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Biomedical subjects

J H Kwon

Publications and source records attributed to J H Kwon.

At least 19 recordsLinked to original sources

Studies on the development of functional powder from citrus peel.

The suitability of citrus peels, generated as a by-product of the juice industry, as a source of antioxidants was investigated. Citrus peel powder was prepared by lyophilizing 70% ethanol extract from citrus peels. Extraction was carried out at room temperature (20 degrees C) for 72 h. The extract was subjected to gamma-irradiation treatment (20 kGy). The aqueous solutions of citrus peel powder were examined for color characteristics and antioxidant potential in terms of 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging, beta-carotene bleaching and nitrite scavenging activities. There were significant changes in Hunter color values due to irradiation. The a*- and b*-values decreased due to radiation treatment. DPPH radical scavenging, beta-carotene bleaching and nitrite scavenging activities were not affected by irradiation treatment. Nitrite scavenging activity was the highest in the extract at pH 1.2 followed by pH 4.2 and 6.0. These functional properties of the aqueous solution were found to be stable in heat treatment. It could significantly improve oxidative stability of lipids in fish meat system. Based on these results there may be opportunities to use citrus peel powder as a functional component in the food processing industry with gamma irradiation treatment improving its color characteristics without adversely influencing the functional properties.

Antioxidants↗

A pilot study on accelerated sludge degradation by a high-concentration membrane bioreactor coupled with sludge pretreatment.

A new sludge treatment process combining a high MLSS membrane bioreactor with sludge pretreatment techniques was studied in pilot-scale experiments. The membrane bioreactor (MBR) was adopted for high efficiency aerobic digestion. The combination of alkaline-ozone treatment of the mixed liquor in the MBR reactor accelerated the biodegradation process by enhancing biodegradability of the sludge. The hydraulic retention time (HRT) of the reactor was set as 3.1 days and the DO level was 1 mg/L on average. After 5 months of operation, the accumulative total solids reduction was more than 70%. Removal efficiency of volatile solids and non-volatile solids were 76% and 54%, respectively. It was found that a considerable portion of the non-volatile solids was dissolved into ions and then flushed out with the effluent. Also, about 41% and 28% of T-N and T-P in the raw sludge were removed although no biological nutrient removal process was adopted. The experiment was run smoothly without significant membrane fouling, even at the relatively high levels of MLSS concentration (11,000-25,000 mg/L). It is concluded that the newly proposed process can significantly increase the sludge reduction efficiency with much shorter retention times.

Biodegradation, Environmental↗

Acidic and hydrogen peroxide treatment of polyaluminum chloride (PACL) sludge from water treatment.

The water treatment sludge including coagulants cannot be easily removed by conventional dewatering methods. The possibility of hydrogen peroxide (H2O2) oxidation as a pretreatment to enhance the dewaterability of polyaluminum chloride (PACl) sludge from water works was investigated. H2O2 treatment alone was not effective but H2O2 treatment under acidic condition significantly reduced both the cake water content and specific resistance to filtration (SRF), indicating the enhancement of dewaterability and filterability. The filterability after acid/H2O2 treatment was comparable to polymer conditioning and even more dewatered cake than polymer conditioning was produced. By H202 combined with sulfuric acid (H2SO4), leached iron caused Fenton's reaction, which showed a potential to significantly reduce the amount of solids mass and to produce more compact cake with higher filterability.

Aluminum↗

The Vulnerable Attachment Style Questionnaire (VASQ): an interview-based measure of attachment styles that predict depressive disorder.

BACKGROUND: The Vulnerable Attachment Style Questionnaire (VASQ) was developed to provide a brief self-report tool to assess adult attachment style in relation to depression and validated against an existing investigator-based interview (Attachment Style Interview--ASI). This paper describes the development and scoring of the VASQ and its relationship to poor support and major depression. METHOD: Items for the VASQ reflected behaviours, emotions and attitudes relating to attachment relationship style, drawn directly from the ASI. The VASQ was validated against the ASI for 262 community-based subjects. Test-retest was determined on 38 subjects. RESULTS: Factor analysis derived two factors, labelled 'insecurity' and 'proximity-seeking'. The VASQ insecurity dimension had highest mean scores for those with interview-based Angry-dismissive and Fearful styles and was significantly correlated with degree of interview-based insecurity. The proximity-seeking VASQ scores had highest mean for those with Enmeshed interview attachment style and was uncorrelated with ASI insecurity. VASQ scores were highly correlated with a well-known self-report measure of insecure attachment (Relationship Questionnaire) and text-retest reliability of the VASQ was satisfactory. The total VASQ score and the insecurity subscale proved highly related to poor support and to depressive disorder. This was not the case for the proximity-seeking subscale. CONCLUSION: The VASQ is a brief self-report measure that distinguishes individuals with attachment styles vulnerable for depressive disorder. The use of the measure for screening in research and clinical contexts is discussed.

