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Biomedical subjects

H Han

Publications and source records attributed to H Han.

At least 37 records · Page 2Linked to original sources

Distribution of HLA-A, B alleles and polymorphisms of TAP and LMP genes in Korean patients with atopic dermatitis.

BACKGROUND: Atopic dermatitis has been seen to result from multifactorial inheritance, with interaction between genetic and environmental factors. The genetic association may differ according to the ethnic backgrounds. OBJECTIVE: The purpose of this study was to investigate the genetic factors in Korean atopic dermatitis patients by studying the human leucocyte antigen (HLA) class I association and polymorphisms of transporters associated with antigen presentation (TAP) and low-molecular-weight polypeptide (LMP) genes. METHODS: HLA-A and B genotyping was performed in 53 atopic dermatitis patients and 184 healthy controls using the standard microlymphocytotoxicity technique. TAP1, TAP2, LMP2, and LMP7 gene polymorphisms were anaylzed using the polymerase chain reaction (PCR)-single strand conformation polymorphism (SSCP), PCR-amplification refractory mutation system (ARMS), and PCR-restriction fragment length polymorphism (RFLP). RESULTS: Allele frequency of HLA-A24 was significantly increased in patients with atopic dermatitis compared to controls (P < 0.05). HLA-B alleles showed no differences in distribution between patients and controls. Genotype, phenotype, and allele frequencies of TAP1 gene also revealed no differences in distribution between patients and controls. Analysis of TAP2 gene polymorphisms showed increased frequencies of the TAP2*C allele and TAP2*A/TAP2*C genotype in atopic dermatitis patients compared to controls (P < 0.05). Distribution of LMP2 and LMP7 gene polymorphisms was similar for patients and controls. CONCLUSION: This study demonstrates an association of atopic dermatitis with HLA-A24 and TAP2*C alleles in Korean patients. Discrepancy with the previous reports might be related to different patient characteristics and ethnic variations.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

Serum cystatin C is an independent predictor of total homocysteine levels in stable Korean renal transplant recipients with normal serum creatinine.

OBJECTIVES: To examine the determinants of fasting plasma total homocysteine (tHcy) levels such as cystatin C, serum creatinine (SCr), estimated glomerular filtration rate (GFR) from Cockroft-Gault equation, albumin, plasma folate, vitamin B12, and pyridoxal-5'-phosphate (PLP) among Korean renal transplant recipients (RTR) with normal SCr levels (< or =1.4 mg/dL). DESIGN: Cross-sectional study. SETTING: Nephrology and Transplant Service, Catholic University Kangnam St. Mary's Hospital, Seoul, Korea. PARTICIPANTS: Fifty-one chronic stable Korean RTR with normal SCr levels (< or =1.4 mg/dL) 6 months or more following transplantation. MEASURES: Medical record review, anthropometric measurements, and overnight (10 to 14 hours) fasting blood samples for measurement of plasma tHcy, folate, vitamin B12, PLP, SCr, albumin, and cystatin C. RESULTS: General linear regression model including age, gender, vitamin status, and measurements of renal function showed that cystatin C and folate were independent predictors of tHcy levels. The partial regression coefficient for folate was -0.444 (P <.01) and for cystatin C, it was +0.334 (P <.05). SCr, estimated GFR, vitamin B12, PLP, age, and gender were not independent predictors of tHcy levels in this model. CONCLUSION: Both cystatin C and folate status were major independent determinants of fasting tHcy levels in the subgroup of Korean RTR with normal SCr.

Adult↗

A useful insight into 2 occlusal indexes: HLD(Md) and HLD(CalMod).

