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Biomedical subjects

J H Kwak

Publications and source records attributed to J H Kwak.

At least 19 recordsLinked to original sources

Morphological evolution of Ba(NO3)2 supported on alpha-Al2O3(0001): an in situ TEM study.

A key question for the BaO-based NOx storage/reduction catalyst system is the morphological evolution of the catalyst particles during the uptake and release of NOx. Notably, because the formed product during NOx uptake, Ba(NO3)2, requires a lattice expansion from BaO, one can anticipate that significant structural rearrangements are possible during the storage/reduction processes. Associated with the small crystallite size of high-surface area gamma-Al2O3, it is difficult to extract structural and morphological features of Ba(NO3)2 supported on gamma-Al2O3 by any direct imaging method, including transmission electron microscopy. In this work, by choosing a model system of Ba(NO3)2 particles supported on single-crystal alpha-Al2O3, we have investigated the structural and morphological features of Ba(NO3)2 as well as the formation of BaO from Ba(NO3)2 during the thermal release of NOx, using ex-situ and in-situ TEM imaging, electron diffraction, energy dispersive spectroscopy (EDS), and Wulff shape construction. We find that Ba(NO3)2 supported on alpha-Al2O3 possesses a platelet morphology, with the interface and facets being invariably the eight [111] planes. Formation of the platelet structure leads to an enlarged interface area between Ba(NO3)2 and alpha-Al2O3, indicating that the interfacial energy is lower than the Ba(NO3)2 surface free energy. In fact, Wulff shape constructions indicate that the interfacial energy is approximately 1/4 of the [111] surface free energy of Ba(NO3)2. The orientation relationship between Ba(NO3)2 and the alpha-Al2O3 is alpha-Al2O3[0001]//Ba(NO3)2[111] and alpha-Al2O3(1-210)//Ba(NO3)2(110). Thus, the results clearly demonstrate dramatic morphology changes in these materials during NOx release processes. Such changes are expected to have significant consequences for the operation of the practical NOx storage/reduction catalyst technology.

Journal Article↗

Polyoma virus-associated nephropathy and concurrent cytomegalovirus infection in the kidney transplant recipients.

INTRODUCTION: Cytomegalovirus (CMV) and polyoma virus BK (BKV) may both establish latency following primary infection. Frequent reactivation of these viruses can occur in the kidney transplant recipients. BKV may induce CMV gene expression by stimulating cellular regulator proteins or by its own gene regulator proteins. A high rate of concurrent CMV infections has been noted in kidney transplant recipients with polyoma virus-associated nephropathy (PVAN). METHODS: PVAN was identified in 10 of 191 patients who received kidney transplants between October 1998 and September 2003. PVAN was confirmed by allograft kidney biopsy. Four of the 10 patients were complicated by concurrent CMV infection. RESULTS: Two patients had only serological evidence of CMV infection and one patient had CMV gastritis. These three patients were treated with intravenous ganciclovir with good results. Disseminated ganciclovir-resistant CMV disease was demonstrated in the remaining patient. This 34-year-old kidney transplant recipient with PVAN died of multiorgan failure despite antiviral therapy with both ganciclovir and foscarnet. CONCLUSION: PVAN with concurrent CMV infection in kidney transplant recipients showed variable clinical courses including mortality. Further studies are needed to elucidate the influence of PVAN on the pathogenesis of CMV infection.

Adult↗

Protein C and/or S deficiency presenting as peripheral arterial insufficiency.

