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H Pang

Publications and source records attributed to H Pang.

At least 55 records · Page 3Linked to original sources
PubMed1997

Genetic variation of envelope gp90 gene of equine infectious anemia virus isolated from an experimentally infected horse.

Six strains of equine infectious anemia virus (EIAV) were recovered from febrile and non-febrile stages of a horse experimentally infected with the P337-V70 strain given once to a horse. The env gp90 genes of the isolates, the P337-V70 and P337-V26, avirulent virus derived from the P337-V70 strain, were sequenced. A comparison of the gp90 gene sequences revealed that amino acid variations among the viruses tested showed as high as 8.2 to 11.5%. In addition, the comparison also indicated that the isolates that recovered from the non-febrile stage were contained in nucleotide insertions in the principal neutralizing domain (PND) region. The insertions were arranged regularly with smaller segments. The nucleotide sequence of the P337-V26 gp90 gene was found to contain a six-nucleotides insertion and seven nucleotide substitutions outside the PND region, when compared with that of the P337-V70 strain.

Amino Acid Sequence↗
PubMed1996

Lipopolysaccharide of the Helicobacter pylori type strain NCTC 11637 (ATCC 43504): structure of the O antigen chain and core oligosaccharide regions.

Smooth- and rough-form lipopolysaccharides from phenol-water extraction of cells from Helicobacter pylori type strain NCTC 11637 were isolated as the water-soluble component of high-M(r) and water-insoluble low-M(r) gel. Structural investigations were performed on the intact water-soluble smooth-form lipopolysaccharide, various oligosaccharides formed as chemical and enzymic degradation products, and three oligosaccharide fractions liberated by acetic acid hydrolysis from the water-insoluble rough-form lipopolysaccharide. A structure is proposed for the complete polysaccharide component of the smooth-form lipopolysaccharide comprising the O antigen chain, an intervening region, and the inner core oligosaccharide on the basis of 1H and 13C NMR experiments, fast atom bombardment/mass spectrometry, and methylation linkage analysis of permethylated oligo- and polysaccharide derivatives. The most striking feature of the O antigen region in the lipopolysaccharide is the presence of extended chains with fucosylated and nonfucosylated N-acetyllactosamine (LacNAc) units that mimic human cell surface glycoconjugates in normal human granulocytes. The chains are terminated by di- or trimeric Lewis(x) (Le(x)) determinants, which are also found in tumor-associated carbohydrate antigens in many adenocarcinomas.

Carbohydrate Conformation↗
PubMed1996

Molecular characterization of feline immunodeficiency virus genome obtained directly from organs of a naturally infected cat with marked neurological symptoms and encephalitis.

Feline immunodeficiency virus (FIV) was first isolated from cats with immunodeficiency syndrome. Recently, neurological abnormalities and brain lesions were shown in cats infected with FIV. To investigate the FIV genome associated with central nervous system (CNS) lesions, proviral DNA sequences from the V3-V6 region of the FIV env gene were directly amplified from uncultured necropsy tissues of a 2-year-old naturally FIV-infected cat with marked neurological symptoms and encephalitis. By in situ hybridization, FIV RNA was detected mainly in the astrocytes. Fifteen clones isolated from cerebrum, bone marrow and lymph node samples showed only a small number of mutations or deletions in this region. A representative clone, JN-BR1, was distantly related to the previous Japanese strain (TM2) belonging to the subtype B. However, it was relatively close to the Petaluma strain which is known to infect feline brain-derived culture cells and induce brain lesions in inoculated cats. By phylogenetic analysis, the JN-BR1 strain was placed in subtype A that included Petaluma strain and several other American and European strains. The JN-BR1 strain derived from brain with encephalitis in this study and the Petaluma strain may share a common genetic structure that is related to their neuropathogenicity.

Amino Acid Sequence↗
PubMed1996

Semipreparative chromatographic separation of cyclosporin G metabolites generated by microsomes from rabbits treated with rifampicin.