Maryland and California are among the states that have adopted the Handicapping Labiolingual Deviation (HLD) Index to determine eligibility for public funding of orthodontic treatment. As a result of several lawsuits against the state, California has developed and employed a modification of that index, called the HLD(CalMod) index. This study compared and contrasted the effects of analyzing prospective patients with the HLD(Md)-the HLD index as used in Maryland-and the HLD(CalMod) indexes. The specific aims of this study were to observe (1) the correlation between the 2 occlusal indexes in determining patients' eligibility for treatment with public funding, (2) the characteristics of patients eligible according to each index, and (3) the major contributing factors in assigning patients as eligible under each index. Initial study models of 313 patients from the orthodontic clinic at the University of Maryland were scored by both HLD(Md) and HLD(CalMod) indexes. According to the scores, the patients were divided into 2 groups-those with handicapping malocclusions and those without handicapping malocclusions. Scores from each index were analyzed by the Pearson product moment correlation. The Angle classification for each patient was also recorded with the HLD variables. The mean and the percentage of the total score for each variable were calculated. The following conclusions were obtained: (1) Of the 313 patients, 127 (41%) were approved for treatment by the HLD(Md) index, and 110 (35%) were approved for treatment by the HLD (CalMod) index.(2) The correlation between the 2 indexes (R = 0.78) was not very strong, indicating the power of the additional variables introduced in the HLD(CalMod) index. (3) The use of the HLD(CalMod) index directs more public funding toward patients with severe Class II malocclusions than does the HLD(Md) index.

Adolescent↗

Expression of multiple subtypes of muscarinic receptors and cellular distribution in the human heart.

Five isoforms of the muscarinic acetylcholine receptor (mAChR) have been identified by molecular cloning and designated m(1)-m(5), of which four correspond to the functional subtypes M(1), M(2), M(3), and M(4) in primary tissues. The presence of M(5) receptors in tissues remains uncertain. The present study was designed to explore the diversity and cellular distribution of various mAChR subtypes in human hearts. Competition binding of [N-methyl-(3)H]-scopolamine methyl chloride with various mAChR antagonists yielded data consistent with the presence of multiple subtypes (M(1)/M(2)/M(3)/M(5)) of mAChRs in both human atrial (HA) and ventricular (HV) tissues. Expression of mRNAs encoding all five subtypes was readily detected by reverse transcription-polymerase chain reaction in both HA and HV samples. Immunoblotting with subtype-specific antibodies confirmed the presence of M(1), M(2), M(3), and M(5), but not M(4), proteins in membrane preparations from both HA and HV. The protein levels of M(1) and M(2) were comparable between HA and HV. Although the density of M(3) appeared approximately 10-fold higher in HV than HA, that of M(5) was approximately 5 times lower in HV than in HA. Positive immunostaining of single ventricular myocytes by M(1), M(2), M(3), and M(5) antibodies, respectively, was consistently detected. Under confocal microscopy, M(5) showed characteristic localization to the intercalated discs, whereas other subtypes were more evenly distributed throughout the surface membrane. Our results provide the first molecular evidence for the presence of multiple subtypes of mAChR, including endogenous M(5) receptors, in human hearts and suggest that different subtypes have different tissue distributions and cellular localization.

Binding, Competitive↗

Regulation of phosphate uptake in primary cultured rabbit renal proximal tubule cells by glucocorticoids: evidence for nongenomic as well as genomic mechanisms.

We have investigated the nongenomic as well as the genomic effects of glucocorticoids on phosphate (Pi) uptake in primary rabbit renal proximal tubule cells (PTCs) and have defined the involved signaling pathways. In the present study, cortisol-BSA (cortisol-BSA) (>10(-9) M, 30 min) was found to inhibit Pi uptake in a time- and concentration-dependent manner. However, progesterone-BSA (P(4)-BSA), 17ss-estradiol-BSA (E(2)-BSA), testosterone-BSA (T(4)-BSA), aldosterone, P(4), E(2), and T(4) (10(-9) M, 1 h) had no effect on Pi uptake. In addition, cortisol-BSA (10(-9) M) did not affect either Na(+) uptake or alpha-methylglucopyranoside (alpha-MG) uptake. The cortisol-BSA-induced inhibition of Pi uptake was associated with a decrease in the V(max) for Pi uptake, rather than the K(m). The inhibitory effect of cortisol-BSA was not blocked either by actinomycin D (an inhibitor of transcription), cycloheximide (an inhibitor of translation), or classical glucocorticoid receptor antagonists (RU 486 or P(4)). The cortisol-BSA-induced inhibition of Pi uptake was blocked by two phospholipase C (PLC) inhibitors (neomycin or U73122), and two protein kinase C (PKC) inhibitors (staurosporine or bisindolylmaleimide I) but not by two adenylate cyclase/protein kinase A inhibitors [SQ 22536 (an adenylate cyclase inhibitor) or myristoylated protein kinase A inhibitor amide 14-22]. Furthermore, cortisol-BSA promoted the translocation of PKC from the cytosolic fraction to the membrane fraction, while having no effect on the activity of adenylate cyclase. Our observations may thus be interpreted as indicating that cortisol does indeed inhibit renal Pi uptake via a nongenomic mechanism, which involves the PLC/PKC pathway.

