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Biomedical subjects

H Park

Publications and source records attributed to H Park.

At least 181 records · Page 10Linked to original sources
PubMed1999

Th2 responses to inactivated influenza virus can Be converted to Th1 responses and facilitate recovery from heterosubtypic virus infection.

Immunization with live influenza virus expands Th1 memory cells and facilitates more rapid recovery after heterosubtypic virus challenge. Immunization with inactivated virus generates a Th2 response and does not lead to heterosubtypic immunity. Creation of a Th1 priming environment by the inclusion of interleukin (IL)-12 with antibodies to IL-4 converted the response against inactivated virus to a Th1 response that was able to facilitate virus clearance upon heterosubtypic virus challenge. Evaluation of memory responses of mice immunized by the various protocols demonstrated that the type of immunization imprints T cell memory, dictating the nature of the response to subsequent infection. After live virus challenge, expansion of Th1 cells seems to facilitate the generation of cytotoxic T lymphocytes from naïve precursors. This latter finding may be the mechanism by which inactivated virus immunization in a Th1 cytokine context mediates heterosubtypic immunity.

Animals↗
PubMed1999

Expression and characterization of bioactive human thrombopoietin in the milk of transgenic mice.

Human thrombopoietin (hTPO) is the primary physiological regulator of platelet production and plays a pivotal role in promoting the proliferation and maturation of megakaryocytic progenitor cells and megakaryocytes. In this study, transgenic mice were produced harboring either full-length or the erythropoietin (EPO)-like amino-terminal domain of hTPO cDNA sequences fused to the regulatory elements of the bovine beta-casein gene. The transgene RNA was expressed exclusively in the mammary glands of eight transgenic mice, and a trace amount of the transgene was also found in the lungs of one mouse. The full-length form induced efficient expression of the protein with the highest expression level of 1500 microg/ml; however, the EPO-like domain alone expressed the protein at <0.1 microg/ml. The proteins from the two recombinant cDNAs have apparent molecular weights of about 74 and 17 kDa, due to glycosylation in the case of the full-length cDNA. Cell proliferation assay in vitro indicated that both of the recombinant forms stimulated proliferation of the TPO-dependent BaF3-Mpl cells. A positive correlation appeared between the amount of TPO in the milk of lactating animals and their blood platelet levels. About a twofold increase in platelet numbers in the blood was observed after direct subcutaneous injection of the recombinant hTPO at the level of 30 microg/kg of body weight. On the basis of these results, we anticipate that the recombinant hTPO produced efficiently in milk of transgenic mice will have the same activities as the native hTPO in a few in vivo as well as in vitro biochemical aspects.

Animals↗
PubMed1999

Dynamic behavior of driven interfaces in models with two absorbing states.

We study the dynamics of an interface (active domain) between different absorbing regions in models with two absorbing states in one dimension: probabilistic cellular automata models and interacting monomer-dimer models. These models exhibit a continuous transition from an active phase into an absorbing phase, which belongs to the directed Ising (DI) universality class. In the active phase, the interface spreads ballistically into the absorbing regions and the interface width diverges linearly in time. Approaching the critical point, the spreading velocity of the interface vanishes algebraically with a DI critical exponent. Introducing a symmetry-breaking field h that prefers one absorbing state over the other drives the interface to move asymmetrically toward the unpreferred absorbing region. In Monte Carlo simulations, we find that the spreading velocity of this driven interface shows a discontinuous jump at criticality. We explain that this unusual behavior is due to a finite relaxation time in the absorbing phase. The crossover behavior from the symmetric case (DI class) to the asymmetric case (directed percolation class) is also studied. We find the scaling dimension of the symmetry-breaking field y(h)=1.21(5).

Journal Article↗
PubMed1999

Active width at a slanted active boundary in directed percolation.