An efficient drug-induced rabbit liver microsomal enzyme system, that may be easily scaled up, has been applied to produce milligram (mg) quantities of cyclosporin G metabolites (CMs). Using hepatic microsomal preparations from rabbits pretreated with rifampicin (RIF) and a NADPH regenerating system, we converted 60% of the cyclosporin G (CsG) to 12 CMs in 2 hr. CMs were recovered by solid-phase extraction. Separations performed on a semipreparative scale with two sequential 250 x 10 mm reversed-phase HPLC columns yielded mg quantities of CMs. The yield of GM1 and GM9 obtained by a single HPLC separation step was estimated to be 1585 +/- 103 micrograms and 648 +/- 33 micrograms, respectively. Other CMs produced by this microsomal enzyme system were GM19, GM4N, GM1c, the aldehydic isomers (GM1cAL and GM1AL), GM14N, GM4N9, GM1A, and GM1cA. The purity of CMs was confirmed by analytical HPLC and their molecular weights (M.W.) were determined by Fast Atomic Bombardment Mass Spectrometry (FABMS). Biotransformation of CsG and cyclosporine (CsA) was also compared using this in vitro model system. Significant difference was found between the rate of aldehydic metabolite formation from CsG and that from CsA. This translates to a faster rate of AM1cAL and AM1AL formation than that of GM1cAL and GM1AL. In addition, the rate of CsG conversion to GM9 appears to be faster than that of CsA conversion to AM9.

Animals↗
PubMed1996

Transcriptional analysis of the 16s rRNA gene in Rickettsia prowazekii.

The control of rRNA synthesis in the etiological agent of epidemic typhus, Rickettsia prowazekii, a slowly growing obligate intracytoplasmic bacterium, was investigated. Transcription of the rickettsial 16S rRNA gene (rrs), of which there is only a single copy, was controlled by a single promoter region, and the site for the initiation of transcription (base A) was found 117 bp upstream of the rrs coding region for the mature product. The promoter region contained an Escherichia coli promoter-like sequence, TTGACA-N17-TATAAC, centered 139 bp upstream of the coding region for the mature product. To investigate whether transcription of the rickettsial rrs responds to amino acid starvation conditions, total RNA was isolated from R. prowazekii-infected mouse L929 cells with or without methionine starvation. The level of newly synthesized 16S rRNA precursors in R. prowazekii, as analyzed by ribonuclease protection assays, decreased significantly after methionine starvation for 6 h and then recovered within 12 h after the addition of methionine. The chemical half-lives of the 16S rRNA precursors in the methionine-starved rickettsiae did not differ significantly from those in the normal rickettsiae. These results suggest that R. prowazekii regulates transcription of the rrs in response to amino acid starvation conditions.

Animals↗
PubMed1996

[Analysis of human hair lead and zinc during various physiological periods].

Hair specimens were collected from 1,477 healthy persons at various physiological periods in different areas of Henan Province and determined for their lead and zinc contents, to study systematically the trend of their changes, correlationship between them and their intrinsic connection. Results showed hair lead level was higher in persons, except neonates and the elderly, living in urban area than in rural one. Hair lead level correlated inversely with hair zinc level in logarithmic scale during various physiological periods, with a coefficient of correlation of 0.971. It suggested there possibly is antagonism between lead and zinc in human bodies. Theoretical explanation of the above results and the possibility to use zinc to prevent and treat the hazard caused by lead were provided.

Adolescent↗
PubMed1995

Lipo-oligosaccharide of Campylobacter lari strain PC 637. Structure of the liberated oligosaccharide and an associated extracellular polysaccharide.

Lipo-oligosaccharide from phenol-water extraction of cells of Campylobacter lari strain PC 637 was separated as a water-insoluble gel of low relative molecular mass (M(r)) from a water-soluble extracellular polysaccharide of high M(r). Structural investigations were performed on the lipo-oligosaccharide and the extracellular polysaccharide, variously using 1H, 13C, and 31P NMR spectroscopy, linkage analysis, and fast atom bombardment-mass spectrometry of permethylated derivatives of the glycans and their products of chemical and enzymic degradation. The following structures are proposed for the highly branched oligosaccharide region: [formula: see text] and for the tetraglycosyl phosphate repeating unit of the extracellular polysaccharide: [formula; see text]

Acetylgalactosamine↗
PubMed1995

Lipo-oligosaccharide of the Campylobacter lari type strain ATCC 35221. Structure of the liberated oligosaccharide and an associated extracellular polysaccharide.

