Biomedical subjects

H L Peng

Publications and source records attributed to H L Peng.

32 records · Page 2Linked to original sources

PubMed1996

Virulence and outer membrane properties of a galU mutant of Klebsiella pneumoniae CG43.

Non-mucoid mutants of a Klebsiella pneumoniae wild type strain CG43 were generated by transposon mutagenesis. One of the mutants was incapable of fermenting galactose and was designated CG43-17. Alterations of the bacterial surface including capsule, lipopolysaccharides, and several species of outer membrane proteins were noted. The mutant was avirulent to mice and became highly sensitive to human serum. The defects could not be complemented by the gaIETK operon. Diminished activity of UDP-glucose pyrophosphorylase in CG43-17 suggested that it is a gaIU mutant. This possibility was confirmed because the parental phenotypes could be fully restored in the mutant by transforming it with a human liver cDNA encoding UDP-glucose pyrophosphorylase.

Animals↗

PubMed1996

Identification and characterization of the acoD gene encoding a dihydrolipoamide dehydrogenase of the Klebsiella pneumoniae acetoin dehydrogenase system.

The acoD gene, which encodes a dihydrolipoamide dehydrogenase component of the acetoin dehydrogenase enzyme system of Klebsiella pneumoniae was isolated and the nucleotide sequence determined. The gene is capable of encoding a protein of 465 amino acid residues with conserved binding domains for NAD and FAD, and two redox-active cysteine residues. The acoD gene product exhibited a Michaelis constant of 170 microM for NAD, while NADP can not be used as a substrate. The purified enzyme appeared to be a dimer of the acoD gene product. It did not associate tightly with the E1 and E2 components of either acetoin dehydrogenase or 2-oxoglutarate dehydrogenase to form an active multi-enzyme complex.

Acetoin Dehydrogenase↗

PubMed1996

Anti-lipid peroxidation of gomisin J on liver mitochondria and cultured myocardial cells.

AIM: To study the influences of gomisin J on lipid peroxidation and calcium paradox. METHODS: Using two in vitro models of rat liver mitochondria membrane lipid peroxidation (LPO) and cultured myocardial cells. RESULTS: Gomisin J inhibited Fe2+/ascorbic acid and ADP/NADPH-induced LPO with IC50 (95% confidence limits) 5.5 (4.5-6.7) and 4.7 (2.8-7.8) mumol.L-1, respectively, when cultured myocardial cells preincubated with Ca(2+)-free medium for 2 min were incubated with normal medium containing Ca2+, a marked increase of malondialdehyde (MDA) formation occurred and gomisin J 10 mumol.L-1 protected myocardial cells through decreasing MDA formation. CONCLUSION: Gomisin J inhibits LPO in rat liver mitochondria and protects cultured myocardial cells from being injured by calcium paradox.

Animals↗

PubMed1994

Nucleotide sequence and expression in Escherichia coli of the Klebsiella pneumoniae deaD gene.

The deaD gene of Klebsiella pneumoniae was isolated and its nucleotide sequence determined. The K. pneumoniae gene is highly homologous with the Escherichia coli analog throughout most of the coding region. The deduced primary sequence of the K. pneumoniae deaD gene product is 659 amino acids in length, in contrast with the 571 amino acids of the E. coli deaD product published previously. Sequence comparison revealed several differences near the 3' end of the deaD genes which result in the frame-shift effect. The 3' end sequence of the E. coli deaD gene was therefore analyzed to verify the discrepancy. Our result indicates that the E. coli deaD gene encodes a product of comparable size to the K. pneumoniae DeaD protein, and the carboxyl terminal sequences of the two proteins are highly homologous. In vivo expression of the K. pneumoniae deaD gene in E. coli yielded a 65-kDa protein. Primer extension analysis of the mRNA from K. pneumoniae identified a major transcription start site at an A residue 44 nt upstream of the first in-frame ATG codon.

Amino Acid Sequence↗

PubMed1994

Acetoin catabolic system of Klebsiella pneumoniae CG43: sequence, expression, and organization of the aco operon.

