Biomedical subjects

J H Chung

Publications and source records attributed to J H Chung.

At least 19 recordsLinked to original sources

PubMed1999

Identification of the genes encoding enzymes involved in the early biosynthetic pathway of pteridines in Synechocystis sp. PCC 6803.

The biosynthetic pathway for the pteridine moiety of cyanopterine, as well as tetrahydrobiopterine, has been investigated in Synechocystis sp. PCC 6803. Open reading frames slr0426, slr1626, slr0078 and sll0330 of the organism putatively encoding GTP cyclohydrolase I, dihydroneopterine aldolase, 6-pyruvoyltetrahydropterine synthase and sepiapterine reductase, respectively, have been cloned into T7-based vectors for expression in Escherichia coli. The recombinant proteins have been purified to homogeneity and demonstrated to possess expected genuine activities except that of sll0330. Our result is the first direct evidence for the functional assignment of the open reading frames in Synechocystis sp. PCC 6803. Furthermore, the 6-pyruvoyltetrahydropterine synthase gene is demonstrated for the first time in prokaryotes. Based on the result, biosynthesis of cyanopterine is discussed.

Alcohol Oxidoreductases↗

PubMed1999

The biological significance of non-enzymatic reaction of menadione with plasma thiols: enhancement of menadione-induced cytotoxicity to platelets by the presence of blood plasma.

To test the hypothesis that the non-enzymatic reaction of quinones with thiols in plasma can generate reactive oxygens (ROS), thereby leading to potentiated cellular toxicity, we have studied the effect of a representative quinone compound, menadione, on plasma isolated from rats. The experimental results are as follows: (1) menadione generated ROS via non-enzymatic reaction with protein thiols in plasma; (2) the presence of plasma increased menadione-induced cytotoxicity to platelets; (3) pretreatment of plasma with a thiol-depleting agent significantly suppressed menadione-induced ROS and cytotoxicity. These results suggest that the non-enzymatic reaction of menadione with plasma thiols could be an important process in quinone-induced cellular toxicity.

Animals↗

PubMed1999

Cloning of mouse sepiapterin reductase gene and characterization of its promoter region.

We have isolated and characterized approximately 5 kb mouse sepiapterin reductase gene (Spr) and a highly homologous pseudogene (Sprp). The authentic Spr gene is present as a single copy in the mouse genome and is composed of three exons containing the entire coding region. The primer extension experiment located the transcription initiation site in a putative pyrimidine-rich Inr element. The promoter region of the Spr gene is embedded within a CpG island. It was shown that the promoter region is devoid of distinctive TATA and CAAT boxes. Transient transfection of a series of 5' deletion derivatives of the Spr promoter showed the sequence between -83 and -51 to be essential for promoter activity. The pseudogene Sprp lacks promoter region and exon 3.

Alcohol Oxidoreductases↗

PubMed1999

A human Cds1-related kinase that functions downstream of ATM protein in the cellular response to DNA damage.

Checkpoints maintain the order and fidelity of the eukaryotic cell cycle, and defects in checkpoints contribute to genetic instability and cancer. Much of our current understanding of checkpoints comes from genetic studies conducted in yeast. In the fission yeast Schizosaccharomyces pombe (Sp), SpRad3 is an essential component of both the DNA damage and DNA replication checkpoints. The SpChk1 and SpCds1 protein kinases function downstream of SpRad3. SpChk1 is an effector of the DNA damage checkpoint and, in the absence of SpCds1, serves an essential function in the DNA replication checkpoint. SpCds1 functions in the DNA replication checkpoint and in the S phase DNA damage checkpoint. Human homologs of both SpRad3 and SpChk1 but not SpCds1 have been identified. Here we report the identification of a human cDNA encoding a protein (designated HuCds1) that shares sequence, structural, and functional similarity to SpCds1. HuCds1 was modified by phosphorylation and activated in response to ionizing radiation. It was also modified in response to hydroxyurea treatment. Functional ATM protein was required for HuCds1 modification after ionizing radiation but not after hydroxyurea treatment. Like its fission yeast counterpart, human Cds1 phosphorylated Cdc25C to promote the binding of 14-3-3 proteins. These findings suggest that the checkpoint function of HuCds1 is conserved in yeast and mammals.

Amino Acid Sequence↗

PubMed1999

Ethanol co-exposure increases lethality of allyl alcohol in male Sprague-Dawley rats.

