Biomedical subjects

B C Park

Publications and source records attributed to B C Park.

At least 19 recordsLinked to original sources

PubMed2006

Isolation and characterization of outer root sheath melanocytes of human hair follicles.

BACKGROUND: Outer root sheath melanocytes (ORSM) are not yet routinely cultured and their biology is not known in detail because of their relatively low numbers in the hair follicle and their limited proliferative capacity in in vitro culture in routine media. OBJECTIVES: To develop a method for culturing ORSM more easily and to investigate the length of telomeres and antigenic characteristics of ORSM compared with epidermal melanocytes (EM). METHODS: Hair follicles were obtained from three Korean individuals during hair transplantation surgery. Single-cell suspensions of the outer root sheath were made and cultured in melanocyte growth medium with stem cell factor. After 21 days, second-passage outer root sheath keratinocytes (ORSK) (2 x 10(4) mL(-1) MGM) were added into the culture plates. We studied the proliferation pattern, morphological and antigenic characteristics of ORSM for each passage of cultured cells, and observed ORSM telomere length. RESULTS: We established an ORSM culture method using ORSK. Two morphologically different ORSM types were obtained in the primary cultures. At the end of primary culture, ORSM appeared as whitish-cream pellets. The proliferation pattern of ORSM showed a sigmoidal shape, the accumulated numbers of population doublings showed a plateau after approximately 5 months, and senescence occurred at approximately 33 +/- 5 accumulated population doublings. The length of ORSM telomeres continued to shorten as the cells proliferated. In contrast, EM showed a marked proliferation from the early proliferation period which formed a plateau pattern towards the later period, and the number of accumulated population doublings was estimated to be 18 +/- 5 after 2 months. ORSM in the primary culture reacted variably with l-dihydroxyphenylalanine (DOPA): some cells were DOPA negative, some DOPA positive. There were some different antigenic expressions of microphthalmia-associated transcription factor (MITF) showing cytoplasmic expression in ORSM and nuclear expression in EM. By nuclear extraction and Western blotting, we showed that MITF expression of ORSM was marked in the cytoplasm and minimal in the nucleus. Antigenic expression of MITF and Bcl-2 gradually decreased with increasing passage number, whereas tyrosinase-related protein-1 expression did not change. CONCLUSIONS: Culture of ORSM requires ORSK or ORSK-related factors; ORSM have greater proliferation potential and show different MITF antigenic expression compared with EM; and the length of ORSM telomeres shortens with repeated proliferation.

Adult↗

PubMed2002

High bifurcation of median nerve at the wrist causing common digital nerve injury in endoscopic carpal tunnel release.

A 45-year-old right-handed woman was treated by endoscopic carpal tunnel release under local anaesthetic. When cutting the proximal half of the flexor retinaculum with the retrograde blade, she complained of an electric shock-like pain in her middle and ring fingers. Open exploration of the carpal tunnel was performed and a laceration of the middle/ring common digital nerve, which bifurcated at the unusually high level of the wrist crease, was found.

Carpal Tunnel Syndrome↗

PubMed2001

Primary unreamed intramedullary nailing for open fractures of the tibia.

Forty-six open tibial fractures (42 patients) were treated by primary unreamed intramedullary nailing, with debridement of open wounds and treatment of soft tissue. According to Gustilo-Anderson classification there were 18 grade I cases, 18 grade II cases and 10 grade III cases. The incidence of infection was low with two cases of superficial infection and one of deep infection. The mean time for union was 21.9 weeks and the rate of nonunion was 10.8%. There was no significant difference in the mean time to union, infection rate and rate of nonunion with different site and grade of fracture, but there was a longer union time and a higher rate of nonunion in complex and comminuted fractures. Unreamed intramedullary nailing, with appropriate soft tissue treatment, gives good results in the treatment of open tibial fractures including grade III.

Adolescent↗

PubMed2001

Non-combustible waste vitrification with plasma torch melter.