Aged↗

Colonic epithelial cells are a major site of macrophage inflammatory protein 3alpha (MIP-3alpha) production in normal colon and inflammatory bowel disease.

BACKGROUND AND AIM: Macrophage inflammatory protein 3alpha (MIP-3alpha) is a recently described lymphocyte directed C-C chemokine expressed predominately at extralymphoid sites, including the intestine. The aim of this study was to determine whether colonic epithelial cells produce MIP-3alpha and whether its expression is upregulated in inflammatory bowel disease. METHODS AND RESULTS: We found that interleukin 1beta and tumour necrosis factor alpha dose dependently stimulated MIP-3alpha production in Caco-2 and HT-29 intestinal epithelial cells. In cytokine treated Caco-2 and HT-29 cells, a significant increase in MIP-3alpha protein production was observed after three hours and continued for at least 24 hours. Analysis of colonic tissues by quantitative real time polymerase chain reaction and ELISA revealed significantly elevated MIP-3alpha mRNA levels (7.9-fold; p<0.05) and protein levels (8.9-fold; p<0.05) in Crohn's disease compared with controls or ulcerative colitis. MIP-3alpha immunoreactivity in normal colon and inflammatory bowel disease was principally associated with crypt and surface epithelial cells. Moreover, MIP-3alpha protein levels were elevated in primary epithelial cells isolated from patients with inflammatory bowel disease. CONCLUSIONS: These findings indicate that increased enterocyte MIP-3alpha production may play an important role in lymphocyte activation and recruitment to the colonic epithelium in Crohn's disease and ulcerative colitis.

Biomarkers↗

ZBP-89, Sp1, and nuclear factor-kappa B regulate epithelial neutrophil-activating peptide-78 gene expression in Caco-2 human colonic epithelial cells.

We reported previously that human colonic epithelial cells produce the C-X-C chemokine epithelial neutrophil-activating peptide-78 (ENA-78) and that its expression is up-regulated in ulcerative colitis. The aim of this study was to investigate the transcriptional regulation of ENA-78 gene expression in Caco-2 intestinal epithelial cells. Reporter gene transfection and electrophoretic mobility shift assay studies demonstrated that cooperation between two regions of the ENA-78 promoter were required for maximal gene expression in interleukin-1beta-stimulated Caco-2 cells. Binding of activated p50/p65 nuclear factor-kappaB to nucleotides -82 to -91 was essential for interleukin-1beta-dependent gene transcription, whereas binding of constitutively expressed zinc-requiring nuclear factors to nucleotides -125 to -134 (site A) was required for basal gene expression. Scanning mutagenesis of site A demonstrated overlapping binding elements at this locus. One site (CTCCCCC) bound Sp1 and Sp3, and overexpression of Sp1 (but not Sp3) up-regulated basal ENA-78 transcription. Another site (CCCCTCCCCC) was found to bind the zinc finger nuclear factor ZBP-89, and overexpression of this protein significantly repressed ENA-78 reporter gene activity. This study demonstrates that ENA-78 gene expression in Caco-2 intestinal epithelial cells is subject to complex regulation involving the coordinate binding of ZBP-89, Sp1, and nuclear factor-kappaB to the ENA-78 promoter.

Animals↗

A novel recombinant basic fibroblast growth factor and its secretion.

Basic fibroblast growth factor (FGF-2) is a pleiotropic mitogen which plays an important role in cell growth, differentiation, migration, and survival in different cells and organ systems. Recently, several clinical applications for FGF-2 gene transfer are being evaluated in wound healing and collateral artery development to relieve myocardial and peripheral ischemia due to the ability of FGF-2 to regulate the growth and function of vascular cells. However, FGF-2 lacks a classical hydrophobic secretion signal peptide, the FGF-2 chimeras containing various signal sequences have been explored. In this study, a novel recombinant 4sFGF-2 was constructed by replacing nine residues from the amino-terminus of native FGF-2 (Met1 to Leu9) with eight amino acid residues of signal peptide of FGF-4 (Met1 to Ala8) to better increase the secretion level of FGF-2. When the recombinant FGF-2 gene, cloned into the expression vector with CMV promoter, was expressed in COS-7 cells, the recombinant 4sFGF-2 was highly secreted into the media. The secreted 4sFGF-2 showed the same biological activity as the native FGF-2 in the dose-response effects on DNA synthesis and cell growth of rat aortic smooth muscle cells (RASMCs) and NIH3T3 cells. The 4sFGF-2 also was able to activate MAPK as wild FGF-2 in RASMCs. These results indicate that a novel recombinant 4sFGF-2 may be useful as clinical applicability of angiogenic growth factor gene transfer.

