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Biomedical subjects

H L Peng

Publications and source records attributed to H L Peng.

At least 19 recordsLinked to original sources
PubMed2002

Left cervical aortic arch with aneurysm and obstruction: three-dimensional computed tomographic angiography and magnetic resonance angiographic appearance.

Cervical arch is a rare congenital anomaly presumed to result from persistence of the third aortic arch and regression of the normal fourth arch. Rather rare is cervical aortic arch associated with aneurysm and obstruction, with eight known cases reported. Definitive diagnosis with a noninvasive imaging modality is desirable and very important to prevent the need for disaster intervention. We present two cases of a pulsatile mass in the left supraclavicular region. Three-dimensional computed tomographic angiography and magnetic resonance angiography clearly showed a left-sided cervical aortic arch (Haughton type D) with arch aneurysm and coarctation (pseudocoarctation).

Adolescent↗
PubMed2001

Identification of genes induced in vivo during Klebsiella pneumoniae CG43 infection.

A novel in vivo expression technology (IVET) was performed to identify Klebsiella pneumoniae CG43 genes that are specifically expressed during infection of BALB/c mice. The IVET employed a UDP glucose pyrophosphorylase (galU)-deficient mutant of K. pneumoniae which is incapable of utilizing galactose and synthesizing capsular polysaccharide, as demonstrated by its low virulence to BALB/c mice and a white nonmucoid colony morphology on MacConkey-galactose agar. By using a functional galU gene as the reporter, an IVE promoter could render the galU mutant virulent while maintaining the white nonmucoid colony phenotype. A total of 20 distinct sequences were obtained through the in vivo selection. Five of them have been identified previously as virulence-associated genes in other pathogens, while another five with characterized functions are involved in regulation and transportation of nutrient uptake, biosynthesis of isoprenoids, and protein folding. No known functions have been attributed to the other 10 sequences. We have also demonstrated that 2 of the 20 IVE genes turn on under iron deprivation, whereas the expression of another five genes was found to be activated in the presence of paraquat, a superoxide generator.

Animals↗
PubMed2000

Identification of genes present specifically in a virulent strain of Klebsiella pneumoniae.

Klebsiella pneumoniae is a common cause of septicemia and urinary tract infections. The PCR-supported genomic subtractive hybridization was employed to identify genes specifically present in a virulent strain of K. pneumoniae. Analysis of 25 subtracted DNA clones has revealed 19 distinct nucleotide sequences. Two of the sequences were found to be the genes encoding the transposase of Tn3926 and a capsule polysaccharide exporting enzyme. Three sequences displayed moderate homology with bvgAS, which encodes a two-component signal transduction system in Bordetella pertussis. The rest of the sequences did not exhibit homology with any known genes. The distribution of these novel sequences varied greatly in K. pneumoniae clinical isolates, reflecting the heterogeneous nature of the K. pneumoniae population.

Bacterial Proteins↗
PubMed1999

Breast cancer detection using magnetic resonance imaging in breasts injected with liquid silicone.

Two patients who had received silicone injections in their breasts several years ago presented with breast complaints. Excluding cancer in these patients was very difficult. Mammograms were very difficult to interpret, as were the physical findings. Carcinoma was successfully detected by magnetic resonance imaging. When women who underwent the injudicious injection of silicone reach the cancer-prone age, the examining physicians should have a greater awareness of the detection and management of carcinoma coexistent with silicone mastopathy. We think that MRI is potentially valuable in the evaluation of the breast lesions; it plays an important role in the detection of breast cancer in breasts augmented with liquid silicone.

Breast Implants↗
PubMed1999

Characterization of mdcR, a regulatory gene of the malonate catabolic system in Klebsiella pneumoniae.

The Klebsiella pneumoniae mdcR gene, which encodes a LysR-type regulator, was overexpressed in Escherichia coli. Purified MdcR was found to bind specifically to the control region of either the malonate decarboxylase (mdc) genes or mdcR. We have also demonstrated that MdcR is an activator of the expression of the mdc genes, whereas it represses the transcription of the putative control region of mdcR, PmdcR, indicating a negative autoregulatory control.

Amino Acid Sequence↗
PubMed1999

Characterization of a putative Pseudomonas UDPglucose pyrophosphorylase.

A UDP-glucose pyrophosphorylase encoding gene was identified through functional complementation screening by using an Escherichia coli galU mutant. Sequence analysis of the gene indicated that it is most likely derived from a Pseud monas sp. The gene is located immediately upstream and transcribed in the same direction of the gor (glutathione reductase) gene and is capable of encoding a protein 30,943 daltons in size. The gene product synthesized in Escherichia coli was purified and its biochemical properties characterized. The recombinant UDP-glucose pyrophosphorylase exhibited a molecular weight of 130 kDa, suggesting a tetrameric organization of the gene product. Two mutant forms of the enzyme were identified. The activity of the mutant enzyme with a tyrosine to histidine (Y26 1H) substitution was found to be greatly reduced. On the other hand, the tyrosine to cysteine (Y84C) substitution resulted in an enzyme that functions normally at 37 degrees C but rather poorly at temperatures lower than 30 degrees C.