A cosmid clone which was capable of depleting acetoin in vivo was isolated from a library of Klebsiella pneumoniae CG43 cosmids. The smallest functional subclone contained a 3.9-kb DNA fragment of the cosmid clone. Sequencing of the DNA fragment revealed three open reading frames (ORFs A, B, and C) encoding polypeptides of 34, 36, and 52 kDa, respectively. The presence of these proteins was demonstrated by expression of the recombinant DNA clone in Escherichia coli. Considerable similarities between the deduced amino acid sequences of the ORFs and those of the following enzymes were found: acetoin dissimilation enzymes, pyruvate dehydrogenase complex, 2-oxoglutarate dehydrogenase complex, and branched-chain 2-oxo acid dehydrogenase complex of various origins. Activities of these enzymes, including acetoin-dependent dichlorophenolin-dohenol oxidoreductase and dihydrolipoamide acetyltransferase, were detected in the extracts of E. coli harboring the genes encoding products of the three ORFs. Although not required for acetoin depletion in vivo, a possible fourth ORF (ORF D), located 39 nucleotides downstream of ORF C, was also identified. The deduced N-terminal sequence of the ORF D product was highly homologous to the dihydrolipoamide dehydrogenases of several organisms. Primer extension analysis identified the transcriptional start of the operon as an A residue 72 nucleotides upstream of ORF A.

3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide)↗

PubMed1993

Cloning of a human liver UDP-glucose pyrophosphorylase cDNA by complementation of the bacterial galU mutation.

A human liver cDNA clone which encodes the UDP-glucose pyrophosphorylase was isolated by complementation of a bacterial galU mutant. The deduced amino acid sequence of the human enzyme comprised 508 amino acids with a calculated molecular mass of 56,950. The human enzyme significantly resembles those of potato tuber and slime mold with a homology of 46.6% and 43.2%, respectively, in amino acid sequence. No homology was found between the eukaryotic and the prokaryotic enzymes. Northern blotting analysis revealed that the gene was expressed at the highest level in skeletal muscle, followed by liver, heart and kidney.

Amino Acid Sequence↗

PubMed1993

Determination of beta-adrenoceptor subtypes in human pulmonary artery and thoracic aorta by radioligand binding.

The beta-adrenoceptors of human pulmonary artery (PA) and thoracic aorta (TA) were studied by the use of a high specific activity radioligand [125I]pindolol (Pin). To identify the subtypes of beta-adrenoceptors in the 2 blood vessels, the competitive inhibition curves of [125I]Pin by beta 1-antagonist atenolol and beta 2-agonist salbutamol were analyzed using a computer program LIGAND of a mathematical model of the ligand-binding system. The Bmax (15.3 +/- 1.2 fmol/mg protein) and the dissociation constant Kd (44 +/- 4 pmol.L-1) for PA were similar to those for TA (Bmax, 12.8 +/- 1.2 fmol/mg protein, Kd, 45 +/- 4 pmol.L-1). Competitive inhibition analysis showed that the beta 1- and beta 2-adrenoceptor subtypes coexisted in human PA and TA, with beta 1 adrenoceptor dominant. The ratios of beta 1:beta 2 in PA and TA were 2.9:1.0 and 2.1:1.0, respectively.

Albuterol↗

PubMed1992

Cloning, sequencing and heterologous expression of a Klebsiella pneumoniae gene encoding an FAD-independent acetolactate synthase.

The gene encoding the valine-resistant and FAD-independent acetolactate synthase of Klebsiella pneumoniae was isolated and expressed in Escherichia coli. The nucleotide sequence of this gene was determined and it exhibited an open reading frame of 1680 bp in length. In vivo expression of the acetolactate synthase-encoding gene in E. coli revealed a single 60-kDa protein which is consistent with the molecular weight calculated from the deduced amino acid sequence of the gene product. The gene product shares about 20-30% homology with the acetolactate synthases of E. coli, yeast and higher plants.

Acetolactate Synthase↗

PubMed1992

Cloning and expression of the Klebsiella pneumoniae galactose operon.

The entire galactose (gal) operon of Klebsiella pneumoniae was isolated and functionally analyzed in Escherichia coli. The genes encoding galactokinase (galK), galactose-1-phosphate uridyltransferase (galT), and UDP-galactose-4-epimerase (galE) were mapped by complementation analysis. The gene order E-T-K was found to be identical to that of Salmonella spp. and E. coli. Analysis of the nucleotide sequence in the control region revealed significant homology with that of E. coli. Two major sites for transcriptional initiation, both mapped to a cytosyl residue, were identified by primer extension. When the operon is expressed in E. coli, the K. pneumoniae gal gene products make up about 30% of the total cellular proteins. The presence of a powerful promoter responsible for high level synthesis of the gal proteins was also demonstrated using beta-galactosidase as reporter.