3T3 Cells↗

Effects of mixing intensity on tissue-engineered cartilage.

Mechanical forces regulate the structure and function of many tissues in vivo; recent results indicate that the mechanical environment can decisively influence the development of engineered tissues cultured in vitro. To investigate the effects of the hydrodynamic environment on tissue-engineered cartilage, primary bovine calf chondrocytes were seeded on fibrous polyglycolic acid meshes and cultured in spinner flasks either statically or at one of nine different turbulent mixing intensities. In medium from unmixed flasks, CO(2) accumulated and O(2) was depleted, whereas in medium from mixed flasks the concentrations of both gases approached their equilibrium values. Relative to constructs exposed to nonmixed conditions, constructs exposed to mixing contained higher fractions of collagen, synthesized and released more GAG, but contained lower fractions of GAG. Across the wide range of mixing intensities investigated, the presence or absence of mixing, but not the intensity of the mixing, was the primary determinant of the GAG and collagen content in the constructs. The all-or-none nature of these responses may provide insight into the mechanism(s) by which engineered cartilage perceives changes in its hydrodynamic environment and responds by modifying extracellular matrix production and release. 2001 John Wiley & Sons, Inc.

Animals↗

Coupling of cAMP/PKA and MAPK signaling in neuronal cells is dependent on developmental stage.

Neurite formation, an essential feature of neuronal development, is believed to involve participation of the ras-mitogen-activated protein kinase (MAPK) and cAMP-dependent protein kinase A (cAMP/PKA)-mediated signaling pathways. These pathways have been studied extensively in the rat pheochromocytoma cell line PC12, and current hypotheses suggest a single effector mechanism resulting from the convergence of cAMP/PKA and MAPK signaling. However, based on observations using a central neuronal progenitor cell line, AS583-8, there also exists evidence that the two signaling pathways may act independently resulting in neurites with differing dynamic features. In the present study, the upstream components of cAMP/PKA signaling were examined in AS583-8 cells as well as possible interactions with the MAPK pathway. We found that activation of PKA is both necessary and sufficient for the elaboration of rapidly forming processes, typical of the cAMP response. In addition, blockade of the MAPK pathway has no effect on the cAMP response, suggesting that activation of the cAMP/PKA pathway can stimulate neurite formation independent of the MAPK pathway. In order to evaluate which cell line model, PC12 vs AS583-8, best reflects the signaling features of developing central neurons, we examined interactions between cAMP/PKA and MAPK signaling in primary neuronal cultures from several brain regions. In immature cultures (1-day-old), at a point where the initiation of neurite formation is maximal, no interaction was observed. In more mature cultures (7 days old), where synaptic contacts have been established, we found a weak but reproducible activation of MAPK following stimulation of the cAMP/PKA pathway. These results suggest that cAMP/PKA and MAPK signaling act independently at the initiation of neuritogenesis but become coupled during later stages of neuronal development. Therefore, the interaction of the two pathways may be cell stage (younger vs older) specific and may participate in cellular functions that take place after initial neurite formation.

Animals↗

Erythropoietin response is inadequate in cancer patients receiving chemotherapy.

The level of serum erythropoietin (EPO) is inappropriately decreased in cancer patients and has been advocated as the main cause of their anemia. In cancer patients, chemotherapy results in a cumulative anemia severe enough to require transfusion. We investigated the changes in serum EPO, hemoglobin, ceruloplasmin, and copper levels in cancer patients receiving chemotherapy. There was a weak but significant inverse relationship between hemoglobin and log[EPO] (r = -0.41; P < .001). Observed/expected serum EPO ratios decline with repeated chemotherapy indicating inadequate EPO response for the degree of anemia. There was no difference in the severity of anemia and in the degree of EPO response between platinum- and non-platinum-treated patients. Ceruloplasmin, copper, and ferritin levels did not change during chemotherapy. Our results suggest that the EPO response is inadequate for the degree of anemia and justifies the use of recombinant human EPO in cancer patients receiving chemotherapy.

Adolescent↗

CDNA cloning of chloroplast omega-3 fatty acid desaturase from Capsicum annuum and its expression upon wounding.