Animals↗

Posttraumatic stress disorder and extent of trauma exposure as correlates of medical problems and perceived health among women with childhood abuse.

This study examined the relative contributions of Posttraumatic Stress Disorder (PTSD) symptoms and the extent of trauma exposure as factors contributing to the current health status of childhood abuse survivors. Sixty-seven women with a history of familial childhood abuse (sexual and/or physical) and twenty-nine women with no abuse history were assessed on two distinct aspects of health status: reported number of medical problems and perceptions of overall health. Women with abuse were found to have a greater number of medical problems and poorer perceived physical well-being than the no abuse comparison group. Regression analyses of the women with abuse histories revealed that trauma exposure was a stronger predictor than PTSD symptoms of medical problems. PTSD symptoms, however, were better predictors of the experience of physical well-being than trauma exposure. These results suggest that the nature of a traumatic exposure, especially when there is repeated, cumulative trauma may be more significant to medical problems than the psychological symptoms of PTSD. Perceived health, however, appears to be predominantly influenced by psychological factors, suggesting the importance of these variables in the quality of life of multiply traumatized women.

Adolescent↗

[Establishment of suc2 signal sequence trap system].

The secretion of proteins depends on the signal peptide located to the N-terminal of the protein precursor. We established a genetic system in yeast to screen cDNA library for the signal peptide encoding sequences. To do it, we mutated genomic suc2 gene (encoding yeast invertase) of EGY48 by one-step gene disruption method, and got yeast cell lines without invertase expression (EGY48-delta suc). To get vector for library screening, we inserted suc2 gene encoding mature peptide of invertase downstream to yeast promoter P-ADH1, and multiple cloning sites for insertion of library is between suc2 and P-ADH1. EGY48-delta suc transformed with the vector can grow on the medium with glucose as carbon source, but not on the medium with raffinose. Signal peptide of suc2 and alpha chain of human interleukin-2 was fused in frame to suc2 gene, then the two resulting vectors were transformed into EGY48-delta suc, all the transformants can grow in the medium with either raffinose or glucose as carbon source. Hence, the system established here can discern cDNA encoding signal peptide from the one not encoding signal peptide.

Amino Acid Sequence↗

[Method on ozone generation with strong ionization discharge].

This paper presents the formed methods of strong ionization discharge of dielectric barrier and plasma chemical reaction process of ozone generation. Ozone combination and decomposition are controlled by electric field intensity and electron energy. Therefore, new technologies with thinner dielectric layers (230 microns) of model alpha Al2O3 and narrow discharge gap (110 microns) are introduced, and strong ionization discharge is gained which reduced field (E) and electron average energy are more than 400Td and 10 eV respectively. Ozone concentration reaches to 200 g/m3 and ozone producing efficiency is 100 g/(kW.h). Ozone generator of big yield and miniaturization with module assembled method is realized.

Electricity↗

[Cloning, sequence analysis of deacetoxycephalosporin C synthetase/hydroxylase gene cefEF].

Chromosomal DNA preparations were made form the fungi C. acremonium and A. chrysogenum respectively by a method using 1-Chloro methyl benzene solution. Then the cefEF gene of deacetoxycephalosporin C synthetase/hydroxylase was amplified by polymerase chain reaction. The cefEF gene could only be amplified from the chromosomal DNA of A. chrysogenum. This fragment about 1.0 kb was cloned into pGME-T vector and then sequenced. The sequence indicates that the cloned cefEF gene contains 999 nucleotides encoding for 333 amino acids and there are three bases and two amino acids different from those that have been reported. Both of the two amino acids are not in active site. The result also indicates that the reported C. acremonium or A. chrysogenum generating deacetoxycephalosporin C synthetase/hydroxylase corresponds with A. chrysogenum but not C. acremonium in China.