Non-combustible radioactive wastes generated from Nuclear Power Plants (NPPs) are composed of concrete, glass, asbestos, metal, sand, soil, spent filters, etc. The melting tests for concrete, glass, sand, and spent filters were carried out using a 60 kW plasma torch system. The surrogate wastes were prepared for the tests. Non-radioactive Co and Cs were added to the surrogates in order to simulate the radioactive waste. Several kinds of surrogate prepared by their own mixture or by single waste were melted with the plasma torch system to produce glassy waste forms. The characteristics of glassy waste forms were examined for the volume reduction factor (VRF) and the leach rate. The VRFs were estimated through the density measurement of the surrogates and the glassy waste forms, and were turned out to be 1.2-2.4. The EPA (Environmental Protection Agency) Toxicity Characteristic Leaching Procedure (TCLP) was used to determine the leach resistance for As, Ba, Hg, Pb, Cd, Cr, Se, Co, and Cs. The leaching index was calculated using the total content of each element in both the waste forms and the leachant. The TCLP tests resulted in that the leach rates for all elements except Co and Cs were lower than those of the Universal Treatment Standard (UTS) limits. There were no UTS limits for Co and Cs, and their leach rate & index from the experiments were resulted in around 10 times higher than those of other elements.

Chemical Phenomena↗

PubMed2001

Mitogenic and metabolic effects of type I IGF receptor overexpression in insulin receptor-deficient hepatocytes.

We have previously shown that hepatocytes lacking insulin receptors (Ir-/-) fail to mediate metabolic responses, such as stimulation of glycogen synthesis, while retaining the ability to proliferate in response to IGFs. In this study we have asked whether overexpression of type I IGF receptors would rescue the metabolic response of Ir-/- hepatocytes. After IGF-I stimulation, insulin receptor substrate-1 and -2 phosphorylation and PI3K activity were restored to levels similar to or greater than those seen in wild-type cells. Rates of cell proliferation in response to IGF-I increased approximately 2-fold, whereas glycogen synthesis was restored to wild-type levels, but was comparatively smaller than that elicited by overexpression of insulin receptors. In summary, overexpression of IGF-I receptors in Ir-/- hepatocytes normalized insulin receptor substrate-2 phosphorylation and glycogen synthesis to wild-type levels, whereas it increased cell proliferation above wild-type levels. Moreover, stimulation of glycogen synthesis was submaximal compared with the effect of insulin receptor overexpression. We conclude that IGF-I receptors are more efficiently coupled to cell proliferation than insulin receptors, but are less potent than insulin receptors in stimulating glycogen synthesis. The data are consistent with the possibility that there exist intrinsic signaling differences between insulin and IGF-I receptors.

Animals↗

PubMed2001

Genetics of type 2 diabetes: insight from targeted mouse mutants.

Diabetes affects millions of people worldwide, and its chronic complications are a leading cause of death in many industrialized countries. In a minority of patients, diabetes is brought about by the auto-immune destruction of insulin-producing pancreatic beta cells (Type 1 diabetes). In the vast majority of patients, diabetes is brought about by a combination of genetic and environmental factors that affect the organism's ability to respond to insulin (Type 2 diabetes). This impairment is due to a complex abnormality involving insulin action at the periphery and insulin production in the beta cell. Genetic factors play a key role in the development of type 2 diabetes. However, the inheritance of diabetes is non-Mendelian in nature, due to genetic heterogeneity, polygenic pathogenesis and incomplete penetrance. For these reasons, many laboratories have developed "designer" mice bearing targeted mutations in genes of the insulin action and insulin secretion pathways in order to develop a better model for the inheritance and pathogenesis of type 2 diabetes. These mutant mice are beginning to challenge established paradigms in the pathogenesis of type 2 diabetes and to shed light onto the genetic interactions underlying its complex inheritance. Here we review recent progress in the field and assess its impact on human studies of the genetics, prevention and treatment of type 2 diabetes.

Animals↗

PubMed2000

Isoleucine imbalance using selected mixtures of imbalancing amino acids in diets of the broiler chick.