Amino Acid Sequence↗
PubMed1999

Identification of a novel family of human Rab-like small GTP-binding proteins.

The presence of a novel family of Rab-like proteins (Rlp) in the human genome is reported. The gene encoding the Rlp-2 was isolated from a human lymphocyte genomic library. The Rlp-2 gene is intronless and was mapped to chromosome Xq21.3 using fluorescence in situ hybridization. Several cDNA clones encoding the Rlp-1 were identified in a human hippocampus lambda library. Northern analysis revealed a 2.1-kb transcript of Rlp-1 expressed predominantly in brain, heart and skeletal muscle. The transcript was also observed in all examined regions of the human brain at a similar level. An additional gene, termed Rlp-3, which is highly related to Rlp-1 and Rlp-2, was found in the GenBank Data Base. The predicted molecular mass for Rlp is approximately 31 kDa and is consistent with that of Rlp-1 synthesized in Escherichia coli.

Amino Acid Sequence↗
PubMed1998

Outbreak of Candida albicans fungaemia in a neonatal intensive care unit.

During a 4-month period, 9 premature infants hospitalized in a neonatal intensive care unit (NICU) developed Candida albicans fungaemia. All 9 infants received antifungal agents. Fluconazole was administered in 7 patients and successfully eradicated this organism in 6 with no adverse effects. For epidemiological investigation, 64 environmental specimens and hand-washings of all 54 staff members involved in the NICU were examined for the presence of this organism. No C. albicans could be identified from environmental sources, while the hand-washing of 1 nurse was C. albicans-positive. Two genotyping methods, including electrophoretic karyotyping using contour-clamped homogeneous electric field gel electrophoresis and polymerase chain reaction-based direct sequencing of rRNA gene, were used in the analysis of the isolates recovered from blood cultures of the infants, the hand-washing of the nurse and 7 control isolates. Both methods yielded comparable results and revealed that all 13 isolates from infected infants as well as the isolate from hand washing of the nurse were of the same genotype while the control isolates were distinct. These results suggest that the outbreak of C. albicans fungaemia was caused by a particular strain and possibly via cross-infection. In addition, we showed that fluconazole seemed to be safe and effective in treating C. albicans fungaemia in neonates, although the data were limited.

Antifungal Agents↗
PubMed1998

On the cellular mechanism for the effect of acidosis on vascular tone.

The role of smooth muscle [Ca2+]i and membrane potential for the relaxation to hypercapnic (increased CO2) and normocapnic (unchanged CO2) acidosis is not complete understood. It is often stated that membrane hyperpolarization plays an important role but this has not been vigorously tested. In this study we investigated isolated rat cerebral small arteries under isobaric conditions. Lumen diameter was measured simultaneously with either [Ca2+]i or membrane potential, and acidosis was induced by increasing PCO2 or reducing HCO3- of the bathing solution or by adding HCI to a nominally bicarbonate-free solution. Confocal microscopy verified loading of smooth muscle cells with fluorescent dyes. Acidosis always reduced myogenic tone at transmural pressures between 20 and 120 mmHg. Acidification at a transmural pressure of 40 mmHg caused an increase in diameter and a decrease in [Ca2+]i. This was also seen in the presence of L-NNA and after depolarization with 50 mM K+. The response to hypercapnic and normocapnic acidosis was similar. However, while hypercapnic acidosis caused hyperpolarization, normocapnic acidosis caused depolarization. Dilatation, decrease of [Ca2+]i and depolarization, was also seen with reduction of pH in bicarbonate-free solution. We conclude that the isobaric relaxation to both hypercapnic and normocapnic acidosis is most likely mediated by a reduction of [Ca2+]i. Membrane potential may on the other hand not play a major role for this reduction of [Ca2+]i and it is possible that molecular CO2 has an effect on the membrane potential.

Acidosis↗
PubMed1998

Effect of acidosis on tension and [Ca2+]i in rat cerebral arteries: is there a role for membrane potential?