The width W of the active region around an active moving wall in a directed percolation process diverges at the percolation threshold p(c) as W approximately Aepsilon(-nu( parallel)) ln(epsilon(0)/epsilon), with epsilon=p(c)-p, epsilon(0) a constant, and nu( parallel)=1.734 the critical exponent of the characteristic time needed to reach the stationary state xi( parallel) approximately epsilon(-nu(parallel)). The logarithmic factor arises from screening the statistically independent needle shaped subclusters in the active region. Numerical data confirm this scaling behavior.

Journal Article↗
PubMed1999

Velocity and wall shear stress patterns in the human right coronary artery.

Blood flow dynamics in the human right coronary artery have not been adequately quantified despite the clinical significance of coronary atherosclerosis. In this study, a technique was developed to construct a rigid flow model from a cast of a human right coronary artery. A laser photochromic method was used to characterize the velocity and wall shear stress patterns. The flow conditions include steady flow at Reynolds numbers of 500 and 1000 as well as unsteady flow with Womersley parameter and peak Reynolds number of 1.82 and 750, respectively. Characterization of the three-dimensional geometry of the artery revealed that the largest spatial variation in curvature occurred within the almost branch-free proximal region, with the greatest curvature existing along the acute margin of the heart. In the proximal segment, high shear stresses were observed on the outer wall and lower, but not negative, stresses along the inner wall. Low shear stress on the inner wall may be related to the preferential localization of atherosclerosis in the proximal segment of the right coronary artery. However, it is possible that the large difference between the outer and inner wall shear stresses may also be involved.

Blood Flow Velocity↗
PubMed1999

The hypervariable domain of the murine leukemia virus surface protein tolerates large insertions and deletions, enabling development of a retroviral particle display system.

The surface proteins (SU) of murine type-C retroviruses have a central hypervariable domain devoid of cysteine and rich in proline. This 41-amino-acid region of Friend ecotropic murine leukemia virus SU was shown to be highly tolerant of insertions and deletions. Viruses in which either the N-terminal 30 amino acids or the C-terminal 22 amino acids of this region were replaced by the 7-amino-acid sequence ASAVAGA were fully infectious. Insertions of this 7-amino-acid sequence at the N terminus, center, and the C terminus of the hypervariable domain had little effect on envelope protein (Env) function, while this insertion at a position 10 amino acids following the N terminus partially destabilized the association between the SU and transmembrane subunits of Env. Large, complex domains (either a 252-amino-acid single-chain antibody binding domain [scFv] or a 96-amino-acid V1/V2 domain of HIV-1 SU containing eight N-linked glycosylation sites and two disulfides) did not interfere with Env function when inserted in the center or C-terminal portions of the hypervariable domain. The scFv domain inserted into the C-terminal region of the hypervariable domain was shown to mediate binding of antigen to viral particles, demonstrating that it folded into the active conformation and was displayed on the surface of the virion. Both positive and negative enrichment of virions expressing the V1/V2 sequence were achieved by using a monoclonal antibody specific for a conformational epitope presented by the inserted sequence. These results indicated that the hypervariable domain of Friend ecotropic SU does not contain any specific sequence or structure that is essential for Env function and demonstrated that insertions into this domain can be used to extend particle display methodologies to complex protein domains that require expression in eukaryotic cells for glycosylation and proper folding.

Amino Acid Sequence↗
PubMed1999

Evidence for peroxynitrite as a signaling molecule in flow-dependent activation of c-Jun NH(2)-terminal kinase.