Three experiments were conducted to determine the isoleucine requirement of a broiler from the time of hatch to 16 d of age. Chicks in the experiments were fed an isoleucine-limiting diet composed of wheat and peanut meal as the primary protein sources; this diet was used to investigate various mixtures of amino acids as imbalancing agents for isoleucine. The isoleucine requirement for maximum weight gain and feed efficiency was determined on the basis of broken-line regression analysis to be 0.63 to 0.65% of the diet or 3.28 to 3.38% of dietary protein. A similar diet, marginally limiting in isoleucine, was used to investigate the response of chicks to the addition of various mixtures of amino acids to the diet. Chicks that received a 5% dietary addition of 11 amino acids consisting of equimolar concentrations of leucine, valine, histidine, methionine, phenylalanine, tryptophan, tyrosine, alanine, glycine, serine, and threonine had significantly lower weight gain and feed consumption and a higher feed conversion ratio than did chicks fed the basal diet. These adverse effects were only partly prevented by an isoleucine supplement. The large neutral amino acids, histidine, methionine, phenylalanine, tryptophan, and tyrosine, accounted for most of the effect of the mixture. No effect of a mixture of leucine and valine or a mixture of the small neutral amino acids (alanine, glycine, serine, and threonine) at the same concentrations as those in the mixture of the 11 amino acids was observed. Lysine and arginine were the only two indispensable amino acids not present in the mixture of 11 amino acids. A subsequent experiment demonstrated that these amino acids did not become co-limiting with isoleucine when the diet was imbalanced with the amino acid mixture. These results indicate that an isoleucine imbalance in chicks is readily precipitated by excessive dietary concentrations of large neutral amino acids in diets that are otherwise marginally adequate in isoleucine.

Aging↗

PubMed1999

Egr-1 mediates transcriptional activation of IGF-II gene in response to hypoxia.

We have previously reported that the exposure of human HepG2 cells to hypoxic conditions results in the overexpression of human insulin-like growth factor II (IGF-II) mRNA whose size is 6.0 kb. This particular size of IGF-II mRNA is transcribed under the control of the IGF-II P3 promoter. In the present study, to delineate the molecular mechanism for the activation of the IGF-II gene, we examined the induction of P3 promoter activity in HepG2 cells by hypoxia in the transient expression system. In this system, hypoxia induced a linear increase within 24 h in the expression of luciferase that was driven by the IGF-II P3 promoter. To further delineate which factors mediate this response, the expression pattern of regulators of the P3 promoter, Egr-1, Sp1, and WT1, were analyzed by reverse transcription-PCR and Northern blot analysis. We found that hypoxia increased the expression of Egr-1 but not of Sp1. In contrast, the level of WT1, a repressor of IGF-II expression, was markedly decreased during hypoxia. The mRNA stability assay revealed that the induction of transcription is the mechanism of underlying Egr-1 mRNA elevation. We then investigated the effects of hypoxia on the DNA binding activity of Egr-1. Both electrophoretic mobility shift assay and supershift assay demonstrated that the DNA binding activity of the Egr-1 protein was increased by hypoxia. In addition, the level of Egr-1 protein was also increased under the hypoxia as determined by Western blot analysis. Cotransfection of HepG2 cells with an Egr-1 expression vector and an IGF-II P3 promoter-luciferase reporter plasmid showed that the transcription of IGF-II was activated by Egr-1 in a dose-dependent manner. Moreover, the elevation of IGF-II P3 promoter activity was induced synergistically by the cotreatment of hypoxia with Egr-1 overexpression. Deletion of sequences in the IGF-II P3 promoter containing Egr-1 binding sites did not respond to hypoxic stress. Taken together, these data strongly indicate that hypoxia-induced IGF-II expression in HepG2 cells is due to the enhanced activity of Egr-1 on the IGF-II P3 promoter and that the Egr-1 binding site in the IGF-II P3 promoter is essential for the transcriptional regulation of IGF-II under hypoxic conditions.

Carcinoma, Hepatocellular↗

PubMed1999

Identification by chimera formation and site-selected mutagenesis of a key amino acid residue involved in determining stereospecificity of guinea pig 3-hydroxysteroid sulfotransferase isoforms.

The 3-hydroxysteroid sulfotransferases that have been isolated and cloned from humans and rodents appear to have broad substrate specificities. In the guinea pig, however, two 3-hydroxysteroid sulfotransferases have been isolated that function according to an innate stereospecificity: the alpha-isoform acts on steroids with a 3-hydroxyl group oriented in the alpha position, whereas the beta-isoform acts on steroids where the 3-hydroxyl group is in a beta orientation. To examine the structural basis for this remarkable stereoselectivity, chimeras of the two enzymes, which are 87% identical, were constructed. A chimera consisting of the NH(2)-terminal 91 amino acids of the alpha-isoform and the COOH-terminal 196 amino acids of the beta-isoform displayed activity similar to that of the alpha-isoform. Site-selected mutagenesis of this 3alpha/beta-hydroxysteroid sulfotransferase chimera involving the 12 amino acid differences that exist between the two isoforms within the 91 amino acid NH(2)-terminal region revealed that the amino acid residue at position 51 plays a fundamental role in determining the stereospecificity exhibited by the alpha- and beta-isoforms, i.e. if residue 51 is an asparagine, alpha activity predominates, whereas if an isoleucine is in that position, beta activity prevails.