Base Sequence↗

High-performance polyethylene glycol-coated solid-phase microextraction fibers using sol-gel technology.

The sol-gel method is applied for the preparation of solid-phase microextraction (SPME) fibers. An electron microscopy experiment suggested a porous structure for Superox-4 (polyethylene glycol, PEG) coating. SPME-GC analyses provided evidence that the sol-gel fibers have some advantages, such as high velocities of mass transfer, efficient extraction rates. high thermal stability, long life span, and spacious range of application for both polar and non-polar analytes. Efficient SPME-GC analyses of benzene-toluene-ethylbenzene-xylenes, phenols, phthalic diesters, naphthalene congeners and pesticides were achieved using sol-gel-coated PEG fibers.

Chromatography, Gas↗

Expression of various multidrug resistance-associated protein (MRP) homologues in brain microvessel endothelial cells.

Multidrug resistance-associated protein (MRP) actively transports a broad range of anionic compounds out of the cell. To date, six different homologues of MRP (i.e. MRP1-MRP6) have been identified. The current study examines the expression of the various MRP homologues in both primary cultured bovine brain microvessel endothelial cells (BBMEC) and the capillary-enriched fraction from bovine brain homogenates. RT-PCR analysis demonstrated the presence of MRP1, MRP4, MRP5 and MRP6 in both BBMEC and the capillary-enriched fractions of brain homogenates. While low levels of MRP3 were detected in the BBMEC, it was not observed in the capillary-enriched fraction. In addition, RT-PCR and Western blot studies indicated an absence of MRP2 expression in both blood-brain barrier preparations. The presence of several different MRP homologues in the brain microvessel endothelial cells may be important in controlling the permeability of the blood-brain barrier to organic anions.

ATP-Binding Cassette Transporters↗

Inhibition of unwinding of G-quadruplex structures by Sgs1 helicase in the presence of N,N'-bis[2-(1-piperidino)ethyl]-3,4,9,10-perylenetetracarboxylic diimide, a G-quadruplex-interactive ligand.

N,N'-Bis[2-(1-piperidino)ethyl]-3,4,9,10-perylenetetracarboxylic diimide (PIPER), a perylene derivative, is a very potent and selective G-quadruplex DNA-interactive agent. It has been shown to inhibit DNA polymerase and telomerase by stacking externally to the G-tetrads in the G-quadruplex structures. Recently, we have demonstrated that this small molecule greatly accelerates the assembly of G-quadruplex structures in a cell-free system. In this report, we present data demonstrating that PIPER prevents the unwinding of G-quadruplex structures by yeast Sgs1 helicase. Sgs1 belongs to the RecQ DNA helicase family whose members include other G-quadruplex DNA unwinding helicases, such as human Bloom's syndrome and human Werner's syndrome helicases. PIPER specifically prevents the unwinding of G-quadruplex DNA but not duplex DNA by Sgs1. Competition experiments indicate that this inhibitory activity is due to the interaction of PIPER with G-quadruplex structures rather than the helicase itself. These results combined with previous studies suggest a possible mechanism of action for these G-quadruplex-interactive agents inside cells: they might induce G-quadruplex formation in G-rich regions on genomic DNA, stabilize these structures, and prevent them from being cleared by enzymes such as helicases. The G-quadruplex structures may, in turn, disrupt some critical cellular events such as DNA replication, transcription regulation, and telomere maintenance.

Anthracenes↗

Molecular phylogenetic study of the tribe Trypetini (Diptera: Tephritidae), using mitochondrial 16S ribosomal DNA sequences.