The c-Jun NH(2)-terminal kinase (JNK), also known as stress-activated protein kinase, is a mitogen-activated protein kinase that determines cell survival in response to environmental stress. Activation of JNK involves redox-sensitive mechanisms and physiological stimuli such as shear stress, the dragging force generated by blood flow over the endothelium. Laminar shear stress has antiatherogenic properties and controls structure and function of endothelial cells by mechanisms including production of nitric oxide (NO) and superoxide (O(-)(2)). Here we show that both NO and O(-)(2) are required for activation of JNK by shear stress in endothelial cells. The present study also demonstrates that exposure of endothelial cells to shear stress increases tyrosine nitration, a marker of reactive nitrogen species formation. Furthermore, inhibitors or scavengers of NO, O(-)(2), or reactive nitrogen species prevented shear-dependent increase in tyrosine nitration and activation of JNK. Peroxynitrite alone, added to cells as a bolus or generated over 60 min by 3-morpholinosydnonimine, also activates JNK. These results suggest that reactive nitrogen species, in this case most likely peroxynitrite, act as signaling molecules in the mechanoactivation of JNK.

Animals↗
PubMed1999

No relationship between serum lipoprotein(a) and albumin concentrations in patients with acute phase response.

The aim of this study was to investigate the relationship between lipoprotein(a) [Lp(a)] and albumin concentrations in the serum of patients with acute phase response (APR). We have compared the Lp(a) concentrations and apolipoprotein [apo(a)] phenotypes of 40 controls with those of 40 APR patients with normoalbuminaemia and 40 APR patients with hypoalbuminaemia. We have also compared concentrations of haptoglobin (Hp) and alpha 1-antitrypsin (AAT) containing a high sialic acid content, similar to Lp(a). The mean serum Lp(a) concentration (SD) of the 40 controls was 0.190 (0.142) g/L. The mean serum Lp(a) concentration was 0.358 (0.257) g/L (P < 0.001) in 80 APR patients. However, there was no difference in serum Lp(a) concentrations between the APR patients with hypoalbuminaemia [0.353 (0.268) g/L] and the APR patients with normoalbuminaemia [0.362 (0.249) g/L]. No significant difference was found in the distributions of apo(a) phenotypes between the controls, the APR patients with hypoalbuminaemia, and the APR patients with normoalbuminaemia (P = 0.183). In the APR patients, the serum concentrations of AAT and Hp were respectively 2.709 (0.822) g/L and 2.631 (1.340) g/L, whereas those of normal controls were respectively 1.422 (0.219) g/L (P < 0.001) and 0.956 (0.442) g/L (P < 0.001). In conclusion, the Lp(a) is one of the acute phase reactants whose synthesis concurrently increases with other APRs, especially those with a high sialic acid content. The increase of the serum Lp(a) concentrations in the APR patients is not related to serum albumin concentration.

Acute-Phase Reaction↗
PubMed1999

The risk groups for coronary heart disease in Koreans. Assessment by distribution of serum lipid concentrations.

Coronary heart disease is the most severe form of disease caused by atherosclerosis; and there is a strong relationship between serum lipid concentrations and atherosclerosis. By decade of life, means, standard deviations and selected percentiles were calculated according to sex for serum concentrations of total cholesterol, low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C) and triglyceride in 69,563 Koreans. The prevalence of dyslipoproteinemias which were related to high risk of coronary heart disease were assessed. The mean concentrations of serum total cholesterol and LDL-C in the Korean population were 5.02 mmol/l and 3.00 mmol/l. The 75th and 90th percentile concentrations of total cholesterol were 5.59 mmol/l and 6.24 mmol/l. The mean concentrations of serum triglyceride and HDL-C were 1.58 mmol/l and 1.30 mmol/l. The hyperlipoproteinemia type IV (4.8%) was the most frequent, followed by Type IIa (4.6%), hypoalphalipoproteinemia (3.3%), and type IIb (0.2%). According to the National Cholesterol Education Program Adult Treatment Panel II, 4.1% of Korean adults needed the initial drug therapy and 10.8% the initial dietary therapy for hypercholesterolemia. The age and sex-specific treatment guidelines for hypercholesterolemia would make it possible that early intervention could be applied to atherosclerosis in Korean adults.

Adolescent↗
PubMed1999

Level of in utero cocaine exposure and neonatal ultrasound findings.