Lipo-oligosaccharide (LOS) from phenol-water extraction of cells of the Campylobacter lari type strain (ATCC 35221) was separated as a water-insoluble gel of low relative molecular mass (M(r)) from a water-soluble extracellular polysaccharide of high M(r). Structural investigations were performed on the liberated oligosaccharide and the extracellular polysaccharide, variously using 1H, 13C, and 31P NMR spectroscopy, linkage analysis, and fast atom bombardment-mass spectrometry of permethylated derivatives of the glycans and their products of chemical and enzymic degradation. The following structures are proposed for the highly branched region of the LOS: [formula: see text] and for the tetraglycosyl phosphate repeating unit of the extracellular polysaccharide: [-(PO3-)-->3)-beta-D-GlcpNAc-(1-->2)-6-d-alpha-L-gul-Hepp -(1-->2)-3-d-beta-D-threo-Penp-(1-->3)-6-d-alpha-L-gul-He pp-]n

Acetylgalactosamine↗
PubMed1995

The citrate synthase-encoding gene of Rickettsia prowazekii is controlled by two promoters.

The transcripts of the citrate synthase-encoding gene (gltA) in Rickettsia prowazekii (Rp), an obligate intracellular parasitic bacterium, were analyzed by RNase protection (RP), primer extension (PE) and in vitro transcription assays. Analysis of the 5' end of the gltA mRNA by RP and PE assays revealed that there were two gltA mRNAs with the 5' ends located at 16 bp and 307 bp upstream from the gltA coding region. Since these two mRNAs might represent two species of mRNA transcribed from two different promoters or a single transcript that was processed to give two mRNAs, an in vitro transcription analysis with purified Rp RNA polymerase (RNAP) was performed to distinguish these two possibilities. Purified Rp RNAP catalyzed the formation of two transcripts initiated from the same nucleotides indicated by RP and PE. Sequence analysis identified Escherichia coli (Ec) promoter-like sequences immediately upstream from both transcription start points (tsp). The first promoter (promoter P1) had the core sequence TTCTAA-N17-TATACT, was 6 bp upstream from the tsp (base A) and was centered at 37 bp upstream from the coding region. The second promoter (promoter P2) had the core sequence ATGAAA-N17-TAAAGT, was 7 bp upstream from the tsp (base T) and was centered at 329 bp upstream from the coding region. This is the first demonstration of multiple promoters in this obligate intracellular parasite which has implications concerning transcriptional regulation.

Amino Acid Sequence↗
PubMed1995

The isolation and characterization of four myelin basic proteins from the unbound fraction during CM52 chromatography.

The unbound fraction from CM52 columns was used as the source of at least four additional myelin basic protein (MBP) molecules. From this fraction we routinely obtained two major fractions called C8-A and C8-B. The C8-A and C8-B fractions were further purified on HPLC. Each contained two proteins in the 17- to 18-kDa range which we called C8-A(H) (higher M(r)), C8-A(L) (lower M(r)), C8-B(H), and C8-B(L). The citrulline values (calculated as citrulline plus ornithine) were high in three of the four proteins, which was accompanied by a compensatory decrease in the arginine values. The compositions clearly identified these four proteins with the citrullinated form of MBP. Western blot analysis showed that both H and L forms reacted with anti MBP antibodies. Partial sequence analysis after cyanogen bromide cleavage, showed that the sequences of both proteins in the C8-B fraction (C8-B(H) and C8-B(L)) were identical to the 18.5-kDa isoform of MBP. Mass spectrometry by electrospray ionization of the C8-B(H) and C8-B(L) provided us with accurate masses of 18,558.08 +/- 8.13 and 17,266.63 +/- 2.24, respectively. We concluded that the H and L proteins from the C8-B fractions were MBPs. Although similar detailed analyses of the C8-A(H) and C8-A(L) have not been done they are also considered to be MBP on the basis of the immunoreactivity with anti MBP antibodies. The origins of these proteins is not known at this time and their functional significance is obscure. The possibility that they are found in early forms of myelin, as components of transitional membranes between oligodendrocytes and myelin or are involved in remyelination, cannot be discounted.

Amino Acid Sequence↗
PubMed1995

Participation of acetylpseudouridine in the synthesis of a peptide bond in vitro.