Amino Acid Sequence↗

PubMed1999

Differential signaling of insulin and IGF-1 receptors to glycogen synthesis in murine hepatocytes.

We have used SV40-transformed hepatocytes from insulin receptor-deficient mice (-/-) and normal mice (WT) to investigate the different abilities of insulin and IGF-1 receptors to stimulate glycogen synthesis. We report that insulin receptors are more potent than IGF-1 receptors in stimulating glycogen synthesis. Both receptors stimulate glycogen synthesis in a PI 3-kinase-dependent manner, but only the effect of insulin receptors is partially rapamycin-dependent. Insulin and IGF-1 receptors activate Akt to a similar extent, whereas GSK-3 inactivation in response to IGF-1 is considerably lower in both -/- and WT cells, compared to the effect of insulin in WT cells. The findings indicate that (i) the potency of insulin and IGF-1 receptors in stimulating glycogen synthesis correlates with their ability to inactivate GSK-3, (ii) the extent of GSK-3 inactivation does not correlate with the extent of Akt activation mediated by insulin or IGF-1 receptors, indicating that the effect of insulin on GSK-3 requires additional kinases, and (iii) the pathways required for insulin stimulation of glycogen synthesis in mouse hepatocytes are PI 3-kinase-dependent and rapamycin-sensitive.

Androstadienes↗

PubMed1999

Insulin stimulates phosphorylation of the forkhead transcription factor FKHR on serine 253 through a Wortmannin-sensitive pathway.

In the nematode Caenorhabditis elegans, mutations of the insulin/insulin-like growth factor-1 receptor homologue Daf-2 gene cause developmental arrest at the dauer stage. The effect of Daf-2 mutations is counteracted by mutations in the Daf-16 gene, suggesting that Daf-16 is required for signaling by Daf-2. Daf-16 encodes a forkhead transcription factor. Based on sequence similarity, the FKHR genes are the likeliest mammalian Daf-16 homologues. FKHR proteins contain potential sites for phosphorylation by the serine/threonine kinase Akt. Because Akt is phosphorylated in response to insulin and has been implicated in a variety of insulin effects, we investigated whether insulin affects phosphorylation of FKHR. Insulin stimulated phosphorylation of endogenous FKHR and of a recombinant c-Myc/FKHR fusion protein transiently expressed in murine SV40-transformed hepatocytes. The effect of insulin was inhibited by wortmannin treatment, suggesting that PI 3-kinase activity is required for FKHR phosphorylation. Mutation of serine 253, located in a consensus Akt phosphorylation site at the carboxyl-terminal end of the forkhead domain, abolished the effect of insulin on FKHR phosphorylation. In contrast, mutation of two additional Akt phosphorylation sites, at amino acids threonine 24 or serine 316, did not abolish insulin-induced phosphorylation. These data indicate that FKHR may represent a distal effector of insulin action.

Amino Acid Substitution↗

PubMed1999

Crystal structures of thermostable xylose isomerases from Thermus caldophilus and Thermus thermophilus: possible structural determinants of thermostability.

