Browse publications by year: 2002

  1. Williams MJ
    Ambio, 2002 Jun;31(4):337-9.
    PMID: 12174604
    MeSH terms: Animals; Artificial Intelligence*; Conservation of Natural Resources; Ecuador; Environment; Fisheries; Ghana; Humans; Population Dynamics*; Technology*; Thailand; Ecosystem*
  2. bin Hussein MZ, Zainal Z, Yahaya AH, Foo DW
    J Control Release, 2002 Aug 21;82(2-3):417-27.
    PMID: 12175754
    Formation of the so-called organic-inorganic nanohybrid material was exploited for the preparation of a controlled release formulation. The inorganic Zn-Al-layered double hydroxide (LDH) was used as a matrix, hosting an active agent or a guest, alpha-naphthaleneacetate (NAA), a plant growth regulator by self-assembly technique. The reverse process, i.e., the deintercalation or release of the guest, NAA was found to be rapid initially, followed by a more sustained release thereafter and this behavior was dependent on the pH of the release medium, the aqueous solution. The mechanism of release has been interpreted on the basis of the ion-exchange process between the NAA anion intercalated in the lamella host and nitrate or hydroxyl anions in the aqueous solution.
    MeSH terms: Aluminum Hydroxide/chemistry*; Delayed-Action Preparations/chemistry; Drug Carriers/chemistry; Hydrogen-Ion Concentration; Hydroxides/chemistry*; Kinetics; Naphthaleneacetic Acids/chemistry*; Plant Growth Regulators/chemistry*; Solutions; Time Factors; X-Ray Diffraction; Spectroscopy, Fourier Transform Infrared; Zinc Compounds/chemistry*; Nanotechnology
  3. Lee J, Wang H, Chia KS, Koh D, Hughes K
    Int J Epidemiol, 2002 Aug;31(4):875-6.
    PMID: 12177037
    MeSH terms: Cause of Death; China/ethnology; Ethnic Groups; Female; Humans; Malaysia/ethnology; Male; Neoplasms/mortality; Neoplasms/epidemiology*; Obesity/complications*; Obesity/epidemiology; Prospective Studies; Risk Factors; Singapore/epidemiology; Body Mass Index; Incidence; Proportional Hazards Models
  4. Ismail BS, Cheah UB, Enoma AO, Lum KY, Malik Z
    Bull Environ Contam Toxicol, 2002 Sep;69(3):444-51.
    PMID: 12177768
    MeSH terms: Agriculture*; Environmental Monitoring; Food Contamination*; Insecticides/analysis; Insecticides/pharmacokinetics*; Organothiophosphorus Compounds/analysis; Organothiophosphorus Compounds/pharmacokinetics*; Vegetables/chemistry*; Ecosystem*
  5. Schurr TG, Wallace DC
    Hum Biol, 2002 Jun;74(3):431-52.
    PMID: 12180765
    In a previous study of Southeast Asian genetic variation, we characterized mitochondrial DNAs (mtDNAs) from six populations through high-resolution restriction fragment length polymorphism (RFLP) analysis. Our analysis revealed that these Southeast Asian populations were genetically similar to each other, suggesting they had a common origin. However, other patterns of population associations also emerged. Haplotypes from a major founding haplogroup in Papua New Guinea were present in Malaysia; the Vietnamese and Malaysian aborigines (Orang Asli) had high frequencies of haplogroup F, which was also seen in most other Southeast Asian populations; and haplogroup B, defined by the Region V 9-base-pair deletion, was present throughout the region. In addition, the Malaysian and Sabah (Borneo) aborigine populations exhibited a number of unique mtDNA clusters that were not observed in other populations. Unfortunately, it has been difficult to compare these patterns of genetic diversity with those shown in subsequent studies of mtDNA variation in Southeast Asian populations because the latter have typically sequenced the first hypervariable segment (HVS-I) of the control region (CR) sequencing rather than used RFLP haplotyping to characterize the mtDNAs present in them. For this reason, we sequenced the HVS-I of Southeast Asian mtDNAs that had previously been subjected to RFLP analysis, and compared the resulting data with published information from other Southeast Asian and Oceanic groups. Our findings reveal broad patterns of mtDNA haplogroup distribution in Southeast Asia that may reflect different population expansion events in this region over the past 50,000-5,000 years.