Uracil, uridine, and pseudouridine were acetylated by refluxing in acetic anhydride, and the products of acetylation were incubated with a synthetic peptide (1-21) that corresponds to the N-terminal 21 amino acid residues of human myelin basic protein. Peptide bond formation, at the N alpha terminus in peptide 1-21, was obtained with acetyluracil and acetylpseudouridine, but not with acetyluridine. Transfer of an acetyl group from acetyluracil and acetylpseudouridine depended on acetylation in the N-heterocycle. X-ray crystallographic analysis definitively established N-1 as the site of acetylation in acetyluracil. Mass spectrometry of the acetylation products showed that one acetyl group was transferred to peptide 1-21, in water, by either acetyluracil or acetylpseudouridine at pH approximately 6. Release of the acetyl group by acylaminopeptidase regenerated peptide 1-21 (mass spectrometry) and automated sequencing (for five cycles) of the regenerated (deacetylated) peptide demonstrated that the N terminus was intact. The findings are discussed in the context of a possible role for pseudouridine in ribosome-catalyzed peptidyltransfer, with particular reference being made to similarities between the possible mechanism of acyl transfer by acetyluracil/pseudouridine and the mechanism of carboxyl transfer by carboxylbiotin in acetyl CoA carboxylase. The possibility that idiosyncratic appearance of a wide range of acyl substituents in myelin basic protein could be related to a peculiar involvement of ribosomal pseudouridine is mentioned.

Acetylation↗
PubMed1995

Chemical structures of the core region of Campylobacter jejuni O:3 lipopolysaccharide and an associated polysaccharide.

The complete structure for the core oligosaccharide region of the water-insoluble low-M(r) lipopolysaccharide of Campylobacter jejuni serotype O:3 from phenol/water extraction of bacterial cells was assigned through studies on derivatives of the liberated oligosaccharide. Structure determinations were performed using 1H-NMR and 31P-NMR spectroscopies, methylation analysis supported by fast-atom-bombardment mass spectrometry, and Smith degradation experiments. It was concluded that the complete chains in the core oligosaccharide had the following structure in which a proportion of the terminal residues were phosphorylated: [formula: see text] From a similar series of experiments, it was concluded that an associated polysaccharide, which was isolated from the water phase of the phenol/water extracts, had the following repeating unit in which a proportion of the previously unknown L-glycero-D-ido-heptose (L-alpha-D-ido-Hep) residues were present as 3-hydroxypropanoyl esters, and were not covalently linked to the lipopolysaccharide: [formula see: text]

Campylobacter jejuni↗
PubMed1995

A rifampicin-induced hepatic microsomal enzyme system for the generation of cyclosporine metabolites.

A drug-induced rabbit hepatic microsomal enzyme system has been developed to produce milligram quantities of cyclosporine metabolites (CMs). Using a rifampicin-induced microsomal preparation in the presence of a NADPH regenerating system, 60% of the cyclosporine (CsA) was converted to CMs in 2 h. The CMs were recovered by solid phase extraction, and separated by gradient high performance liquid chromatography with two Ultrasphere Ocyl (C8) columns connected in tandem. More than 20 CMs were resolved. The quantities of major CMs produced by 45 mg of microsomal proteins were established by comparing peak areas with known concentrations of authentic CM standards. These major CMs included AM1, AM9, AM19, AM4N, AM1c and the aldehydic isomers (AM1cAL plus AM1AL). Other CMs that were not quantified included AM14N, AM4N9, AM1A, AM1c9, and AM1D1. Several CMs remained to be identified. All CMs were detected by radioimmunoassay using a non-specific CsA antiserum. The purity of the CMs were confirmed by fast atomic bombardment mass spectrometry. Similar findings were observed when erythromycin or trolandomycin was used to induce the hepatic microsomal enzymes. The procedure used to generate CMs was simple. With the enzyme fraction derived from one rabbit liver, 90 to 100 mg of CMs can be obtained. In this study, the metabolite patterns of CsA produced by rabbit liver microsomes were shown to resemble those observed for humans. These results indicate the possibility of using rabbit models to predict CsA biotransformation in man. The CMs generated by this enzyme system can be used to acquire information relevant to the situation in man.

Animals↗