Although protein C and/or S deficiency has frequently been associated with venous thromboembolic events, instances of arterial thromboses have been reported. However, the exact incidence of protein C and/or S deficiency in patients with peripheral arterial insufficiency has not been established. Furthermore, given the lack of adequate studies to define the natural history and angiographic findings of these patients, the treatment has not been well delineated. Therefore, we conducted a prospective study to investigate the prevalence, characteristic angiographic findings and optimal treatments in patients with peripheral arterial insufficiency associated with protein C and/or S deficiency. Between September 2000 and August 2004, 133 patients who presented with peripheral arterial insufficiency underwent hypercoagulability tests before the initiation of any treatments. Of these, 11 patients (8.3%) with protein C and/or S deficiency were included in this study. There were nine males and two females. The ages ranged from 38 years to 72 years (mean 57 years). All patients showed characteristic angiographic findings: long segment thrombotic occlusion of a main peripheral artery without evidence of atherosclerosis or with mild atherosclerotic changes in the aorta and other major arterial trees. Surgical or endovascular procedures were performed in nine patients: bypass graft in four, thrombectomy in four and catheter-directed thrombolysis in one. Conservative treatment with full anticoagulation was performed in two patients. All patients received pre- and post-operative anticoagulation. Except for one amputated case, clinical and vascular laboratory improvements were achieved in 10 patients. Mean follow-up period was 21 months (range 4-45 months). However, one patient, in whom re-vascularization surgery was performed successfully, discontinued warfarin therapy himself at 10 months after surgery, graft occlusion and limb loss occurred at 30 months after surgery. This initial experience suggests that protein C and/or S deficiency may be an independent risk factor for peripheral arterial insufficiency. Patients who present with peripheral arterial insufficiency and protein C and/or S deficiency demonstrate characteristic angiographic findings. Once the diagnosis of protein C and/or S deficiency is made, patients should be treated with life-long anticoagulation.

Adult↗

A novel plasmid-mediated beta-lactamase that hydrolyzes broad-spectrum cephalosporins in a clinical isolate of Klebsiella pneumoniae.

A new extended-spectrum beta-lactamase with an isoelectric point (pI) of 6.2 was detected in Klebsiella pneumoniae F161 that was isolated from a patient with infection. This strain was highly resistant to the third or fourth generation cephalosporins such as ceftazidime, ceftriaxone, cefoperazone, and cefpirome. Analysis of this strain by the double disk diffusion test showed synergies between amoxicillin-clavulanate (AMX-CA) and cefotaxime, and AMX-CA and aztreonam, which suggested that this strain produced a extended-spectrum beta-lactamase (ESBL). Genetic analysis revealed that the resistance was due to the presence of a 9.4-kb plasmid, designated as pKP161, encoding for new beta-lactamase gene (bla). Sequence analysis showed that a new bla gene of pKP161 differed from bla(TEM-1) by three mutations leading to the following amino acid substitutions: Val84 --> Ile, Ala184 --> Val, and Gly238 --> Ser. These mutations have not been reported previously in the TEM type beta-lactamases produced by clinical strains. The novel beta-lactamase was overexpressed in E. coli and purified by ion exchange chromatography on Q-Sepharose and CM-Sepharose, and then further purified by gel filtration on Sehadex G-200. The catalytic activity of the purified beta-lactamase was confirmed by the nitrocefin disk.

Amino Acid Sequence↗

Milolides, new briarane diterpenoids from the western Pacific octocoral Briareum stechei.

Eleven new briarane-type diterpene lactones, designated milolides (1--8, 10--12), together with four known diterpene lactones (9, 13--15), were isolated from the Micronesian octocoral Briareum stechei collected at Yap, Federated States of Micronesia. One aspect of the stereochemistry of one of the known compounds, solenolide C (9), is revised. Structures of the new compounds were determined from spectral data.

Animals↗

Four new coumarin derivatives from Artemisia keiskeana.

Four new coumarin monoterpene ethers, artekeiskeanols A-D (1-4), and three known coumarins, isofraxidin, fraxidin, and daphnoretin, were isolated from the whole plants of Artemisia keiskeana. All structures were determined from spectral data, and that of artekeiskeanol A (1) was confirmed by synthesis.

Alkaloids↗

Inducible and constitutive expression using new plasmid and integrative expression vectors for Thermus sp.

AIMS: To develop molecular tools and examine inducible and constitutive gene expression in Thermus thermophilus. METHODS AND RESULTS: Two plasmid promoter probe vectors and an integrative promoter probe vector were constructed using a promoterless thermostable kanamycin nucleotidyltransferase (KmR) cassette. Three expression vectors were constructed based on a constitutive promoter J17, that functions in both Thermus and Escherichia coli. An inducible expression vector was constructed using the heat-shock inducible promoter (70 to 85 degrees C) from the dnaK gene of T. flavus, and the malate dehydrogenase gene (mdh) from T. flavus was cloned and expressed in both E. coli and T. thermophilus HB27. CONCLUSION: This report describes the construction and use of improved promoter probe and expression vectors for use in Thermus species. The mdh gene can be used as a high temperature (85 degrees C) reporter gene for Thermus sp. The dnaK promoter is thermo-inducible. SIGNIFICANCE AND IMPACT OF THE STUDY: The expression vectors and molecular tools described here are significant improvements over previously reported vectors for Thermus sp. The mdh gene and the thermo-inducible dnaK promoter will facilitate high temperature studies employing Thermus species.