Since allyl alcohol and ethanol are both metabolized by alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH), ethanol could affect allyl-alcohol induced toxicity under in vivo coexposure conditions. Male Sprague-Dawley rats were treated with ethanol (2 g/kg, i.p.) simultaneously or 2 h before with allyl alcohol (40 mg/kg, i.p.). Coexposure to allyl alcohol and ethanol resulted in neither enhancement nor protection in allyl alcohol-induced hepatotoxicity at 24 h. However, markedly increased lethality was observed under our coexposure conditions. Pretreatment with 4-methylpyrazole (4-MP) to inhibit ADH did not result in increased lethality to allyl alcohol or ethanol alone, but significantly reduced the lethality of the combined treatment. In contrast, ALDH inhibition increased the lethality of allyl alcohol alone as well as that of the combined allyl alcohol and ethanol treatment. Kinetic studies revealed that combined treatment with allyl alcohol and ethanol resulted in higher blood allyl alcohol levels compared to allyl alcohol alone, and these were accompanied by greater lethality. ADH inhibition increased allyl alcohol blood levels significantly when rats were treated with allyl alcohol alone or allyl alcohol plus ethanol, leading to protection against lethality. In contrast, ALDH inhibition did not affect blood allyl alcohol levels, but resulted in increased lethality. These data suggest a possible role for a metabolite of allyl alcohol, acrolein, in the increased lethality of allyl alcohol and ethanol coexposure in rats.

Alcohol Dehydrogenase↗

PubMed1999

Nitric oxide-enhanced excitotoxicity-independent apoptosis of glucose-deprived neurons.

Glucose deprivation has been shown to elicit neuronal death via extracellular glutamate accumulation. Here we report that immunostimulated glial expression of inducible nitric oxide synthase enhances the apoptotic death of glucose-deprived cerebellar granule cells (CGC) via the excitotoxicity-independent pathway. CGC cultures were immunostimulated by interferon-gamma (100 U/ml) and lipopolysaccharides (1 microg/ml) and 2 days later were challenged by glucose deprivation. Neither a 2-h Glucose deprivation nor a 2-day immunostimulation altered the viability of CGC. A 2-day immunostimulation, however, markedly enhanced the apoptotic death of CGC glucose-deprived for 1 h. The increased apoptotic death of glucose-deprived CGC after immunostimulation was mimicked by the nitric oxide (NO) releasing reagent 3-morpholinosydnonimine (200 microM, 30 min) and was partially prevented by the NO synthase (NOS) inhibitor N(G)-nitroarginine. The enhanced apoptotic death was not blocked by the N-methyl-D-aspartate (NMDA) receptor antagonists D-2-amino-5-phosphovalerate (APV) and dizocilpine (MK-801) or the non-NMDA receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX). Moreover, the NO-induced enhanced apoptotic death occurred without a significant increase of the concentration of glutamate in the bathing medium. Our data indicate that immunostimulated glial cells potentiate the apoptotic death of glucose-deprived CGC in part through the expression of inducible NOS but not through NMDA receptor activation. Potentiation of glucose-deprived CGC death by immunostimulated glial cells may be clinically implicated in the tendency of recurrent ischemic insults to be more severe and fatal than an initial ischemic insult.

Animals↗

PubMed1999

Structure-based identification of a novel NTPase from Methanococcus jannaschii.

Almost half of the entire set of predicted genomic products from Methanococcus jannaschii are classified as functionally unknown hypothetical proteins. We present a structure-based identification of the biochemical function of a protein with an as yet unknown function from a M. jannaschii gene, Mj0226. The crystal structure of Mj0226 protein determined at 2.2 A resolution reveals that the protein is a homodimer and each monomer folds into an elongated alpha/beta structure of a new fold family. Comparisons of Mj0226 protein with protein structures in the database, however, indicate that one part of the protein is homologous to some of the nucleotide-binding proteins. Biochemical analysis shows that Mj0226 protein is a novel nucleotide triphosphatase that can efficiently hydrolyze nonstandard nucleotides such as XTP to XMP or ITP to IMP, but not the standard nucleotides, in the presence of Mg2+ or Mn2+ ions.

Acid Anhydride Hydrolases↗

PubMed1999

Pharmacological characterization of beta2-adrenoceptor in PGT-beta mouse pineal gland tumour cells.