    MeSH terms: Asia, Southeastern; DNA, Mitochondrial/genetics*; Female; Genetics, Population*; Haplotypes/genetics*; Humans; Male; Polymorphism, Restriction Fragment Length; Genetic Variation*
  6. Kono Y, Yusnita Y, Mohd Ali AR, Maizan M, Sharifah SH, Fauzia O, et al.
    Arch Virol, 2002 Aug;147(8):1623-30.
    PMID: 12181680
    A virus, named Oya virus, was isolated in Vero cell cultures from the lungs of a pig suspected of Nipah virus infection. The virus was revealed as a spherical enveloped RNA virus with a diameter of 79 nm. For identification of Oya virus, RT-PCR was performed. A common primer set for S-RNA of the Simbu serogroup of the genus Bunyavirus was able to amplify a cDNA from Oya virus RNA. The sequence data of the product revealed that the partial gene of Oya virus S-RNA segment had 65-70% homology with published cDNA sequences of Simbu serogroup viruses. The phylogenetic analysis of the data showed that the Oya virus is grouped in Simbu serogroup, but is genetically distinct from the serogroup viruses that have been analyzed molecularly. Serological surveys revealed that the virus distributed widely and densely in Malaysia.
    MeSH terms: Animals; Antibodies, Viral/blood; Base Sequence; Cercopithecus aethiops; Cytopathogenic Effect, Viral; Molecular Sequence Data; Simbu virus/classification; Simbu virus/isolation & purification*; Swine/virology*; Vero Cells; Paramyxovirinae*; Paramyxoviridae Infections/veterinary*; Paramyxoviridae Infections/virology; Reverse Transcriptase Polymerase Chain Reaction
  7. Mirghani ME, Che Man YB, Jinap S, Baharin BS, Bakar J
    Phytochem Anal, 2002 Jul-Aug;13(4):195-201.
    PMID: 12184171
    A simple and rapid Fourier transform infrared (FTIR) spectroscopic method has been developed for the quantitative determination of malondialdehyde as secondary oxidation product in a palm olein system. The FTIR method was based on a sodium chloride transmission cell and utilised a partial least square statistical approach to derive a calibration model. The frequency region combinations that gave good calibration were 2900-2800, and 1800-1600 cm-1. The precision and accuracy, in the range 0-60 mumol malondialdehyde/kg oil, were comparable to those of the modified distillation method with a coefficient of determination (r2) of 0.9891 and standard error of calibration of 1.49. The calibration was cross-validated and produced an r2 of 0.9786 and standard error of prediction of 2.136. The results showed that the FTIR method is versatile, efficient and accurate, and suitable for routine quality control analysis with the result obtainable in about 2 min from a sample of less than 2 mL.
    MeSH terms: Malondialdehyde/analysis*; Oleic Acids/chemistry*; Oxidation-Reduction; Plant Oils/chemistry*; Reproducibility of Results; Spectroscopy, Fourier Transform Infrared/methods*
  8. Tan KH, Nishida R, Toong YC
    J Chem Ecol, 2002 Jun;28(6):1161-72.