The crystal structures of highly thermostable xylose isomerases from Thermus thermophilus (TthXI) and Thermus caldophilus (TcaXI), both with the optimum reaction temperature of 90 degrees C, have been determined by X-ray crystallography. The model of TcaXI has been refined to an R-factor of 17.8 % for data extending to 2.3 A and that of TthXI to 17.1 % for data extending to 2.2 A. The tetrameric arrangement of subunits characterized by the 222-symmetry and the tertiary fold of each subunit in both TcaXI and TthXI are basically the same as in other xylose isomerases. Each monomer is composed of two domains. Domain I (residues 1 to 321) folds into the (beta/alpha)8-barrel. Domain II (residues 322 to 387), lacking beta-strands, makes extensive contacts with domain I of an adjacent subunit. Each monomer of TcaXI contains ten beta-strands, 15 alpha-helices, and six 310-helices, while that of TthXI contains ten beta-strands, 16 alpha-helices, and five 310-helices. Although the electron density does not indicate the presence of bound metal ions in the present models of both TcaXI and TthXI, the active site residues show the conserved structural features. In order to understand the structural basis for thermostability of these enzymes, their structures have been compared with less thermostable XIs from Arthrobacter B3728 and Actinoplanes missouriensis (AXI and AmiXI), with the optimum reaction temperatures of 80 degrees C and 75 degrees C, respectively. Analyses of various factors that may affect protein thermostability indicate that the possible structural determinants of the enhanced thermostability of TcaXI/TthXI over AXI/AmiXI are (i) an increase in ion pairs and ion-pair networks, (ii) a decrease in the large inter-subunit cavities, (iii) a removal of potential deamidation/isoaspartate formation sites, and (iv) a shortened loop.

Aldose-Ketose Isomerases↗

PubMed1999

A thermostable xylose isomerase from Thermus caldophilus: biochemical characterization, crystallization and preliminary X-ray analysis.

A highly thermostable xylose isomerase from Thermus caldophilus has been expressed in Escherichia coli. The purified enzyme has an optimum temperature of 363 K. It has been crystallized at room temperature using ammonium sulfate as a precipitant. The crystal belongs to the orthorhombic space group P212121, with unit-cell parameters a = 84.35, b = 123.60, c = 140.24 A. The presence of one molecule of tetrameric xylose isomerase in the asymmetric unit gives a crystal volume per protein mass (Vm) of 2.1 A3 Da-1 and a solvent content of 41% by volume. The crystals initially showed diffraction to 1.7 A Bragg spacing with synchrotron X-rays, and a set of native data extending to 2.3 A resolution has been collected.

Aldose-Ketose Isomerases↗

PubMed1999

Testosterone sulfotransferase: evidence in the guinea pig that this reaction is carried out by 3 alpha-hydroxysteroid sulfotransferase.

During the course of isolating, characterizing, and cloning estrogen and 3-hydroxysteroid sulfotransferases from the guinea pig adrenal gland, it was noted that cytosolic preparations from this tissue would also sulfonate testosterone. Therefore, we set out to isolate and clone the enzyme that performs this reaction. Testosterone sulfotransferase (TST) was isolated from the guinea pig adrenal by using the standard procedures of ion exchange, affinity, and high-performance liquid chromatography. When purified, TST was examined by liquid-phase nondenaturing isoelectric focusing, it was found that the TST activity profile completely overlapped with the activity profile of the 3alpha-hydroxysteroid sulfotransferase (3alphaHST) isoform, but not the 3beta-hydroxysteroid sulfotransferase (3betaHST) isoform. This finding was further investigated by overexpressing the cDNAs for 3alphaHST and 3betaHST in Escherichia coli and examining the expressed proteins for TST activity. This experiment confirmed that 3alphaHST does indeed function as a TST. In addition, 3alphaHST was also found to sulfonate estradiol but not estrone, a finding that further suggested that 3alphaHST may function as a general 17beta-hydroxysteroid sulfotransferase.

Adrenal Glands↗

PubMed1998

Insulin-like growth factor II (IGF-II) secreted from HepG2 human hepatocellular carcinoma cells shows angiogenic activity.

Hepatocellular carcinoma (HCC) is a typical hypervascular tumor. Since insulin-like growth factor II (IGF-II) has been reported to play a significant role in liver regeneration and hepatocarcinogenesis, we initially examined its angiogenic effect on the chorioallantoic membrane (CAM) of 9-day-old chick embryos. We also investigated whether IGF-II secreted from HepG2 human hepatocellular carcinoma cells induces vascularization using the chick embryo CAM. We found that the concentrated conditioned media (CCM) of HepG2 cell culture induced angiogenesis on the CAM. We also identified IGF-II protein in the CCM from HepG2 cells by Western blot analysis. However, CCM from Chang liver cells, which are normal human liver cells and were free of IGF-II, did not induce angiogenesis in the CAM. These results suggest that IGF-II secreted from hepatocellular carcinoma cells may act as an angiogenic factor for the hypervascularization of HCC.