1. The adrenoceptor in a mouse pineal gland tumour cell line (PGT-beta) was identified and characterized using pharmacological and physiological approaches. 2. Adrenaline and noradrenaline, adrenoceptor agonists, stimulated cyclic AMP generation in a concentration-dependent manner, but had no effect on inositol 1,4,5-trisphosphate production. Adrenaline was a more potent activator of cyclic AMP generation than noradrenaline, with half maximal-effective concentrations (EC50) seen at 175+/-22 nM and 18+/-2 microM for adrenaline and noradrenaline, respectively. 3. The addition of forskolin synergistically stimulated the adrenaline-mediated cyclic AMP generation in a concentration-dependent manner. 4. The pA2 value for the specific beta2-adrenoceptor antagonist ICI-118,551 (8.7+/-0.4) as an antagonist of the adrenaline-stimulated cyclic AMP generation were 3 units higher than the value for the betaI-adrenoceptor antagonist atenolol (5.6+/-0.3). 5. Treatment of the cells with adrenaline and forskolin evoked a 3 fold increase in the activity of serotonin N-acetyltransferase with the peak occurring 6 h after stimulation. 6. These results suggest the presence of beta2-adrenoceptors in mouse pineal cells and a functional relationship between the adenylyl cyclase system and the regulation of N-acetyltransferase expression.

Adenylyl Cyclases↗

PubMed1999

Effect of topically applied 0.1% dexamethasone on endothelial healing and aqueous composition during the repair process of rabbit corneal alkali wounds.

PURPOSE: The effects of topical dexamethasone on the endothelial healing and the change of aqueous compositions were investigated during the repair process of alkali-wounded rabbit cornea. METHODS: A central corneal alkali wound was produced by a 60 sec application of a 5.5 mm round filter paper soaked in 1N NaOH onto one eye of each rabbit. The eyes subsequently were treated topically with either 0.1% dexamethasone or a balanced salt solution (BSS) 4 times per day for 8 weeks. Endothelial wound morphometry was performed after alizarin red and trypan blue staining. The concentrations of ascorbic acid, glucose, and the ions, Na+, K+, Ca2+ and Mg2+, were measured in the aqueous humor. RESULTS: Endothelial healing in control (alkali-wounded but not treated with dexamethasone) corneas showed a biphasic pattern of healing: an initial short-term healing for the first week and then a late long-term healing following the secondary endothelial breakdown. Topical administration of 0.1% dexamethasone deterred endothelial healing during the early period and prevented the secondary endothelial breakdown. However, the total repair process of endothelium was accelerated by the dexamethasone-treatment. Among the various components of the aqueous humor examined, ascorbic acid seemed the most sensitive to change caused by the alkali injury and dexamethasone treatment. CONCLUSIONS: The present data indicate that dexamethasone may have a therapeutic potential in the management of endothelial healing after corneal alkali injury.

Administration, Topical↗

PubMed1999

Structural characteristics of tenecin 3, an insect antifungal protein.

Tenecin 3, an antifungal protein, previously isolated from the insect Tenebrio molitor, inhibits growth of the fungus Candida albicans. However, the antifungal mechanism and functions of tenecin 3 remain unknown. As an initial step to study the mechanism and functions, physical and structural properties of tenecin 3 were examined by circular dichroism (CD) analysis and 2D nuclear overhauser effect spectroscopy. These analyses suggest that tenecin 3 has a propensity of random structure with very loose turn-like elements. The CD results also indicate that this random structural propensity is not significantly affected by temperature, pH, and by the presence of organic solvents or sodium dodecyl sulfate (SDS) micelles. However, the hydrodynamic studies suggest that tenecin 3 is not in extended form in spite of its random structural feature.

Animals↗

PubMed1999

Shark liver oil-induced lipoid pneumonia in pigs: correlation of thin-section CT and histopathologic findings.

PURPOSE: To evaluate sequential changes in thin-section computed tomographic (CT) findings after inducement of lipoid pneumonia and provide the histopathologic bases of these findings. MATERIALS AND METHODS: Shark liver oil was administered to 12 sites in seven pigs. Thin-section CT scans were obtained within 1 hour and at 1 week, 2 weeks, 4 weeks, 8 weeks, 12 weeks, and 16 weeks after oil administration. Scans were assessed for opacity, distribution, location at the lobular level, extent, and volume of the lesions. The CT number in consolidation areas also was measured. Findings at CT were correlated with those in the histopathologic specimens. RESULTS: Diffuse ground-glass opacity was noted on all immediately obtained scans. The opacity of the lesions was highest at 1 week; then it decreased gradually to an area of ground-glass opacity. The extent and volume of the lesions decreased at follow-up CT. Histopathologically, the lesions showed a lobular distribution sharply demarcated from the normal lungs. The lobules of decreased volume showed residual thickening of the alveolar walls with bronchiolectasis and mild collagen deposition of the interlobular septa. Pathologic examination of the low-attenuating consolidation area at CT revealed evidence of partial aeration. CONCLUSION: Thin-section CT findings of lipoid pneumonia include ground-glass opacity and airspace consolidation, followed by complete or incomplete resolution with volume loss and septal thickening. Low-attenuating consolidation at CT does not always indicate the presence of fat.