Amino Acid Sequence↗

PubMed1991

Molecular epidemiology of Klebsiella pneumoniae.

Plasmid profile, restriction fragment length polymorphism (RFLP) patterned by an rRNA probe, and capsule serotyping of 143 Taiwan local bacteriemic isolates of Klebsiella pneumoniae were analyzed. Our results indicate that K1 and K2 are probably the most prevalent serotypes in Taiwan, accounting for 39% of total isolates tested. A remarkable genotypic heterogeneity was observed among these isolates by the analysis of rRNA gene RFLP patterns and plasmid profile, which highlighted differences that were not discerned by the serological tests used to differentiated these strains. On the basis of these analyses, 27 rRNA gene restriction patterns and 57 plasmid profiles were identified.

Humans↗

PubMed1991

Cloning of GL-7-ACA acylase gene from Pseudomonas sp. 130 and its expression in Escherichia coli.

Using BamHI digested and dephosphorylated pBR322 as vector, a GL-7-ACA acylase gene from Pseudomonas sp. 130 chromosomal DNA was cloned and expressed in Escherichia coli C600. Seven positive clones were detected from 3205 recombinant plasmids with 32P-labeled oligonucleotide probes in situ hybridization. Out of them, three clones which produced active GL-7-ACA acylase were identified by radio-immunological assay, chemical test and chromatographic analysis of reaction mixture. The plasmid DNAs of recombinant pMR5, pMR6 and pMR7 were extracted. Analysis of gel electrophoresis indicated that pMR5 and pMR7 contained the same 6.8kb fragment and the size of the insert in pNR6 was 5.7 kb. The effects of various E. coli hosts on the expression of cloned GL-7-ACA acylase gene is also presented.

Amidohydrolases↗

PubMed1988

Cloning, characterization, and sequencing of an accessory gene regulator (agr) in Staphylococcus aureus.

We have previously identified a gene in Staphylococcus aureus, agr, whose activity is required for high-level post-exponential-phase expression of a series of secreted proteins. In this paper, we describe the cloning of this gene in Escherichia coli by using an inserted transposon (Tn551) as a cloning probe. The cloned gene, consisting of a 241-codon open reading frame containing the site of the transposon insertion, was recloned to an S. aureus vector, pSK265, and shown to be functional in S. aureus. Activity was evaluated by determinations of alpha-hemolysin, beta-hemolysin, and toxic shock syndrome toxin-1 production in early-stationary-phase cultures. The cloned gene showed considerable variation with respect to different exoproteins and different host strains compared with the chromosomal agr determinant; this variation could not be attributed to the higher copy number of the cloned gene and probably reflects inapparent subtleties of the regulatory system.

Amino Acid Sequence↗

PubMedDate not supplied

Outbreak of Candida parapsilosis fungemia in neonatal intensive care units: clinical implications and genotyping analysis.

During a 5-month period, 17 infants hospitalized in neonatal intensive care units of a medical center and a branch hospital developed 18 episodes of Candida parapsilosis fungemia. The mean age at onset was 35 days. Prior to fungemia, all the infants had received hyperalimentation and antibiotics, and 15 infants had had central venous catheters. The presenting symptoms were variable but only vague in 40% of the episodes. Despite administration of antifungal agents, subsequent eradication of fungemia was achieved in only two-thirds of the episodes. None of the environmental samples was positive for C. parapsilosis, while 20% of hand-washing samples of staff working in both units yielded this microorganism. Four genotypes with two main types were identified from 14 outbreak strains and eight genotypes from 14 hand-washing strains, with one type predominant. The results suggest that C. parapsilosis fungemia increases the morbidity and mortality of neonates but does not cause acute lethal events. The outbreak was caused by two main genotypes, possibly via cross-infection by the hands of health care workers.

Candida↗

PubMedDate not supplied

Holocord intramedullary spinal cord astrocytoma: report of one case.

Intramedullary spinal cord astrocytoma in infants is relatively uncommon. Its occurrence is usually confined to the cervical and cervicothoracic regions. In this paper, we report on the case of a 4-month old male infant with low grade holocord intramedullary spinal cord astrocytoma. He had developed progressive weakness of the lower extremities over a month period. Neurological examination revealed flaccid paraplegia as well as complete loss of all modalities of sensation below the T10 level. MRI revealed a large intramedullary mass which was found to be an intramedullary astrocytoma at surgery. This case report presents the clinical features, radiographic findings, and treatment and outcome for this patient together with a review of relevant literature.

Astrocytoma↗