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Biomedical subjects

K S Park

Publications and source records attributed to K S Park.

At least 19 recordsLinked to original sources

Chemokine regulation of human megakaryocytopoiesis.

We have previously shown that platelet factor 4 (PF4), a platelet-specific CXC chemokine, can directly and specifically inhibit human megakaryocyte colony formation. We therefore hypothesized that PF4 might function as a negative autocrine regulator of megakaryocytopoiesis. Herein we present additional studies characterizing the inhibitory effect of CXC chemokines on human megakaryocyte development. We first corroborated our initial studies by showing that recombinant human (rH) PF4, like the native protein, inhibited megakaryocytopoiesis. We then examined the inhibitory properties of other CXC family members. Neutrophil activating peptide-2 (NAP-2), a naturally occurring N-terminally cleaved beta TG peptide, was found to inhibit megakaryocytopoiesis with two to three orders of magnitude greater potency than PF4. Structure function studies showed that an N-terminal mutation, which eliminated NAP-2's neutrophil activating properties (NAP-2E2-->A), also abrogated its ability to inhibit megakaryocyte development. Further investigations of this type demonstrated that a chimeric PF4 protein (AELR/PF4) in which PF4's N-terminus was replaced with the first four amino acids of NAP-2 was also a potent inhibitor of megakaryocytopoiesis. Interleukin (IL)-8, another CXC chemokine, and three CC chemokines (macrophage inhibitory protein-1 alpha [MIP-1 alpha], MIP-1 beta, and C10) also specifically inhibited megakaryocyte colony formation at NAP-2 equivalent doses. CXC and CC chemokine inhibition was additive suggesting that the effects might be mediated through a common pathway. The inhibitory effects of NAP-2 and MIP-1 alpha could not be overcome by adding physiologically relevant amounts of recombinant human megakaryocyte growth and development factor (MGDR) (50 ng/mL) to the cultures. Using Northern blot and reverse transcriptase-polymerase chain reaction (RT-PCR) based analyses, we documented mRNA expression of IL-8 receptor isoforms alpha and beta in total platelet RNA and in normal human megakaryocytes, respectively. Based on these results, we hypothesize that chemokines play a physiologic role in regulating megakaryocytopoiesis. Because chemokines are elaborated by ancillary marrow cells, both autocrine and paracrine growth control is suggested, the effects of which might be exerted, in part, through alpha and beta IL-8 receptors.

Amino Acid Sequence

Improvement of thermal stability of subtilisin J by changing the primary autolysis site.

The thermostability of subtilisin J, an extracellular serine protease secreted from Bacillus stearothermophilus, has been improved by changing the primary autolysis site of the Asp-49 mutant protein. Previously we have shown that the Asp-49 mutant protein has proteolytic activity, but so unstable that it was primarily autolyzed in Tyr-58-Gln-59 peptide bond during cultivation (Jang et al. Biochim. Biophys. Acta. 1162, 233-235 1993). In the present study, to mitigate the autolytic degradation and increase the thermostability, we deleted the Tyr-58 residue using the Asp-49 mutant as a template. This mutant (Asp-49/delta Tyr-58 mutant) protein showed an improved resistance to heat treatment without changing the catalytic efficiency of the enzyme. These results show that change of primary autolysis site can stabilize the subtilisin.

Amino Acid Sequence

Serum acetone and liver acetone monooxygenase activity in pregnant rats, fetuses, and neonates: reversible pretranslational reduction of cytochrome P450IIE1 (P450IIE1) during pregnancy.

Serum acetone in neonates was found to increase from < 20 microM at 20 days gestation to 377 +/- 107 microM 1 day after birth. This increase in acetone occurs concurrently with the initial expression of liver P450IIE1 in rat (Song et al., 1986, J. Biol. Chem. 261:16689-16697). Treating pregnant rats with drinking water containing 1% acetone did not result in a premature induction of liver acetone monooxygenase, a P450IIE1 catalyzed activity, in fetuses sacrificed at 20 days gestation. The data indicate that increased serum acetone levels are not responsible for the initial induction of P450IIE1 in neonates. However, acetone monooxygenase activity in their mothers was significantly less than acetone-treated females which were not pregnant, indicating a reduced activity of P450IIE1 during pregnancy although its induction by acetone was still observed. Acetone monooxygenase activity in 20 day pregnant rats given untreated drinking water was also less than in nongravid rats. These findings are supported by immunoblot data which show significant and progressive reductions in liver P450IIE1 in both untreated and acetone-treated rats during pregnancy. Northern mRNA blot analysis further revealed that the decreases in P450IIE1 activity and protein content were mainly due to pretranslational suppression with reduced level of its mRNA. However, their levels rapidly returned to the control level after parturition (within 1 day). Repeated administration of several exogenous hormones (estriol, pregnanediol, thyroxine) or peptide hormones (placental lactogen, prolactin, chorionic gonadotropin) failed to suppress P450IIE1 in nonpregnant rats, indicating a possibility of another factor(s) responsible for the P450IIE1 suppression during pregnancy.