A clone for a plastid omega-3 fatty acid desaturase (FAD) was isolated from a leaf cDNA library of hot pepper (Capsicum annuum L.). The nucleotide sequence of a 1,317 bp open reading frame in the CachFAD showed 80.9% homology with that of tobacco plant. It codes for a polypeptide of 438 amino acids with molecular mass of 50.5 kDa and a pI of 8.14. The CachFAD had a putative transit peptide for targeting the chloroplast. Genomic Southern hybridization indicated that it exists as small gene family. Northern hybridization revealed that its mRNA was present in leaves, but not in roots. Transcript levels in the leaves upon wounding increased rapidly to reach the first peak between 1-3 h, decreased thereafter and slightly increased at 24 h after wounding. The levels of linolenic acid (18:3) in wounded leaves also reached the first peak at 6 h, decreased thereafter and reached the second peak at 18 h. These results indicated that wounding not only enhanced the accumulation of the CachFAD mRNA but also increased the conversion of linoleic acid (18:2) to linolenic acid (18:3) in leaf lipids of hot pepper.

Amino Acid Sequence↗

Intrathoracic migration of Steinmann pins.

The migration of surgical wires and pins placed for repair of orthopedic injury is well recognized. Such migration usually follows a retrograde path and the wires protrude near their entry point into the native bone. Occasionally, the migration occurs in an antegrade manner and produces injury. We describe a case where three Steinmann pins placed for fixation of a humeral neck fracture migrated, one slipping backwards towards the humeral entry point and two pins migrating into the thorax.

Bone Nails↗

Cloning of the histidine biosynthetic genes from Corynebacterium glutamicum: organization and analysis of the hisG and hisE genes.

The physically linked hisG and hisE genes, encoding for ATP-phosphoribosyltransferase and phosphoribosyl-ATP-pyrophosphohydrolase were isolated from the Corynebacterium glutamicum gene library by complementation of Escherichia coli histidine auxotrophs. They are two of the nine genes that participate in the histidine biosynthetic pathway. Molecular genetics and sequencing analysis of the cloned 9-kb insert DNA showed that it carries the hisG and hisE genes. In combining this result with our previous report, we propose that all histidine biosynthetic genes are separated on the genome by three unlinked loci. The coding regions of the hisG and hisE genes are 279 and 87 amino acids in length with a predicted size of about 30 and 10 kDa, respectively. Computer analysis revealed that the amino acid sequences of the hisG and hisE gene products were similar to those of other bacteria.

ATP Phosphoribosyltransferase↗

HIV-1 Nef plays an essential role in two independent processes in CD4 down-regulation: dissociation of the CD4-p56(lck) complex and targeting of CD4 to lysosomes.

Human immunodeficiency virus type 1 (HIV-1) Nef down-regulates CD4 by triggering rapid endocytosis of cell surface CD4. To better understand how Nef induces CD4 down-regulation, we generated a series of Nef mutants with small in-frame deletions in the coding region. Three classes of mutants were obtained. The first class produces neither CD4 down-regulation nor dissociation of the CD4-p56(lck) complex. The second class induces CD4 down-regulation in cells lacking p56(lck) expression, but not in cells with p56(lck);these mutants fail to dissociate CD4 from p56lck. These results show that Nef-mediated CD4 dissociation from p56(lck) is important for CD4 down-regulation. The third class of mutants is able to dissociate the CD4-p56(lck) complex but fails to down-regulate surface CD4; internalized CD4 molecules are recycled back to the cell surface. This result suggests that Nef diverts the CD4 recycling pathway to a degradative pathway. We also demonstrate that Nef associates with phosphatidylinositol-3-kinase (PI3K) activity, which is known to be involved in several aspects of membrane trafficking. However, Nef mutants that cause internalized CD4 to be recycled do not associate with PI3K activity; thus Nef-associated PI3K activity might be involved in the latter process of targeting CD4 to a degradative pathway. We conclude that HIV-1 Nef plays a critical role in multiple processes in CD4 down-regulation: (i) disrupting the CD4-p56(lck) complex on the cell surface to allow CD4 internalization and (ii) diverting the internalized CD4 to a lysosomal pathway for its degradation, likely through a PI3K activity.

CD4 Antigens↗

Myrothecium verrucaria bilirubin oxidase and its mutants for potential copper ligands.