The cellular mechanism responsible for the reduction of tension in cerebral small arteries to acidosis is not known. In this study the role of smooth muscle intracellular Ca2+ concentration ([Ca2+]i) and membrane potential for the relaxation to acidosis was investigated in isolated rat cerebral small arteries. Isometric force was measured simultaneously with [Ca2+]i (fura 2) or with membrane potential (intracellular microelectrodes), and acidosis was induced by increasing PCO2 or reducing HCO3- of the bathing solution. Both hypercapnic and normocapnic acidosis were associated with a reduction of intracellular pH [measured with 2',7'-bis-(carboxyethyl)-5 (and -6)-carboxyfluorescein], caused relaxation, and reduced [Ca2+]i. However, whereas hypercapnic acidosis caused hyperpolarization, normocapnic acidosis was associated with depolarization. It is concluded that a reduction of [Ca2+]i is in part responsible for the direct effect of the acidosis on the vascular smooth muscle both during normo- and hypercapnia. The mechanism responsible for the reduction of [Ca2+]i differs between the hypercapnic and normocapnic acidosis, being partly explained by hyperpolarization during hypercapnic acidosis, whereas it is seen despite depolarization during normocapnic acidosis.

Acidosis↗
PubMed1998

cDNA cloning and expression analysis of the human UDPglucose dehydrogenase.

A cDNA clone encoding the human UDPglucose dehydrogenase was isolated from a liver cDNA library. The cDNA is 2,355 bp in length with an open reading frame which is capable of encoding a protein of 494 residues. The predicted primary sequence of the gene product is in good agreement with that of the bovine enzyme determined previously found by means of protein sequencing. Two major transcripts of the UDPglucose dehydrogenase gene with sizes of 2.8 and 2.35 kb, respectively, were observed by Northern analysis. The gene was found to be expressed in a variety of tissues with the highest level in liver, consistent with the physiological function of the enzyme in excretion of endo- and xenobiotics compounds.

Amino Acid Sequence↗
PubMed1997

pH and smooth muscle.

In this paper, the control of vascular smooth muscle intracellular pH (pHi) and the mechanisms of importance for the vasodilation to acidosis are reviewed. The three transport pathways of importance for the control of pHi are a sodium-coupled bicarbonate transport, a Na,H-exchanger and a Cl,HCO3- exchange. While the two latter pathways are present in all smooth muscle cells studied, the sodium-coupled bicarbonate transport may be present in two forms which are either coupled to chloride efflux or are independent of chloride. The chloride-independent pathway seems electroneutral, indicating a 1:1 stoichiometry. All three transporters can be activated by vasoactive hormones and the second messengers involved are under intense investigation. With respect to the mechanisms involved in the vasodilation to acidosis, there seems to be a nitric oxide-dependent pathway as well as a direct effect of acidosis on the smooth muscle cells. In some preparations, prostanoids may also be involved. The direct vasodilator effect of acidosis is probably mediated through reduction of extracellular pH and the acidosis is associated with a reduction of the intracellular calcium concentration, which could explain the reduction of smooth muscle tone.

Acidosis↗
PubMed1997

Identification and characterization of acoK, a regulatory gene of the Klebsiella pneumoniae acoABCD operon.

By using transposon insertional mutagenesis and deletion analyses, a recombinant clone containing the region upstream of the acoABCD operon of Klebsiella pneumoniae was found to be required for acetoin-inducible expression of the operon in Escherichia coli. The nucleotide sequence of the region was determined, and it displayed an open reading frame of 2,763 bp that is transcribed divergently to the acoABCD operon. This gene, designated acoK, is capable of encoding a protein with an overall 58.4% amino acid identity with MalT, the transcriptional activator of the E. coli maltose regulon. A conserved sequence for nucleotide binding at the N-terminal region, as well as a helix-turn-helix motif belonging to the LuxR family of transcriptional regulators at the C terminus, was also identified. Primer extension analysis identified two transcription initiation sites, S1 and S2, located 319 and 267 bp, respectively, upstream of the putative start codon of acoK. Several copies of NtrC recognition sequence [CAC-(N11 to N18)-GTG] were found in the promoter regions of both the acoK gene and the acoABCD operon. Acetoin-dependent expression of the acoABCD operon could be restored in the E. coli acoK mutants by supplying a plasmid carrying an intact acoK, suggesting a transactivating function of the gene product. The AcoK protein overproduced in E. coli was approximately 100 kDa, which is in good agreement with the molecular mass deduced from the nucleotide sequence. A specific DNA binding property and an ATPase activity of the purified AcoK were also demonstrated.

Acetoin↗
PubMed1996

An improved assay for UDPglucose pyrophosphorylase and other enzymes that have nucleotide products.