Acetone

A survey of trace elements in fresh-water fish and rice along the Han River by neutron activation analysis.

For a case study of environmental pollution, radiochemical neutron activation analysis (RNAA) was applied to the crucian and rice collected along the Han River. The crucian was analyzed for three times in 1973, 1987, and 1990. Sixteen trace elements (Hg, Cd, As, Br, Cu, Na, K, Se, Cr, Hf, Rb, Fe, Zn, Co, La, and Cs) were determined by RNAA using distillation and diethyldithiocarbamate extraction methods. Contents of Na, K, Se, Hf, Fe, Zn, and Co were almost constant regardless of the sampling place and year. The contents of the other elements showed increasing trends down river, especially in the first investigation. At the lower part of the river, the contents showed decreasing trends with the time of sampling, especially during the first two investigations. These trends were typical for Hg and Cd. Rice was analyzed by the same method for 12 elements, and the results showed no regional trends, but have decreased after 1973.

Animals

The effect of obesity on fibrinolytic activity and plasma lipoprotein (a) levels in patients with type 2 diabetes mellitus in Korea.

To determine whether previously reported abnormalities in fibrinolytic activity and plasma lipoprotein (a) levels could reflect obesity rather than diabetes per se, plasma concentrations of tissue-type plasminogen activator (t-PA), type 1 plasminogen activator inhibitor (PAI-1), and lipoprotein (a) (Lp (a)) were investigated in sixty-four type 2 diabetic patients (56.1 +/- 9.5 years; body mass index, 24.6 +/- 3.3 kg/m2) and thirty-two control subjects (57.9 +/- 8.9 years; body mass index, 24.6 +/- 3.4 kg/m2). Both the plasma t-PA and PAI-1 antigen levels were similar between the diabetic group (10.6 +/- 3.8 ng/ml; 27.7 +/- 11.6 ng/ml) and the control group (12.2 +/- 3.5 ng/ml; 27.7 +/- 9.6 ng/ml). The PAI-1 levels were evenly distributed from 5.93 to 52.7 ng/ml in diabetic patients. The difference of Lp (a) levels between the two groups was negligible (the diabetic group, median 11 mg/dl (range 0-72 mg/dl); the control group, median 13 mg/dl (range 0-55 mg/dl)). Significant correlations between PAI-1 levels and body mass index (BMI) were observed in both groups. In the diabetic group, PAI-1 levels also correlated with fasting C-peptide levels (r = 0.54, P < 0.01) and serum triglyceride levels (r = 0.28, P < 0.05). However, we could not find a significant association between either t-PA or PAI-1 levels and Lp (a) levels in the diabetic and control groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult

Identification of Borrelia burgdorferi isolated in Korea using outer surface protein A (OspA) serotyping system.

Two characteristic strains (935T, 934U) of B. burgdorferi isolated from Ixodes persulcatus and a wild rodent (Apodemus agrarius) in Korea were selected and analyzed by an immunoblot method using the monoclonal antibodies directed to different epitopes of outer surface protein A (OspA). The reactive pattern of strain 934U with these monoclonal antibodies was identical to that of strains belonging to B. afzelii and that of strain 935T was different from other isolates. Monoclonal antibody (5TEE3) which is specific to strain 935T did not react with any other Western and Japanese isolates. So, it was suggested that there exist at least two groups of B. burgdorferi in Korea. One could be classified as B. afzelii and the other is a divergent group from three known species of B. burgdorferi sensu stricto, B. garinii and B. afzelii.

Animals

Exophytic adenocarcinoma of the stomach: CT findings.