OBJECTIVE: To assess whether there is an association between the level of in utero cocaine exposure and findings on neonatal cranial ultrasound, controlling for potentially confounding variables. STUDY DESIGN: In a prospective longitudinal study, three cocaine exposure groups were defined by maternal report and infant meconium assay: unexposed, heavier cocaine exposure (>75th percentile self-reported days of use or of meconium benzoylecogonine concentration) or lighter cocaine exposure (all others). Neonatal ultrasounds from 241 well, term infants were read by a single radiologist who was masked to the exposure group. RESULTS: Infants with lighter cocaine exposure did not differ from the unexposed infants on any ultrasound findings. After controlling for infant gender, gestational age, and birth weight z scores and for maternal parity, blood pressure in labor, ethnicity, and use of cigarettes, alcohol, and marijuana during pregnancy, the more heavily cocaine-exposed infants were more likely than the unexposed infants to show subependymal hemorrhage in the caudothalamic groove (covariate adjusted odds ratio: 3.88; 95% confidence interval: 1.45, 10.35). CONCLUSIONS: This is the first study to demonstrate that ultrasound findings suggestive of vascular injury to the neonatal central nervous system are related to the level of prenatal cocaine exposure. Inconsistency in previous research in identifying an association between prenatal cocaine exposure and neonatal cranial ultrasound findings may reflect failure to consider dose effects.

Cocaine↗
PubMed1998

Plasma membrane cholesterol is a key molecule in shear stress-dependent activation of extracellular signal-regulated kinase.

Shear stress, the dragging force generated by fluid flow, differentially activates extracellular signal-regulated kinase (ERK) and c-Jun NH2-terminal kinase (JNK) in bovine aortic endothelial cells (BAEC) (Jo, H., Sipos, K., Go, Y. M., Law, R., Rong, J., and McDonald, J. M. (1997) J. Biol. Chem. 272, 1395-1401). Here, we examine whether cholesterol-enriched compartments in the plasma membrane are responsible for such differential regulation. Pretreatment of BAEC with a cholesterol-binding antibiotic, filipin, did not inhibit shear-dependent activation of JNK. In contrast, filipin and other membrane-permeable cholesterol-binding agents (digitonin and nystatin), but not the lipid-binding agent xylazine, inhibited shear-dependent activation of ERK. The effect of cholesterol-binding drugs did not appear to be due to membrane permeabilization, since treatment of BAEC with a detergent, Triton X-100 which also permeabilizes membranes, did not inhibit shear-dependent activation of ERK. Furthermore, shear-dependent activation of ERK, but not JNK, was inhibited by cyclodextrin, a membrane-impermeable cholesterol-binding agent, which removes cell-surface cholesterol. Moreover, the effects of cyclodextrin were prevented by adding cholesterol during the incubation. These results indicate that cholesterol or cholesterol-sensitive compartments in the plasma membrane play a selective and essential role in activation of ERK, but not JNK, by shear stress. Although exposure to shear stress (1 h) increased the number of caveolae by 3-fold, treatment with filipin had no effect in either control or shear-exposed cells suggesting that caveolae density per se is not a crucial determinant in shear-dependent ERK activation. In summary, the current study suggests that cholesterol-sensitive microdomains in the plasma membrane, such as caveolae-like domains, play a critical role in differential activation of ERK and JNK by shear stress.

Animals↗
PubMed1998

NMR structure of the histidine kinase domain of the E. coli osmosensor EnvZ.

Bacteria live in capricious environments, in which they must continuously sense external conditions in order to adjust their shape, motility and physiology. The histidine-aspartate phosphorelay signal-transduction system (also known as the two-component system) is important in cellular adaptation to environmental changes in both prokaryotes and lower eukaryotes. In this system, protein histidine kinases function as sensors and signal transducers. The Escherichia coli osmosensor, EnvZ, is a transmembrane protein with histidine kinase activity in its cytoplasmic region. The cytoplasmic region contains two functional domains: domain A (residues 223-289) contains the conserved histidine residue (H243), a site of autophosphorylation as well as transphosphorylation to the conserved D55 residue of response regulator OmpR, whereas domain B (residues 290-450) encloses several highly conserved regions (G1, G2, F and N boxes) and is able to phosphorylate H243. Here we present the solution structure of domain B, the catalytic core of EnvZ. This core has a novel protein kinase structure, distinct from the serine/threonine/tyrosine kinase fold, with unanticipated similarities to both heatshock protein 90 and DNA gyrase B.