UDPglucose pyrophosphorylase catalyses the interconversion UDPglucose plus pyrophosphate and glucose 1-phosphate plus UTP. Several assay methods for this enzyme have been described but the only one that can be used to investigate the specificity with respect to various UDPsugars is based on coupling to UTP formation. This assay employs phosphoglycerate kinase to catalyse the formation 1,3- bisphosphoglycerate which is then used to oxidise NADH in the presence of glyceraldehyde 3-phosphate dehydrogenase. We have found that the activity of phosphoglycerate kinase towards UTP is low which limits the usefulness of the assay to very low rates, in agreement with the published recommendation of Hansen et al. Here it is shown that the dynamic range of the assay is increased by more than five fold on addition of nucleoside diphosphate kinase and ADP, which convert UTP to the preferred phosphoglycerate kinase substrate, ATP. It is also shown that the improved assay is suitable for enzymes with other nucleotide triphosphate products.

Adenosine Diphosphate↗
PubMed1996

Rate-determining steps in the biosynthesis of glycogen in COS cells.

Consistent with previous results, overexpression of rabbit skeletal muscle glycogen synthase in COS cells did not lead to overaccumulation of glycogen unless activating Ser-->Ala mutations were present at key regulatory phosphorylation sites 2 (Ser7) and 3a (Ser644) in the enzyme. In addition, we found that expression of glycogenin, glycogen branching enzyme, or UDP-glucose pyrophosphorylase alone in COS cells had no effect on the glycogen level. However, coexpression of the hyperactive 2,3a glycogen synthase mutant with either glycogenin or UDP-glucose pyrophosphorylase led to higher glycogen accumulation than that obtained from the expression of glycogen synthase alone. Coexpression of glycogenin with the 2,3a mutant of glycogen synthase led to the appearance of glycogenin with a lower molecular weight suggestive of reduced glucosylation. Increased glycogen synthesis may lead to competition between glycogenin and glycogen synthase for their common substrate UDP-glucose. In summary, we conclude that (i) glycogen synthase is a primary rate-limiting enzyme of glycogen biosynthesis in COS cells, (ii) that phosphorylation of glycogen synthase is regulatory for glycogen accumulation, and (iii) once glycogen synthase is activated, the reaction mediated by UDP-glucose pyrophosphorylase can become rate-determining.

1,4-alpha-Glucan Branching Enzyme↗
PubMed1996

The importance of conserved residues in human liver UDPglucose pyrophosphorylase.

Comparison of the amino acid sequences of five eukaryotic UDPglucose pyrophosphorylases has identified a number of conserved residues that may be important for substrate binding or catalysis. Using the cloned cDNA for the human liver enzyme, we have investigated the role of several of these residues by site-directed mutagenesis. Changing the single conserved cysteine (residue 123) to serine resulted in an active enzyme, as did mutating the single concerned histidine (residue 266) to arginine. The two conserved tryptophans were each altered to serine; W218S is active while W333S is not. In the latter case, the enzyme does not appear to fold correctly, and a similar result was obtained by mutation to lysine at one (residue 391) of the four conserved arginines. The other three arginines are not essential, as judged by the observation that R389H, R422Q and R445H are all active. The kinetic properties of each active mutant were investigated and in most cases were found to be similar to those of wild-type. The most dramatic change is a sevenfold increase in the Km for magnesium pyrophosphate with C123S. Overall, none of these conserved residues appears to be essential for activity, although such a role cannot be ruled out for W333 and R391 where mutation resulted in defective folding.

Animals↗
PubMed1996

Sequence differences between human muscle and liver cDNAs for UDPglucose pyrophosphorylase and kinetic properties of the recombinant enzymes expressed in Escherichia coli.

UDP-Glc pyrophosphorylase (EC 2.7.7.9) catalyses the interconversion of MgUTP plus Glc1P and UDP-Glc plus MgPPi. Complementation of an Escherichia coli strain lacking this activity has allowed isolation of cDNA encoding this enzyme from a human muscle library. Two forms were identified and the nucleotide sequence of each was determined; they were found to differ only in the 5' region and we suggest that these arise from the use of a different first exon in the two transcripts. These nucleotide sequences are different from that of the cDNA which was isolated previously from a human liver library [Peng, H.-L. & Chang, H.-Y. (1993) FEBS Lett. 329, 153-158] and it is proposed that these liver and muscle forms are derived from different genes. The cDNA for muscle form I, muscle form II, the liver form, and the liver form fused to part of the lacZ gene were expressed in Escherichia coli and the kinetic properties of each enzyme were characterised. Muscle form I and the LacZ/liver fusion enzyme exhibit Michaelis-Menten kinetics towards all substrates while muscle form II has a sigmoidal dependence of rate upon the concentration of MgPPi. The liver form shows Michaelis-Menten kinetics towards MgUTP. For the remaining three substrates, complex kinetics were observed involving a combination of sigmoidicity at low substrate concentration and partial inhibition at high substrate concentration.

Amino Acid Sequence↗