OBJECTIVE: Exophytic adenocarcinomas of the stomach are tumors that have large extraluminal portions. Exophytic growth of gastric carcinoma is rare, and such tumor may be confused with gastric leiomyosarcoma. The purpose of this study was to assess the CT findings of exophytic adenocarcinoma of the stomach and to determine their value in distinguishing between it and gastric leiomyosarcoma. MATERIALS AND METHODS: Twenty patients with exophytic adenocarcinomas of the stomach confirmed by surgery or endoscopic biopsy were included in the study. CT scans were assessed retrospectively for the size, location, and character of the mass and the presence of adjacent thickening of the gastric wall and outlet obstruction. RESULTS: The masses were from 5 to 14 cm in diameter (mean, 9 cm). The mass was in the gastric antrum in 11 cases, the body of the stomach in six cases, the body and antrum in two cases, and the gastric fundus and body in one case. CT showed an exogastric mass with a variable degree of internal low density or necrosis in all 20 cases. Thickening of the gastric wall adjacent to the mass appeared to be a specific finding of exogastric adenocarcinoma (n = 14); it was not seen in reported cases of leiomyosarcoma. Gastric outlet obstruction was seen in four cases, and ulcer was detected in eight cases. The degree of low density or necrosis within the mass was minimal in eight cases, moderate in seven cases, and severe in five cases. CONCLUSION: Our results suggest that CT findings of thickening of the gastric wall adjacent to an exogastric mass, representing spread of cancer to adjacent gastric wall, and/or gastric outlet obstruction are typical of exophytic adenocarcinoma of the stomach and allow distinction between it and gastric leiomyosarcoma.

Adenocarcinoma

Mechanism of decrease in heart rate by peripheral dopaminergic D2-receptors.

We performed this study in order to verify the heart rate decrease caused by the D2-receptor on cardiac sympathetic nerve endings and its relation to the concentration of norepinephrine in synaptic clefts. Sprague-Dawley rats were pithed and the heart rate was increased either by electrical stimulation of the cardiac accelerator nerve or by intravenous infusion of norepinephrine, tyramine, or isoproterenol. Increased heart rate by electrical stimulation of cardiac accelerator nerve was dose-dependently lowered by lisuride and its effect was blocked by pretreatment with sulpiride but not with yohimbine and SCH 23390. Also, the heart rate was decreased in a dose-dependent manner by clonidine and this effect was blocked by pretreatment with yohimbine, but not with sulpiride. For increased heart rate by infusion of norepinephrine, tyramine, or isoproterenol, the heart rate lowering effect of lisuride was more marked in the norepinephrine-and tyramine-infusion groups, in which the intrasynaptic concentration of norepinephrine was elevated, compared to the isoproterenol-infusion group, in which intrasynaptic concentration of norepinephrine was not elevated. In conclusion, there is a D2-receptor on the cardiac sympathetic nerve endings which decreases the heart rate and is different from the presynaptic alpha 2-receptor. Also, the heart rate lowering effect via stimulation of the D2-receptor by lisuride was more marked with increased concentration of norepinephrine in the synaptic cleft.

Animals

Association of complement C4 and HLA-DR alleles with systemic lupus erythematosus in Koreans.

OBJECTIVE: To examine the association of complement C4 and HLA-DR to systemic lupus erythematosus (SLE) susceptibility in Korea. METHODS: Complement C4 protein typing was carried out by immunofixation and immunoblotting methods using EDTA-plasma from 60 patients with SLE and 72 healthy controls. Restriction fragment length polymorphism analysis of C4 genes was also carried out using TaqI or HindIII for restriction enzymes. HLA-DR was determined by polymerase chain reaction amplification with sequence specific primers using genomic DNA from 67 patients with SLE and 72 healthy controls. RESULTS: The frequency of the C4AQ0 allele was significantly higher in the patients with SLE than in controls (41.7 vs 25.0%, p < 0.05). The deletion of the C4A gene commonly found in Caucasian patients with SLE was not observed in any patients. For HLA-DR, a significant increase of the haplotype DRB1*1501 was observed in the patients (26.9 vs 12.5%, p < 0.05) and DR9 was also significantly increased (23.9 vs 11.1%, p < 0.05). An increase in each DR2 and DR9 was independent of an increase in C4AQ0. The frequencies of DR2 and DR9 were significantly decreased in patients with renal involvement and alopecia, respectively. CONCLUSION: Our data suggested that the presence of C4AQ0 allele, DRB1*1501-DRB5*0101 haplotype and DR9 contributed to susceptibility to SLE in Koreans and that Korean SLE is based on a different genetic background from Caucasian patients.