    PMID: 12184394 DOI: 10.1023/A:1016277500007
    The major fruit fly attractant component in the floral fragrance of Bulbophyllum cheiri (fruit fly orchid) is methyl eugenol (ME). In the lowland rain forest of Malaysia, the solitary and nonresupinate flowers of the fruit fly orchid attract only males of the ME-sensitive fruit fly species (Bactrocera carambolae, B. papayae. and B. umbrosa. During the morning, the fruit fly orchid flower is visited by many fruit flies, which can sometimes cover the whole flower. The number of visitors dwindles in the afternoon. Headspace analysis of the flower shows a high ME peak in the morning, a small one between 12:00 and 14:00 hr, and no detectable ME peak after 14:00 hr. The process of pollination in the wild is initiated by attraction of fruit flies to floral ME. The flower, with the aid of its specialized hinged see-saw lip (labellum), temporarily traps (< 1 min) a fruit fly pollinator between its lip and column. Just prior to this, the fly is rewarded by the opportunity to feed on the floral attractant found on surfaces of petals, sepals, and lip. The pollinaria borne by two wild B. papayae males (caught on and near the fruit fly orchid flower) are identical in morphology and structure with those obtained from the flower. Many of the B. papayae males (17 of 22 analyzed) attracted to the fruit fly orchid already possessed both ME metabolites, trans-coniferyl alcohol and 2-allyl-4,5-dimethoxyphenol, in their rectal glands. indicating that they had previously consumed ME. In this orchid-fruit fly association, both organisms gain direct reproductive benefits: the orchid flower gets pollinated without having to offer nectar, while the fruit fly boosts its pheromone and defense system, as well as its sexual competitiveness by feeding on the ME produced by the flower.
    MeSH terms: Animals; Diptera/physiology*; Microscopy, Electron, Scanning; Pheromones*; Pollen*; Orchidaceae/physiology*; Orchidaceae/ultrastructure
  9. Zainal M, Ismail SM, Ropilah AR, Elias H, Arumugam G, Alias D, et al.
    Br J Ophthalmol, 2002 Sep;86(9):951-6.
    PMID: 12185113
    BACKGROUND: A national eye survey was conducted in 1996 to determine the prevalence of blindness and low vision and their major causes among the Malaysian population of all ages.

    METHODS: A stratified two stage cluster sampling design was used to randomly select primary and secondary sampling units. Interviews, visual acuity tests, and eye examinations on all individuals in the sampled households were performed. Estimates were weighted by factors adjusting for selection probability, non-response, and sampling coverage.

    RESULTS: The overall response rate was 69% (that is, living quarters response rate was 72.8% and household response rate was 95.1%). The age adjusted prevalence of bilateral blindness and low vision was 0.29% (95% CI 0.19 to 0.39%), and 2.44% (95% CI 2.18 to 2.69%) respectively. Females had a higher age adjusted prevalence of low vision compared to males. There was no significant difference in the prevalence of bilateral low vision and blindness among the four ethnic groups, and urban and rural residents. Cataract was the leading cause of blindness (39%) followed by retinal diseases (24%). Uncorrected refractive errors (48%) and cataract (36%) were the major causes of low vision.

    CONCLUSION: Malaysia has blindness and visual impairment rates that are comparable with other countries in the South East Asia region. However, cataract and uncorrected refractive errors, though readily treatable, are still the leading causes of blindness, suggesting the need for an evaluation on accessibility and availability of eye care services and barriers to eye care utilisation in the country.

    MeSH terms: Adolescent; Adult; Aged; Aged, 80 and over; Blindness/etiology; Blindness/epidemiology*; Cataract/complications; Child; Child, Preschool; Female; Health Surveys; Humans; Infant; Infant, Newborn; Malaysia/epidemiology; Male; Middle Aged; Refractive Errors/complications; Vision, Low/etiology; Vision, Low/epidemiology*; Prevalence; Cluster Analysis; Age Distribution; Sex Distribution
  10. Tan PS, Genc F, Delgado E, Kellum JA, Pinsky MR
    Intensive Care Med, 2002 Aug;28(8):1064-72.
    PMID: 12185426
    We tested the hypothesis that NO contamination of hospital compressed air also improves PaO(2) in patients with acute lung injury (ALI) and following lung transplant (LTx).