Animals↗

PubMed1999

Fas is expressed in murine pancreatic islet cells and an insulinoma cell line but does not mediate their apoptosis in vitro.

CD4+ lymphocytes are the most important effector cells in autoimmune diabetes of NOD mice, although some role of CD8+ T cells has been demonstrated. However, it is unknown how CD4+ lymphocytes are able to destroy pancreatic beta-cells that do not express MHC (major histocompatibility complex) class II molecules. Apoptotic cell death mediated by an interaction of Fas with Fas ligand (FasL) could be a mechanism by which MHC class II-negative pancreatic beta-cells are destroyed by CD4+ T lymphocytes. We have examined the expression of Fas molecules in pancreatic islet cells, as well as in a NOD-derived mouse insulinoma cell line (MIN6N8). In addition, the role of Fas-mediated apoptosis in pancreatic islet cell death was explored in vitro. Although Fas expression was not detected by flow cytometric analysis, Fas transcripts were demonstrated in MIN6N8 cells and pancreatic islet cells by the sequencing analysis of the cloned reverse transcription polymerase chain reaction products using Fas-specific primers. IFN (interferon)-gamma, tumor necrosis factor-alpha, interleukin-1 and their combinations failed to enhance Fas expression. Unsorted activated splenocytes from diabetic NOD mice had cytotoxic T lymphocyte activity of a small degree against IFN-gamma-treated MIN6N8 cells with FasL upregulation. However, agonistic anti-Fas antibody with or without cycloheximide did not exert cytotoxicity against MIN6N8 cells or pancreatic islets. FasL transfectant cells also did not kill MIN6N8 cells. Our data indicate that pancreatic beta-islet cells express a small amount of Fas molecules but Fas molecules do not mediate apoptosis of islet cells at least in vitro.

Animals↗

PubMed1999

A pathologic study of abdominal lymphangiomas.

Abdominal lymphangiomas are uncommon angiomatous tumor occurring mainly in childhood. This is a retrospective clinicopathologic study of 17 cases of abdominal lymphangioma. The patients included are five children and 12 adults, with a mean age at initial presentation of 30.7 years (age ranges 3-63). The locations of the tumors were mesentery (5), retroperitoneum (4), colon (3), omentum (3), mesocolon (1) and gallbladder (1). Infiltrative growth was more common pattern than entirely circumscribed pattern. Masses were mostly multilocular cysts and contained chyle or serous fluid. On immunohistochemical staining, 16 cases were reactive for either CD31 or factor VIII-related antigen. These fact would suggest that intra-abdominal lymphangiomas simulate the immunohistochemical features of collecting lymphatics. Follow up was possible in 12 cases for 3-50 months (mean 19 months) and only one patient showed local recurrence. Although abdominal lymphangiomas are rare in adulthood and correct preoperative diagnosis is difficult, awareness of such a possibility in adulthood will contribute to make a correct preoperative diagnosis.

Abdominal Neoplasms↗

PubMed1999

Solid mesenchymal hamartoma of the liver in adult.

This paper presents an unusual solid mesenchymal hamartoma of the liver (MHL) in adult. A well defined solid mass in the left lobe of the liver was found in a 57-year-old female. Preoperative radiologic examinations demonstrated solid mass with multifocal calcifications abutting the gallbladder. By light microscopy, the lesion was composed of dense fibrous stroma with hyalinization, bile ducts and thick-walled vessels without hepatocytes. The solid and hyalinized mesenchymal component would suggest an unusual degenerative change representing a burnt-out MHL.

Female↗

PubMed1998

Bacterial expression of tenecin 3, an insect antifungal protein isolated from Tenebrio molitor, and its efficient purification.