Adult

The asparaginyl-tRNA synthetase gene encodes one of the complementing factors for thermosensitive translation in the Escherichia coli mutant strain, N4316.

Escherichia coli strain N4316 is a mutant that exhibits temperature-sensitive growth at 43 degrees C and temperature-sensitive translation in vivo and in vitro. Extracts of the mutant produce an aberrant pattern of translation products of MS2 bacteriophage RNA. Previous work has shown that a protein, called 'rescue', isolated from the parental strain partly corrects the defective translation in vitro. Here we report the purification to homogeneity of a second factor from ribosomal eluates of the wild-type parental strain; the purified protein is a homodimer of 54 kDa. The partial sequence of the second protein was determined, and a recombinant plasmid was isolated based on its ability to complement the temperature-sensitive growth phenotype of the mutant at the non-permissive temperatures. The cloned gene was sequenced, mapped to the 20.9-min region of the E. coli chromosome and shown to code for a 466-amino-acid protein with a molecular mass of 52 kDa. Analysis of the DNA sequence and the correspondence to that of the partial protein sequence has identified the complementing factor as asparaginyl-tRNA synthetase. Marker rescue experiments indicate that the asnS mutation in N4316 resides within the motif 2 domain of the synthetase. A potential role of this synthetase in restoring normal protein synthesis with respect to ribosomal frameshifting, read-through of nonsense codons and protein copy number is discussed.

Amino Acid Sequence

Pre-translational induction of pentoxyresorufin O-depentylase by pyridine.

Pentoxyresorufin O-depentylase activity, mainly associated with phenobarbital-inducible cytochrome P450IIB1 (designated CYP2B1), was increased after a single treatment of pyridine (250 mg/kg, i.p.), and further increased by repeated treatments for 5 days. The catalytic activity and immunoreactive protein of CYP2B recognized by polyclonal antibodies were significantly induced by a relatively high dose of pyridine (250 mg/kg, i.p.) while ethanol-inducible cytochrome P450IIE1 (CYP2E1) could be induced by a low dosage (25 mg/kg, i.p.). Unlike CYP2E1 induction without changing its mRNA level, the induction of CYP2B by pyridine was accompanied by an elevation of its mRNA, indicating a pre-translational activation of this enzyme. These results indicate that pyridine induces various isozymes of cytochromes P450 by different induction mechanisms.

Aniline Hydroxylase

Allotypes of the fourth component of complement in Koreans.

The analysis of genetic polymorphism in C4 was performed on EDTA-plasma from 169 healthy unrelated Koreans. Plasma samples were subjected to high-voltage agarose gel electrophoresis followed by immunofixation. C4B allotypes were further detected by a hemolytic overlay method. The allele frequencies of C4A and C4B were as follows; for C4A, C4A*3 = 0.6099, C4A*4 = 0.1702, C4A*Q0 = 0.1525, C4A*2 = 0.0461, and C4A*R = 0.0213; for C4B, C4B*1 = 0.6406, C4B*2 = 0.2740, C4B*5 = 0.0569, C4B*Q0 = 0.0178, and C4B*R = 0.0107. C4A*3 and C4B*1 were among the most common alleles at each locus. C4A*6 was not detected in this study, but this allele is relatively common in both Caucasoid and Negroid populations. C4B*5 is a common allele in Asian, which is rare in Caucasoids and Negroids. C4B*5 appeared to be a characteristic allele of Oriental. In the C4A locus, five individuals with duplicated allotypes (three C4A 3,3 + 2, one C4A 4,3 + 2, and one C4A 3,3 + 3) were observed, and in the C4B locus, one individual with duplicated allotype (C4B 1,1 + 1) was detected.

Alleles

Relationships between %Hb F or %G gamma and the haplotypes in the beta-globin gene cluster in the normal adult Japanese and Korean populations.

Haplotypes or subhaplotypes in the beta-globin gene cluster were examined in 31 normal Japanese and 21 Korean families. Major haplotypes were VII, V, and I, which shared a common subhaplotype [+----], and occurred in 70% of the Japanese and 68% of the Koreans. Three haploypes which shared a common subhaplotype [-+-++], bearing the XmnI site 5' to the G gamma-globin gene, occurred in 13% of the Japanese and 7.3% of the Koreans. One subhaplotype [-++-+], which is closely linked to the A gamma T-globin gene, had an incidence of 6.6% in the Japanese and 20% in the Koreans. Rare subhaplotypes [---++], and [- +] which has been observed mainly in Melanesians and Polynesians, and a very rare subhaplotype [-----] were observed. The present study suggests that high or low G gamma-globin chain production is closely related to subhaplotypes [-+-++] and [+----], respectively, among the normal adult population.