Monophyly and intratribal relationships of the tribe Trypetini were tested using mitochondrial 16S ribosomal RNA gene from 16 species of Trypetini, six tephritid outgroups, and two non-tephritid outgroups. The number of aligned sites was 1279 bp, but 1165 bp were used for analysis after excluding sites with missing data or gaps. Among these 1165 sites, 447 sites were variable and 305 were informative for parsimony analysis. Phylogenetic information was extracted from this data set using neighbor-joining and maximum parsimony methods and compared to a phylogenetic hypothesis proposed from the morphological literature. My molecular data suggest: (1) monophyly of the tribe Trypetini; (2) monophyly of the Chetostoma group; (3) monophyly of the Trypeta group; and (4) non-monophyletic nature of the previous concept of Hemilea. I suggest that more sensitive genetic markers with less A+T bias are needed to bring about finer resolution within the tribe Trypetini, and additional tephritid lineages should be sampled to identify the sister-group relationship of the tribe Trypetini.

Journal Article↗

Cloning and characterization of the 5'-flanking region for the mouse phospholipase C-delta1 gene.

To date, little is known about the molecular mechanisms controlling the regulation of phospholipase C-delta1 (PLC-delta1) gene expression. To understand the mechanisms responsible for the regulation of PLC-delta1 gene expression, the 5'-flanking region of the mouse PLC-delta1 gene was isolated from a mouse genomic DNA library. Primer extension analysis revealed that there is a single transcriptional start site located at 127 bases upstream from the translation start codon in the mouse PLC-delta1 gene. DNA sequence analysis showed that the sequence around the transcriptional start site is very GC-rich and has no TATA or CAAT boxes. Transient expression of a luciferase reporter gene under the control of serially deleted 5'-flanking sequences revealed that the 160-base-pair region from -622 to -462 upstream of the transcriptional start site includes a positive cis-acting element(s) for the efficient expression of the PLC-delta1 gene. Gel retardation analysis suggests that multiple transcription factors bind to separate sites on the promoter region. Based on these results, our study suggests that the minimal essential region located at -622 to +70 is fully sufficient to confer high-level transcriptional activity and contains high-affinity binding elements for multiple transcription factors.

Animals↗

Overexpression of rapsyn modifies the intracellular trafficking of acetylcholine receptors.

Rapsyn is a protein that interacts with the cytoplasmic face of the nicotinic acetylcholine receptors (AChR) to cluster them within postsynaptic membrane of muscle. Here we show that intracellular AChRs are also affected by rapsyn. When rapsyn was co-transfected with AChR into QT-6 fibroblasts, (125)I-alpha-bungarotoxin binding indicated a reduction in the fraction of AChRs expressed on the cell surface, compared to cells expressing AChRs alone. Double fluorescent labeling showed that intracellular AChRs accumulated in patches at the cell periphery, beneath rapsyn-associated cell surface AChR clusters. These patches were observed even when cells were grown in medium containing excess unlabelled alpha-bungarotoxin to mask internalized AChRs, suggesting that they arose from hindered trafficking of newly formed AChRs to the cell surface. Similarly, in the muscle cell line, C2, overexpression of rapsyn resulted in the co-localization of aggregates of intracellular alpha-bungarotoxin binding sites with rapsyn beneath cell surface AChR microaggregates. The results indicate that rapsyn can modify the trafficking of AChRs within the cell and suggest a role in selectively targeting newly synthesized intracellular AChRs to the postsynaptic membrane.

Animals↗

Human cytomegalovirus gene products US3 and US6 down-regulate trophoblast class I MHC molecules.

The epidemiological correlation between human CMV (HCMV) infection and spontaneous fetal loss has been suggested, but the underlying mechanism is not well understood. Fetal cytotrophoblasts, which are in direct contact with the maternal immune system in the uterus during pregnancy, do not express HLA-A and HLA-B, but express the nonclassical class I HLA-G and HLA-C. It has been shown that both HLA-G and HLA-C are capable of inhibiting NK-mediated cell lysis. In our present study, using human trophoblast cell lines as well as other cell lines stably transfected with the human class I genes, we have demonstrated that HCMV US3 and US6 down-regulate the cell-surface expression of both HLA-G and HLA-C by two different mechanisms. HCMV US3 physically associates with both trophoblast class I MHC species, retaining them in the endoplasmic reticulum. In contrast, HCMV US6 inhibits peptide transport by TAP and thus specifically the intracellular trafficking of class I molecules. Therefore, these findings suggest for the first time a possible molecular mechanism underlying HCMV-related spontaneous pregnancy loss.