Amino Acid Sequence↗
PubMed1998

Peptides glycosylated in the endoplasmic reticulum of yeast are subsequently deglycosylated by a soluble peptide: N-glycanase activity.

Several lines of evidence suggest that soluble peptide:N-glycanase (PNGase) is involved in the quality control system for newly synthesized glycoproteins in mammalian cells. Here we report the occurrence of a soluble PNGase activity in Saccharomyces cerevisiae. The enzyme, which was recovered in the cytosolic fraction, has a neutral pH optimum, and dithiothreitol is required for activity. All of these properties were similar to those of earlier described for mammalian PNGases. Interestingly, the yeast enzyme activity was found to be present almost exclusively in cells in stationary phase; little activity was detected in logarithmic growth phase cells. Upon incubation of a glycosylatable peptide R-Asn-X-Thr-R' with permeabilized yeast spheroplasts, we detected formation of both glycosylated peptide and the peptide product expected from PNGase-mediated deglycosylation of this glycopeptide, namely, R-Asp-X-Thr-R'. Recent findings that yeast have an active system for the retrograde transport of unfolded (glyco)proteins and glycopeptides out of the endoplasmic reticulum (ER) into the cytosol raise the possibility that this PNGase may participate in an early step in degradation of these molecules following their export from the ER.

Amidohydrolases↗
PubMed1998

Two-domain reconstitution of a functional protein histidine kinase.

In prokaryotes, in the absence of protein serine/threonine/tyrosine kinases, protein histidine kinases play a major role in signal transduction involved in cellular adaptation to various environmental changes and stresses. Histidine kinases phosphorylate their cognate response regulators at a specific aspartic acid residue with ATP in response to particular environmental signals. In this His-Asp phosphorelay signal transduction system, it is still unknown how the histidine kinase exerts its enzymatic function. Here we demonstrate that the cytoplasmic kinase domain of EnvZ, a transmembrane osmosensor of Escherichia coli can be further divided into two distinct functional subdomains: subdomain A [EnvZ(C). (223-289); 67 residues] and subdomain B [EnvZ(C).(290-450); 161 residues]. Subdomain A, with a high helical content, contains the autophosphorylation site, H-243, and forms a stable dimer having the recognition site for OmpR, the cognate response regulator of EnvZ. Subdomain B, an alpha/beta-protein, exists as a monomer. When mixed, the two subdomains reconstitute the kinase function to phosphorylate subdomain A at His-243 in the presence of ATP. Subsequently, the phosphorylated subdomain A is able to transfer its phosphate group to OmpR. The two-domain structure of this histidine kinase provides an insight into the structural arrangement of the enzyme and its transphosphorylation mechanism.

Bacterial Outer Membrane Proteins↗
PubMed1998

A glycosidase antibody elicited against a chair-like transition state analog by in vitro immunization.

Antibodies were generated against the positively charged chair-like glycosidase inhibitor nojirimycin by in vitro immunization. A number of catalytic antibodies were isolated, one of which catalyzes the hydrolysis of p-nitrophenyl beta-D-glucopyranoside 3 with a rate enhancement (kcat/kuncat) of 10(5) M over the HOAC-catalyzed reaction. The antibody discriminates modifications in the pyranoside ring of substrate 3 at the C2, C4, and the anomeric positions. The pH dependence of the reaction and chemical modification studies suggest the presence of an active-site Asp or Glu residue that may function as a general acid. This study further defines those requirements necessary to generate antibodies that efficiently cleave glycosidic bonds.