Adult

The XmnI site 5' to the G gamma-globin gene polymorphism and its relationship to %Hb F and %G gamma in normal Japanese and Korean adults.

The ratio of fetal hemoglobin to total hemoglobin (%Hb F), the ratio of G gamma to total gamma globin (%G gamma), and the polymorphism of the XmnI site at -158 base pairs from the cap site of the G gamma-globin gene were examined in normal unrelated Japanese (n = 113) and Korean (n = 44) adults. The frequency of the presence of the XmnI site was 0.15 in the Japanese and 0.16 in the Korean population. There were statistically significant differences in the %G gamma values of the Japanese between those +/+ and those +/- or -/- at the XmnI site (p less than 0.01). The Korean %G gamma values showed a statistically significant difference (p less than 0.01) between those +/- and those -/-. The presence of the XmnI site was significantly associated with the elevation of G gamma-globin chain synthesis, but this relationship was not necessarily absolute. The absence of the XmnI site in an adult with a gamma-globin gene triplication (G gamma AG gamma A gamma/G gamma A gamma) more or less reduced the level of G gamma-globin chain synthesis, but the presence of the XmnI site in an adult with a gamma-globin gene deletion (GA gamma/G gamma A gamma) had no effects on the proportion of the two gamma-globin chains.

Adult

Brazilin protects cultured rat hepatocytes from BrCCl3-induced toxicity.

Brazilin, the main constituent of Caesalpinia sappan, is an antioxidative substance that has catechol moiety in its chemical structure. Considering the antioxidant-activity of brazilin, it was expected to have protective effects on the toxicities of radical generating chemicals. The incubation of rat hepatocytes with BrCCl3 resulted in significant increase in lipid peroxidation, leakage of cytoplasmic enzymes and cytoplasmic glutathione depletion. The BrCCl3-induced toxicities on hepatocytes were reduced by the treatment of brazilin. Brazilin has been also proved to have a protective effect on the BrCCl3-induced depression of microsomal calcium sequestration activity. These results indicate that brazilin plays a protective role in BrCCl3-induced hepatocyte injury of the rat.

Alanine Transaminase

Post-translational reduction of cytochrome P450IIE by CCl4, its substrate.

The molecular mechanism of cytochrome P450IIE reduction by CCl4 was reexamined by measuring its enzyme activity, immunoreactive protein contents, and mRNA levels. Aniline hydroxylase and the amounts of immunoreactive P450IIE were rapidly decreased in a time-dependent manner after a single dose of CCl4. No changes were observed in the amounts of immunoreactive P450IIC and P450IA despite significant decreases decrease in their catalytic activities. However, the decreases in P450IIE enzyme activity and immunoreactive protein by CCl4 were not accompanied by a decline in its mRNA level. The data thus suggested a post-translational reduction of P450IIE by CCl4, probably due to specific destruction of the P450IIE protein by its own substrate rather than heme moiety.

Aniline Hydroxylase

Intra-abdominal fat is associated with decreased insulin sensitivity in healthy young men.

To distinguish the relative role of intra-abdominal and subcutaneous abdominal fat in metabolic aberrations in upper body fat localization, we measured the relationship between regional fat distribution and insulin sensitivity in nine young men (28.6 +/- 0.7 years; body mass index [BMI], 24.7 +/- 1.3 kg/m2). Regional fat distribution was measured by anthropometric measurement and computed tomography (CT). Insulin sensitivity was measured by euglycemic hyperinsulinemic glucose clamp. Insulin sensitivity, expressed as the ratio of rate of glucose utilization to the mean plasma insulin concentration during the second hour of glucose clamp (M/I) was negatively correlated with BMI (r = -.91, P less than .001), waist to hip girth ratio (WHR) (r = -.80, P less than .01), subcutaneous abdominal fat area (r = -.90, P less than .001), and intra-abdominal fat area (r = -.88, P less than .01). Stepwise forward regression analysis showed that in addition to BMI, intra-abdominal fat area was a significant correlate of decrease in insulin sensitivity. These findings suggest that intra-abdominal fat play an important role in decreasing insulin sensitivity, even in healthy young men.

Abdomen