3T3 Cells↗

Association of HLA with Vogt-Koyanagi-Harada syndrome in Koreans.

PURPOSE: To study the distribution of human leukocyte antigen HLA-A/B antigens and HLA-DR/-DQ/-DP alleles and to investigate the immunogenetic background of Korean patients with Vogt-Koyanagi-Harada (VKH) syndrome and clinical course with different types of HLA. METHODS: Human leukocyte antigen typings were performed in 18 Korean patients with VKH syndrome and in 128 healthy control subjects. HLA-A/B loci serologic typing was performed according to the standard microlymphocytotoxicity technique. DNA was extracted through the salting out method, and HLA-DR phenotyping and HLA-DR4, HLA-DQ, and HLA-DP subtyping were performed with the polymerase chain reaction-sequence specific oligonucleotide probe (PCR-SSOP) method. RESULTS: Among HLA-A/B antigens typed by the standard microlymphocytotoxicity method, the frequencies of HLA-A31 (RR = 6.1, P<1x10(-2)) and HLA-B55 (RR = 15.8, P<.05) were significantly increased in the patient group compared with the control group. Among HLA-DR/-DQ/-DP alleles subtyped by DNA methods, the frequencies of HLA-DRB1*04 (RR = 45.1, P<1x10(-7)) and HLA-DRB1*07 (RR = 3.2, P<.05) were significantly increased. However, significant decreases in HLA-DRB1*08 (RR = .1, P<.05), HLA-DRB1*13 (RR = .1, P<.05), and HLA-DRB1*14 (RR = .1, P<.05) frequencies were observed. The result of HLA-DR, HLA-DQ, and HLA-DP subtyping showed the significant increase in DRB1*0405 (RR = 45.1, P<1x10(-7)), DQA1*0302 (RR = 12.0, P<1x10(-4)), DQB1*0303 (RR = 5.0, P<1x10(-2)), DQB1*0401 (RR = 18.9, P<1x 10-6), and DPB1*0501 (RR = 3.8, P<.05). However, significant decreases in DQA1*0101 (RR = .1, P< .05), DQA10102 (RR = .1, P<1x10(-2)), DQA1*0103 (RR = .1, P<.05), DQA1*0501 (RR = .1, P<1x10(-2)), DQB1*0301 (RR = .1, P<.05), DQB1*0601 (RR = .1, P<.05), DPB1*0201 (RR = .3, P<.05), and DPB1*0401 (RR = .1, P<.05) frequencies were also observed. In patients with DRB1*0405 itself or HLA-DRB1*0405-DQA1*0302-DQB1*0401 haplotype, a reduction in visual acuity and ocular complications was common. CONCLUSIONS: These results suggest that HLA-DRB1*0405 itself or HLA-DRB1*0405-DQA1*0302-DQB1*0401 haplotype is greatly increased and may play the most important role in the development and the clinical course of VKH syndrome in Korean patients.

Alleles↗

G-quadruplexes as targets for drug design.

G-quadruplexes are a family of secondary DNA structures formed in the presence of monovalent cations that consist of four-stranded structures in which Hoogsteen base-pairing stabilizes G-tetrad structures. These structures are proposed to exist in vivo, although direct confirmatory evidence is lacking. Guanine-rich regions of DNA capable of forming G-quadruplex structures are found in a variety of chromosomal regions, including telomeres and promoter regions of DNA. In this review, we describe the design of three separate groups of G-quadruplex-interactive compounds and their interaction with G-quadruplex DNA. Using the first group of compounds (anthraquinones), we describe experiments that provide the proof of concept that a G-quadruplex is required for inhibition of telomerase. Using the second group of compounds (perylenes), we describe the structure of a G-quadruplex-ligand complex and its effect on the dynamics of formation and enzymatic unwinding of the quadruplex. For the third group of compounds (porphyrins), we describe the experiments that relate the biological effects to their interactions with G-quadruplexes.

Base Sequence↗