    MeSH terms: Air/analysis*; Coronary Artery Bypass; Environment, Controlled; Hospitals, University; Humans; Intensive Care Units; Nitric Oxide/administration & dosage*; Nitric Oxide/adverse effects; Oxygen/analysis*; Pennsylvania; Prospective Studies; Respiration, Artificial/methods*; Respiratory Distress Syndrome, Adult/therapy*; Vasodilator Agents/administration & dosage*; Vasodilator Agents/adverse effects; Lung Transplantation; Treatment Outcome
  11. Chowdhury SM, Omar AR, Aini I, Hair-Bejo M, Jamaluddin AA, Kono Y, et al.
    J. Biochem. Mol. Biol. Biophys., 2002 Aug;6(4):249-55.
    PMID: 12186740
    A study was conducted to isolate and identify chicken anaemia virus (CAV) from field samples of clinically infected broiler chickens in Malaysia. A total of 125 samples were collected from chickens aged 2-6 weeks with clinically depressed and retarded growth, of which five samples were found positive to CAV directly by polymerase chain reaction (PCR). Later, five isolates of CAV from the respective five PCR positive samples were isolated in MDCC-MSB1 cells at passage 4 based on cytopathic effects, PCR and indirect immunofluorescent antibody test. The isolates were identified as BL-1, BL-2, BL-3, BL-4 and BL-5. These CAV isolates were found to resist treatment with chloroform and heat at 37 degrees C for 2 h, 56 degrees C for 30 min and 70 degrees C for 5 min. One of the isolates, BL-5 produced significant reduction (p < 0.001) of hematocrit values (9-19%), pale bone marrow, thymus atrophy and haemorrhages in skin/muscle when inoculated into 1-day old SPF chickens. Restriction enzyme digestion of 926 bp genomic fragments of all the isolates including Cux-1 isolate with HindIII exhibited a similar pattern of bands in 2% agarose gel. The present findings confirmed the presence of CAV in Malaysia.
    MeSH terms: Animals; Antigens, Viral/metabolism; Bone Marrow/virology; Cell Line; Chickens/virology*; Chloroform/pharmacology; DNA Restriction Enzymes/metabolism; Electrophoresis, Agar Gel; Hot Temperature; Malaysia; Microscopy, Fluorescence; Skin/virology; Temperature; Thymus Gland/virology; Time Factors; Polymerase Chain Reaction; Chicken anemia virus/genetics*; Chicken anemia virus/isolation & purification*; Chicken anemia virus/metabolism; Fluorescent Antibody Technique, Indirect
  12. Teo CH, Tan SH, Othman YR, Schwarzacher T
    J. Biochem. Mol. Biol. Biophys., 2002 Jun;6(3):193-201.
    PMID: 12186754
    Ty1-copia-like retrotransposons have been identified and investigated in several plant species. Here, the internal region of the reverse transcriptase (RT) gene of Ty1-copia-like retrotransposons was amplified by PCR from total genomic DNA of 10 varieties of banana. Two to four clones from each variety were sequenced. Extreme heterogeneity in the sequences of Ty1-copia-like retrotransposons from all the varieties was revealed following sequence analysis of the reverse transcriptase (RT) fragments. The size of the individual RT gene fragments varied between 213 and 309 bp. Southern blots of genomic DNA digested from Musa acuminata and other banana varieties probed with W8 clone from M. acuminata and A4 clone from Pisang Abu Nipah showed similar strong, multiple restriction fragments together with other faint hybridization band patterns with variable intensities indicating the presence of many copies of the Ty1-copia-like retrotransposons in the genomes. There was no correlation between retroelement sequence and the banana species (with A or B genomes) from which it arose, suggesting that the probes are not useful for tracking genomes through breeding populations.