Escherichia coli Proteins↗

Gene delivery into rat glomerulus using a mesangial cell vector.

To develop an effective protocol of gene transfer into glomeruli, an ex vivo gene delivery system using rat mesangial cells (RMC) as a vector was examined. RMC genetically engineered with a retrovirus harboring the Escherichia coli beta-galactosidase gene was used to estimate the efficacy of gene delivery and the location of the cells within the kidney. The RMC expressing beta-galactosidase, RMCLZ1, was cultured in vitro and the cells were injected into the left kidney through the renal artery of a normal Sprague Dawley rat. At least 1 x 10(6) RMCLZ1 was required for effective gene delivery into glomeruli. One hour and 1, 4, and 14 d after injection, glomeruli were isolated from the left kidneys injected with the cells and the expression of beta-galactosidase in each glomeruli was evaluated. One hour and 1 d after injection, more than 90 and 80%, respectively, of glomeruli from the left kidney showed strong beta-galactosidase activity, while no activity of beta-galactosidase was found in the glomeruli from the right kidneys. The number of glomeruli stained by X-gal and the intensity decreased with time. Fourteen days after injection, about 35% of the glomeruli retained the RMCLZ1. X-gal and periodic acid-Schiff staining of frozen sections obtained 14 d after injection allowed the estimation of the site where the mesangial cells injected were located. The mesangial cells were found mainly in two different locations, the glomerular capillary and the mesangium. The majority (about 90%) of the mesangial cells were located in the glomerular capillary and about 9% of the cells were in the mesangial area. Occasionally, the positive staining was found in proximal tubules and the interlobular artery. Although additional methods are required for the site-specific targeting of the mesangial area, the ex vivo gene transfer to glomeruli is feasible and may be a useful tool for future investigations in the pathological mechanisms of glomerular injury.

Animals↗

Cloning, overexpression and purification of Bacillus subtilis elongation factor Tu in Escherichia coli.

To establish the overexpression and one-step purification system of Bacillus subtilis elongation factor-Tu (EF-Tu), the EF-Tu gene was amplified with or without own ribosome binding site (rbs) by PCR and the only PCR product without rbs was subcloned successfully. For the expression of the EF-Tu gene cloned after PCR amplification, a constitutive expression system and inducible expression system with His6 tag at N-terminus or C-terminus, or glutathione-S-transferase (GST) fusion system were examined in E. coli and B. subtilis. Except GST fusion system in E. coli, however, all other trials were unsuccessful at the step of plasmid construction for the EF-Tu expression. The GST/EF-Tu fusion proteins were highly expressed by IPTG induction and obtained as both soluble and insoluble form. From the soluble GST/EF-Tu fusion protein, EF-Tu was obtained to near homogeneity by one-step purification with glutathione-sepharose affinity column chromatography followed by factor Xa treatment. The purified EF-Tu showed high GDP binding activity. These results indicate that the GST/EF-Tu fusion system is favorable to overexpression and purification of B. subtilis EF-Tu.

Bacillus subtilis↗

Methionyl adenylate analogues as inhibitors of methionyl-tRNA synthetase.

Four stable analogues of methionyl adenylate (3-6) were designed as inhibitors of methionyl-tRNA synthetase and synthesized from 2',3'-isopropylideneadenosine. They strongly inhibited aminoacylation activity of methionyl-tRNA synthetases isolated from Escherichia coli, Mycobacterium tuberculosis, Saccharomyces cerevisiae and human. Among the microorganisms tested, however, these chemicals showed the growth inhibition effect only on E. coli.

Adenosine Monophosphate↗

Methionine analogues as inhibitors of methionyl-tRNA synthetase.

A series of methionine analogues have been synthesized as inhibitors of methionyl-tRNA synthetase and evaluated for their inhibitory activities of E. coli methionyl-tRNA synthetase and bacterial growth. Among them, L-methionine hydroxamate 20 has proved to be the best inhibitor of the enzyme with Ki = 19 microM and showed a growth inhibition against E.coli JM 109, P. vulganis 6059 and C. freundii 8090.