Tenecin 3, an antifungal protein isolated from the insect Tenebrio molitor larvae, inhibits growth of the fungus Candida albicans. However, the antifungal mechanism and functions of tenecin 3 have not yet been studied due to its very low availability from the natural source. Here we report an expression system of the recombinant tenecin 3 in E. coli, whose amino acid composition is the same with that of the natural tenecin 3. We also devised a simple and easy procedure to isolate the recombinant protein from the bacterial cell extracts. The recombinant tenecin 3 showed an antifungal activity against C. albicans as the natural tenecin 3 did. Therefore large quantities of tenecin 3 can be easily obtained by the expression and purification system of tenecin 3 described in this report.

Amino Acid Sequence↗

PubMed1998

Immunohistochemical localization of surfactant apoproteins in usual interstitial pneumonia associated with pulmonary carcinoma.

Surfactant apoproteins A and B (SP-A and SP-B) are antigenic determinants of pulmonary surfactant complexes. The role and functional significance of these proteins are largely unknown and the pattern of expression is probably related to the functional maturation of type II pneumocytes. Differential expression of SP-A and SP-B was reported in the developing human lung but little is known of their expression in the chronic injury. We studied 5 surgical cases of usual interstitial pneumonia (UIP) associated with carcinoma to evaluate the expression of pulmonary surfactant apoproteins. These cases were immunohistochemically examined by the streptavidin-biotin complex method using monoclonal antibodies HS-1 and HS-2 against pulmonary surfactant apoprotein A (SP-A) and B (SP-B), respectively. In UIP, SP-B was expressed strongly in type II pneumocytes and Clara cells but bronchiolar epithelium and metaplastic squamous cell lines in the honeycomb lesion were non-reactive. SP-A showed a similar pattern but much weaker reactivity when compared to that of SP-B. Type II pneumocytes in normal lung tissue exhibited weak immunoreactivity and no difference in the intensity of staining between SP-A and SP-B. Neither carcinomatous area nor metaplastic lining cells at honeycomb lesion show immunoreactivity to SP-A and SP-B. These results suggest that type II pneumocytes in the UIP are functionally immature in their expression of the apoprotein types and the metaplastic squamous cells or neoplastic transformed cells do not have molecular characteristics of type II pneumocytes.

Adult↗

PubMed1998

The relative importance of oxidative stress versus arylation in the mechanism of quinone-induced cytotoxicity to platelets.

Our previous studies demonstrated that menadione is cytotoxic to rat platelets. In an attempt to assess the relative contributions of enzymatic redox cycling versus arylation in menadione-induced cytotoxicity, we have studied three quinones with different mechanisms of action: 2,3-dimethoxy-1,4-naphthoquinone (DMNQ; pure redox cycler), menadione (both redox cycler and arylator), and 1,4-benzoquinone (BQ; pure arylator). BQ was more toxic to rat platelets than menadione, while DMNQ did not cause LDH leakage at all. Cellular uptake kinetics revealed that DMNQ concentration taken up by the cells was equivalent to that decreased in incubation medium. On the other hand, the concentrations of BQ and menadione taken into the cells were significantly lower than the decreases in concentrations seen in the incubation medium. This suggests indirectly that BQ and menadione may have undergone arylation, binding to glutathione (GSH) or protein thiols. The difference in arylation capacity between BQ and menadione was well correlated with their relative cytotoxicity (LDH leakage) observed in platelets. All three quinones caused a rapid, extensive depletion of intracellular GSH in platelets. Treatments with BQ and menadione did not result in formation of oxidized glutathione (GSSG), whereas DMNQ showed a time-dependent increase in GSSG. Altogether, these results suggest that enzymatic redox cycling does not play a critical role in quinone-induced cytotoxicity in rat platelets, while arylation is likely to be quinone's primary mechanism of action.

Alkylation↗

PubMed1998

Increased expression of the gentamicin resistance gene by a Tn3 sequence located at the upstream region.

Recombinant plasmids designated as pCH1 and pCH21 were constructed to determine the influence of the bla sequence on the expression of the gentamicin resistance gene which is located downstream of Tn3. Minimal inhibitory concentration (MIC) test and Northern hybridization results with the constructed plasmids showed that the transcription of the gentamicin resistance gene was initiated from the promoter, located downstream from the PstI site within the bla gene, and the hybrid promoter created by IR sequences of Tn3. Although the read-through transcription from the bla promoter did not occur, the transcription of the bla gene increased the expression of the gentamicin resistance gene.

Anti-Bacterial Agents↗