic patients but not in typical Type I (insulin-dependent) diabetic patients. CONCLUSION/INTERPRETATION: Autoimmunity might not be the only cause of progressive insulin deficiency in Koreans. Mitochondrial DNA mutation is another identifiable cause but the cause(s) of insulin deficiency in the remainder of Type I diabetic patients without autoimmunity is not clear.

Adolescent↗

A molecular genome scan analysis to identify chromosomal regions influencing economic traits in the pig. I. Growth and body composition.

Genome scans can be employed to identify chromosomal regions and eventually genes (quantitative trait loci or QTL) that control quantitative traits of economic importance. A three-generation resource family was developed by using two Berkshire grand sires and nine Yorkshire grand dams to detect QTL for growth and body composition traits in pigs. A total of 525 F2 progeny were produced from 65 matings. All F2 animals were phenotyped for birth weight, 16-day weight, growth rate, carcass weight, carcass length, back fat thickness, and loin eye area. Animals were genotyped for 125 microsatellite markers covering the genome. Least squares regression interval mapping was used for QTL detection. All carcass traits were adjusted for live weight at slaughter. A total of 16 significant QTL, as determined by a permutation test, were detected at the 5% chromosome-wise level for growth traits on Chromosomes (Chrs) 1, 2, 3, 4, 6, 7, 8, 9, 11, 13, 14, and X, of which two were significant at the 5% genome-wise level and two at the 1% genome-wise level (on Chrs 1, 2, and 4). For composition traits, 20 QTL were significant at the 5% chromosome-wise level (on Chrs 1, 4, 5, 6, 7, 12, 13, 14, 18), of which one was significant at the 5% genome-wise level and three were significant at the 1% genome-wise level (on Chrs 1, 5, and 7). For several QTL the favorable allele originated from the breed with the lower trait mean.

Animals↗

A molecular genome scan analysis to identify chromosomal regions influencing economic traits in the pig. II. Meat and muscle composition.

Molecular genetic markers can be used to identify chromosomal regions that contain quantitative trait loci (QTL) that control meat quality and muscle composition traits in farm animals. To study this in pigs, a resource family was generated from a cross between two Berkshire grand sires and nine Yorkshire grand dams. A total of 525 F2 progeny from 65 matings of F1 parents were produced. Phenotypic data on 28 meat quality traits (drip loss, water holding capacity, firmness, color, marbling, percentage cholesterol, ultimate pH, fiber type, and several sensory panel and cooking traits) were collected on the F2 animals. Animals were genotyped for 125 microsatellite markers covering the entire genome. Least squares regression interval mapping was used for QTL detection. Significance thresholds were determined by permutation tests. A total of 60 QTL were detected at the 5% chromosome level for meat quality traits, on Chrs 1, 2, 4, 5, 6, 7, 8, 10, 11, 12, 13, 14, 15, 17, 18, and X, of which 9 and 1 QTL were significant at the 5% and 1% genome-wise levels (on Chrs 1, 5, 12, 15, and 17), respectively.

Adipose Tissue↗

Correlation of plasma homocysteine and mitochondrial DNA content in peripheral blood in healthy women.

Hyperhomocysteinemia is an independent risk factor of cardiovascular disease and associated with insulin resistance, although their causal relationship remains unclear. A previous report has shown that high concentration of homocysteine damages mitochondrial gene expression, function and structure. As we found recently, the mitochondrial DNA (mtDNA) contents are inversely correlated with insulin resistance parameters. Thus there is possibility that plasma total homocysteine (tHcy) level is somewhat correlated with mtDNA content. Sixty healthy women (mean age 40.3+/-20.9 yr, range 18-78 yr) were recruited to investigate the correlation of plasma tHcy level and mtDNA content in peripheral blood. A significant negative correlation was found between plasma tHcy levels and mtDNA content (r=-0.507, P<0.01). Plasma tHcy and mtDNA content have an independent effect on each other and on insulin resistance (HOMA-insulin resistance (IR) score) respectively in multiple regression model. Plasma tHcy showed positive correlations with age (r=0.407), W/H ratio (r=0.370), total cholesterol (r=0.338), LDL-cholesterol (r=0.317) and insulin resistance (HOMA-IR score) (r=0.261); and a negative correlation with folate (r=-0.273). MtDNA content showed negative correlations with age (r=-0.407), BMI (r=-0.440), W/H ratio (r=-0.659), SBP (r=-0.350), total cholesterol (r=-0.340), triglyceride (r=-0.376), LDL-cholesterol (r=-0.349), fasting plasma insulin (r=-0.483), and insulin resistance (HOMA-IR score) (r=-0.423); and a positive correlation with folate (r=0.299). In this study, there was a significant inverse correlation between plasma tHcy level and mtDNA content. Further study will be warranted to elucidate the mechanism by which two factors are associated.

Adolescent↗

Peripheral blood mitochondrial DNA content is inversely correlated with insulin secretion during hyperglycemic clamp studies in healthy young men.

Abnormalities in mitochondrial DNA(mtDNA) have been implicated in the pathogenesis of diabetes mellitus. We recently reported that decreased mtDNA content precedes the development of diabetes mellitus and is associated with parameters of insulin resistance. In this study, we examined whether there is any relation between mtDNA content and insulin secretion. We compared the mtDNA content of peripheral blood leukocytes with the parameters of insulin secretion measured by hyperglycemic clamp in a group of healthy young men. There were statistically significant correlations between mtDNA content in peripheral blood and fasting plasma insulin (r=-0.43, P<0.05) and C-peptide levels (r=-0.44, P<0.05). MtDNA content also correlated negatively with acute insulin response(r=-0.48, P<0.05), late insulin response (r=-0.50, P<0.05) during hyperglycemic clamp and insulin secretion after glucagon stimulation (r=-0.60, P<0.01). mtDNA content in peripheral blood correlated negatively with homeostasis model (HOMA) insulin resistance (r=-0.45, P<0.05) although it did not correlate with the insulin insensitivity index (M/I) during hyperglycemic clamp. In summary, the mtDNA content of peripheral blood correlated negatively with indices of insulin resistance and insulin secretion in healthy young men. The compensatory response of pancreas beta cells to insulin resistance might contribute in part to increased insulin secretion in these subjects.

Adult↗

Method of proof and evidences for the concept that mitochondrial genome is a thrifty genome.

A hypothesis that mitochondrial genome itself is the thrifty genome has been advanced, as both its mutation, or quality, and amount in the peripheral blood, or quantity, are found to have close relationship to diabetes mellitus, or more specifically insulin resistance. Mere presence of relationship, however, does not establish cause-effect relation. Usually a genetic 'cause' is established, when a knockout of putative gene produces disease phenotype. However, absence of mitochondrial genome is incompatible with life. Here the argument advanced by Bradford Hill was applied to the proof of a hypothesis, as most of the data are from epidemiological observations; it is biologically plausible that both mtDNA quality and quantity are related to insulin resistance. The relationship had enough strength of association, dose-response relation, and consistency of relationship. Temporally association, decrease of mtDNA content preceding onset of diabetes, was also observed. Good hypothesis generates many predictions, which could be verified experimentally and which is biologically plausible.

Animals↗