Bacteria↗

Methyloxime-substituted aminopyrrolidine: a new surrogate for 7-basic group of quinolone.

Novel fluoroquinolones containing oxime functionalized aminopyrrolidines have been synthesized. They were found to possess potent antibacterial activities against both Gram-negative and Gram-positive organisms, including methicillin resistant Staphylococcus aureus (MRSA). Among these compounds, LB20277 (compound 12) showed the most favorable in vivo efficacy and pharmacokinetic profile in animals. Based on these promising results, LB20277 was selected as a candidate for further evaluation.

Animals↗

Prolyl endopeptidase inhibitors from caryophylli flos.

Three prolyl endopeptidase inhibitors were isolated and identified as luteolin, quercetin and beta-sitosterol-3-O-beta-D-glucopyranoside with IC50 of 0.17, 0.19 and 27.5 ppm, respectively. The inhibition of two flavonoids were non-competitive with substrate. Twenty authentic flavonoids were tested in order to investigate structure-activity relationship. No significant relationship was found in them, however, catechol moiety of B-ring and 7-OH group in flavonoid skeleton were seemed to be responsible for the stronger activity.

Hydrolysis↗

Purification and characterization of Cop, a protein involved in the copy number control of plasmid pE194.

Cop protein has been overexpressed in Escherichia coli using a T7 RNA polymerase system. Purification to apparent homogeneity was achieved by the sequential chromatography on ion exchange, affinity chromatography, and reverse phase high performance liquid chromatography system. The molecular weight of the purified Cop was estimated as 6.1 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). But the molecular mass of the native state Cop was shown to be 19 kDa by an analytical high performance size exclusion chromatography, suggesting a trimer-like structure in 50 mM Tris-HCl buffer (pH 7.5) containing 100 mM NaCl. Cop protein was calculated to contain 39.1% alpha-helix, 16.8% beta-sheet, 17.4% turn, and 26.8% random structure. The DNA binding property of Cop protein expressed in E. coli was preserved during the expression and purification process. The isoelectric point of Cop was determined to be 9.0. The results of amino acid composition analysis and N-terminal amino acid sequencing of Cop showed that it has the same amino acid composition and N-terminal amino acid sequence as those deduced from its DNA sequence analysis, except for the partial removal of N-terminal methionine residue by methionyl-aminopeptidase in E. coli.

Amino Acid Sequence↗

Isolation and identification of chondroitin sulfates from the mud snail.

Chondroitin sulfates were isolated from the mud snail. For the quantitative analysis of enzymatic digestion products of isolated chondroitin sulfates, strong anion exchange-high performance liquid chromatography (SAX-HPLC) was performed. By the action of chondroitinase ABC, three unsaturated disaccharides 2-acetamide-2-deoxy-3-O-(beta-D-gluco-4-enepyranosyluronic acid)-D-galactose (delta Di-OS), 2-acetamide-2-deoxy-3-O-(beta-D-gluco-4-enepyranosyluronic acid)-6-O-sulfo-D-galactose (delta Di-6S) and 2-acetamide-2-deoxy-3-O-(beta-D-gluco-4-enepyranosyluronic acid)-4-O-sulfo-D-galactose (delta Di-4S) were produced from the mud snail chondroitin sulfates. The analysis showed that relative proportion of delta Di-OS/delta Di-6S/delta Di-4S was 58.7/3.1/38.2. The immunomodulating activity of chondroitin sulfate was examined by cell proliferation assay and these results suggest that it might be a immunosuppressant.

Adjuvants, Immunologic↗

A furan derivative from Cornus officinalis.

Dimethyltetrahydrofuran cis-2,5-dicarboxylate a furan derivative has been isolated from the fruits of Cornus officinalis, and it was isolated for the first time in the nature. The structure was elucidated by NMR spectroscopic techniques.

Dicarboxylic Acids↗

A new prenylated flavonol from the roots of Sophora flavescens.

A new prenylated flavonol, sophoflavescenol (1), together with five known flavonoids, kurarinol, kushenol K, kushenol H, trifolirhizin, and kuraidin, were isolated from the roots of Sophora flavescens. The structure of 1 was determined by spectroscopic analysis. Among the five known flavonoids, kurarinol, kushenol K, and kushenol H showed weak antiviral activity against Herpes simplex virus types I and II.

Antiviral Agents↗