Bilirubin oxidase (EC:1.3.3.5) purified from a culture medium of Myrothecium verrucaria MT-1 (authentic enzyme) catalyzes the oxidation of bilirubin to biliverdin in vitro and recombinant enzyme (wild type) was obtained by using an overexpression system of the bilirubin oxidase gene with Aspergillus oryzae harboring an expression vector. The absorption and ESR spectra showed that both bilirubin oxidases are multicopper oxidases containing type 1, type 2, and type 3 coppers similar to laccase, ascorbate oxidase, and ceruloplasmin. Site-directed mutagenesis has been performed for the possible ligands of each type of copper. In some mutants, Cys457 --> Val, Ala, His94 --> Val, and His134.136 --> Val, type 1 and type 2 copper centers were perturbed completely and the enzyme activity was completely lost. Differing from the holoenzyme, these mutants showed type 3 copper signals. However, the optical and magnetic properties characteristic of type 1 copper were retained even by mutating one of the type 1 copper ligands, i.e., a mutant, Met467 --> Gly, showed a weak but apparent enzyme activity. A double mutant His456.458 --> Val had only type 1 Cu, showing a blue band at 600 nm (epsilon = 1.6 x 10(3)) and an ESR signal with very narrow hyperfine splitting (A parallel = 7.2 x 10(-)3 cm-1). Since the type 2 and type 3 coppers are not present, the mutant did not show enzyme activity. These results strongly imply that the peculiar sequence in bilirubin oxidase, His456-Cys457-His458, forms an intramolecular electron-transfer pathway between the type 1 copper site and the trinuclear center composed of the type 2 and type 3 copper sites.

Amino Acid Sequence↗

Site-directed mutagenesis of a possible type 1 copper ligand of bilirubin oxidase; a Met467Gln mutant shows stellacyanin-like properties.

In our previous paper, we reported a mutant of recombinant Myrothecium verrucaria bilirubin oxidase, in which the Met467 residue was replaced by Gly [Shimizu, A. et al. (1999) Biochemistry 38, 3034-3042]. This mutant displayed a remarkable reduction in enzymatic activity and an evident decrease in the intensity of the absorption band around 600 nm (type 1 charge transfer transition). In this study, we report the preparation of three Met467 mutants (Met467Gln, Met467His, and Met467Arg) and characterize their enzymatic activities, midpoint potentials, and absorption and ESR spectra. Met467His and Met467Arg show no enzymatic activity and a great reduction in the intensity of the absorption band around 600 nm. Furthermore, their ESR spectra show no type 1 copper signal, but only a type 2 copper signal; however, oxidation by ferricyanide caused the type 1 copper signal to appear. On the other hand, Met467Gln as expressed shows both type 1 and type 2 copper signals in its ESR spectrum, the type 1 copper atom parameters being very different from usual blue copper proteins but very similar to those of stellacyanin. The enzymatic activity of the Met467Gln mutant for bilirubin is quite low (0.3%), but the activity for potassium ferrocyanide is similar (130%) to that of the wild type enzyme. These results indicate that Met467 is important for characterizing the features of the type 1 copper of bilirubin oxidase.

Amino Acid Sequence↗

Increased MAPK activity and MKP-1 overexpression in human gastric adenocarcinoma.

Mitogen-activated protein kinase (MAPK) has been known to play a critical role in the regulation of the carcinogenesis in human cancers. In an effort to understand the functional role of the MAPK in the carcinogenesis of human gastric tissues, we examined the changes of MAPK levels in human gastric adenocarcinoma. We found that increased MAPK activity was accompanied by overexpression of mitogen-activated protein kinase phosphatase-1 (MKP-1), suggesting that signaling pathways leading to the activation of MAPK and the induction of MKP-1 expression are associated with carcinogenesis of human gastric adenocarcinoma.

Adenocarcinoma↗

Cloning of the histidine biosynthetic genes of Corynebacterium glutamicum: organization and sequencing analysis of the hisA, impA, and hisF gene cluster.

The hisA and hisF genes of Corynebacterium glutamicum were cloned by transforming histidine auxotrophic Escherichia coli with the genomic DNA library. They are two of the eight genes that participate in the histidine biosynthetic pathway. Cloned DNA fragments containing the genes can also complement hisH and hisI auxotrophs of Escherichia coli, suggesting that the four genes are clustered in the genome. We determined the nucleotide sequences of the minimal fragment containing the hisA and hisF genes, which are separated by the impA gene. The coding regions of the hisA and hisF genes are 245 and 257 amino acids in length with a predicted size of about 26 and 27 kDa, respectively. These are in good agreement with the sizes of proteins expressed in E. coli. A high similarity was observed in comparison of nucleotide sequences of each protein between C. glutamicum and other species, as well as those between hisA and hisF genes of C. glutamicum.

Aldose-Ketose Isomerases↗