1-Deoxynojirimycin↗
PubMed1998

Linkage disequilibrium of the Apo AI-CIII-AIV gene cluster and their relationship to plasma triglyceride, apolipoprotein AI and CIII levels in Koreans.

More than 20 different RFLPs have been described in the apo AI-CIII-AVI gene cluster and their association with several dyslipidemias has been claimed in different populations. So far, however, the results of their association have been largely inconclusive. We analyzed six previously described RFLPs (Paul-Hayase et al., 1992) in the apo AI-CII-AIV gene cluster by using PCR-amplified DNAs in 132 healthy subjects to document the relationship between the RFLPs and dyslipoproteinemias. The polymorphic sites were located in the promoter region of the apo AI gene (G to A substitution, MspI), in the 3' flanking region of the apo AI gene (PstI), in the 3' noncoding region of the apo CIII gene (SstI), in the first intron of the apo CIII gene (PvuIIa), in the intergenic region of the apo CIII and apo AIV genes (PvuIIb), and in the second intron of the apo AIV gene (XbaI), respectively. The allele frequencies of PstI polymorphism showed no ethnic difference. However, in G to A substitution sites (MspI), Koreans had a lower G allele frequency than that of Caucasians. Among Oriental ethnic groups there were no significant differences in the allele frequencies of SstI and PvuIIa RFLPs, whereas significant differences existed between individuals of Oriental, Caucasian, and/or African origins. Koreans were monomorphic for PvuIIb and XbaI restriction sites. Pairwise linkage disequilibrium analysis revealed two pairs of significant negative linkage disequilibriums (MspI-SstI and SstI-PvuIIa). Although there were no statistical significances, the S2 allele of SstI polymorphism and the G allele of MspI polymorphism seemed to be associated with hypertriglyceridemia. Thus linkage of the G allele of MspI polymorphism and the S2 allele of SstI polymorphism could give an additive effect on an increment of triglyceride levels. We did not find any association between the SstI polymorphism and apo CIII levels. In conclusion, there were no statistically significant relationships detected between the apo AI-CIII-AVI gene cluster polymorphisms and the levels of various lipid parameters.

Adolescent↗
PubMed1998

Identification of functionally important residues of human thrombopoietin.

Thrombopoietin (TPO) is a megakaryocyte growth and differentiation factor. It consists of a characteristic two domain structure. The amino-terminal domain of TPO has a sequence homology with erythropoietin and is required for the binding and activation of its receptor c-Mpl. To determine the functionally important regions interacting with its receptor, a series of site-directed mutants of TPO were constructed based on a three-dimensional model of the amino-terminal domain. Two strategies of mutagenesis were employed: 1) nonnative N-linked glycosylation scan of 12 residues predicted to be on the surface, and 2) alanine replacement scan of mostly charged 44 amino acid residues. Each TPO mutein was transiently expressed in COS7 cells, and the specific bioactivity of the TPO protein secreted into the culture medium was measured using a recombinant BaF3 cell line expressing human c-Mpl. Four alanine substitutions at Arg10, Pro42, Glu50, and Lys138 nearly or completely abolished the activity, whereas the mutation at Arg14 slightly decreased the activity, suggesting that these residues are functionally important in interacting with its receptor. These residues mapped to helix A, loop AB, and helix D. Sequence comparison between human TPO and other mammalian TPO showed that the identified residues are completely conserved among the species. However, unlike the recent report on the mutational analysis of TPO, alanine substitutions at Lys52, Lys59, Arg136, and Arg140 did not affect the TPO activity significantly in our system. The identified receptor binding regions of TPO are analogous to those of human growth hormone and erythropoietin. Based on the similarity of these three cytokines, we propose that Lys138 of helix D and Pro42 and Glu50 of loop AB may constitute one binding region, whereas Arg10 and Lys14 of helix A may constitute the other binding region to dimerize the receptors.

Alanine↗