Allantois↗

PubMed1998

Insulin-like growth factor II induced by hypoxia may contribute to angiogenesis of human hepatocellular carcinoma.

Insulin-like growth factor II (IGF-II) is highly expressed during hepatocarcinogenesis (P. Schirmacher et al., Cancer Res., 52: 2549-2556, 1992; B. C. Park et al., J. Hepatol., 22: 286-294, 1995). However, the mechanism of its enhanced expression is largely unknown. In this study, we show that IGF-II mRNA levels are increased within six h of exposing human hepatoma cell cultures to hypoxia, suggesting that hypoxia may be a strong stimulus for the induction of IGF-II expression in the process of hepatocarcinogenesis. This finding and the fact that hepatocellular carcinoma (HCC) is a typical hypervascular tumor (M. Mise et al., Hepatology, 23: 455-464, 1996) imply that IGF-II may play an important role in the development of neovascularization of HCC. Here we demonstrate that IGF-II substantially increases vascular endothelial growth factor (VEGF) mRNA and protein levels in a time-dependent manner in human hepatoma cells. The induction of VEGF by IGF-II was additively increased by hypoxia. Moreover, the direct angiogenic activity of IGF-II was observed in the quantitative chick chorioallantoic membrane assay (M. Nguyen et al., Microvasc. Res., 47: 31-40, 1994). These data suggest that IGF-II may be a hypoxia-inducible angiogenic factor in HCC.

Animals↗

PubMed1998

Involvement of glucocorticoid receptor in the induction of differentiation by ginsenosides in F9 teratocarcinoma cells.

We have previously reported that ginsenosides Rh1 and Rh2 induced the differentiation of F9 teratocarcinoma stem cells [Lee, Y. N., Lee, H. Y., Chung, H. Y., Kim, S. I., Lee, S. K., Park, B. C. and Kim, K. W., In vitro induction of differentiation by ginsenosides in F9 teratocarcinoma cells. Eur. J. Cancer 1996, 32, 1420-1428.]. Since the chemical structure of Rh1 and Rh2 is very similar to that of dexamethasone, a synthetic glucocorticoid, we investigated whether Rh1 and Rh2 act through the glucocorticoid receptor (GR). Immunocytochemistry showed that Rh1 or Rh2 increased the nuclear translocation of GR in the same manner of dexamethasone. In the gel shift assay, glucocorticoid response element (GRE) binding protein in F9 cells was increased by Rh1 or Rh2. To confirm whether the increased binding protein is GR, we performed the competition assay with unlabeled GRE as a specific competitor. Moreover, supershift assay with the GR antibody showed that the binding proteins are GR. In addition, to confirm the Rh1 or Rh2-induced transactivation of GRE promoter, we cotransfected GR expression vector and GRE-luciferase vector. In the luciferase assay, Rh1 or Rh2 potently induced luciferase activity and this induction was blocked by RU486, a potent GR antagonist. Taken together, we suggest that ginsenosides Rh1 and Rh2 may induce the differentiation of F9 cells by stimulating the nuclear translocation of GR.

Animals↗

PubMed1998

Different protein-binding patterns in the P3 promoter region of the human insulin-like growth factor II gene in the human liver cirrhosis and hepatocellular carcinoma tissues.

The P3 promoter of the human insulin-like growth factor II (IGF-II) is the major IGF-II promoter in fetal liver (FL) and hepatocellular carcinoma (HCC). However, little information is available on the transcriptional factors (TFs) controlling IGF-II gene expression in human liver cirrhosis (LC) and HCC tissues. To evaluate the protein-binding patterns in the P3 promoter region, we performed electromobility shift assay (EMSA) and DNase I footprinting assay using nuclear extracts from human FL, LC and HCC tissues. EMSA showed considerable differences in binding patterns of proteins to P3 promoter region according to different nuclear extracts used in this study. By footprinting assay, eight footprints were observed in extracts. In addition, LC extract showed two specific binding at L1 [-80:+30] and L2 [-126:-80] regions, and HCC showed two specific binding at H1 [-176:-120] and H2 [-210:-177] as well as two liver specific binding (L1 and L2). Footprinting after immunoprecipitation indicates that Egr1, Egr2 and Sp1 could bind to P3 promoter directly, while c-jun and c-fos could not bind to these region directly. Further study is required to determine the function of these proteins.

Adult↗