    MeSH terms: Base Sequence; Cloning, Molecular; DNA/genetics; Molecular Sequence Data; Phylogeny; Sequence Homology, Nucleic Acid; Blotting, Southern; Polymerase Chain Reaction; Retroelements/genetics*; DNA, Plant; Genome, Plant; Musa/genetics*
  13. Chowdhury SM, Omar AR, Aini I, Hair-Bejo M, Jamaluddin AA, Wan KL, et al.
    J. Biochem. Mol. Biol. Biophys., 2002 Jun;6(3):229-32.
    PMID: 12186760
    Two areas in the chicken anemia virus (CAV) genome have high G:C content with secondary structures. These two G:C rich areas could not be sequenced with Perkin Elmer's Big Dye Terminator Cycle Sequencing Kit. Several modifications were carried out to solve the problem. Finally, a package of modified method was developed to sequence the high G:C areas. The result showed that the Perkin Elmer's Big Dye Terminator Cycle Sequencing Kit with the normal procedures are not suitable for sequencing the high G:C regions of the CAV genome. The present developed method made the Perkin Elmer's Kit useful for the first time to sequence the G:C rich hairpin structures of the CAV genome. The system may be useful to sequence all other G:C rich DNA templates.
    MeSH terms: Animals; Base Sequence; Cloning, Molecular; Cytosine; Genetic Techniques*; Guanine; Models, Genetic; Molecular Sequence Data; Tumor Cells, Cultured; Polymerase Chain Reaction; Genome, Viral*; Sequence Analysis, DNA/methods*; Chicken anemia virus/genetics*
  14. Tarafder MT, Kasbollah A, Saravanan N, Crouse KA, Ali AM, Tin Oo K
    J. Biochem. Mol. Biol. Biophys., 2002 Apr;6(2):85-91.
    PMID: 12186762
    Eight selective nitrogen-sulfur donor ligands have been synthesized from the condensation of S-methyldithiocarbazate (SMDTC) with aldehydes and ketones with a view to evaluating their antimicrobial and cytotoxic activities, and also to correlate the biological properties with the structure of the ligands. The compounds were all characterized by elemental analyses and other physicochemical techniques. SMDTC and the Schiff bases were screened for antimicrobial and cytotoxic activities. SMDTC showed very large inhibition zones (24-44 mm) against bacteria and fungi with a minimum inhibitory concentration (MIC) of 390-25,000 and 1562-6250 microg ml(-1), against different bacteria and fungi, respectively. Streptomycin and nystatin were used as the internal standards against bacteria and fungi, respectively. SMDTC along with its Schiff bases with pyridine-2-carboxaldehyde, acetylacetone and 2,3-butanedione were strongly antifungal and the MIC values were comparable to nystatin. Most of the Schiff bases were strongly cytotoxic. In particular, those with pyridine-2-carboxaldehyde and 2,3-butanedione have CD(50) values of 5.5, 1.9-2.0 microg ml(-1), respectively, against leukemic cells, while against colon cancer cells, the values were 3.7 and 2.0 microg ml(-1), respectively. The glyoxal Schiff base was strongly active only against leukemic cell with CD(50) value of 4.0 microg ml(-1). The present findings have been compared with standard drugs.
    MeSH terms: Anti-Bacterial Agents/chemical synthesis; Anti-Bacterial Agents/pharmacology; Anti-Bacterial Agents/chemistry; Antineoplastic Agents/chemical synthesis; Antineoplastic Agents/pharmacology; Antineoplastic Agents/chemistry; Chemistry, Physical; Drug Screening Assays, Antitumor; Humans; Hydrazines/chemical synthesis; Hydrazines/pharmacology*; Hydrazines/chemistry*; Ligands; Microbial Sensitivity Tests; Schiff Bases/chemical synthesis; Schiff Bases/pharmacology; Schiff Bases/chemistry; Structure-Activity Relationship; Tumor Cells, Cultured; Physicochemical Phenomena
  15. Hoque MM, Omar AR, Hair-Bejo M, Aini I
    J. Biochem. Mol. Biol. Biophys., 2002 Apr;6(2):93-9.
    PMID: 12186763
    Previously we have shown that very virulent infectious bursal disease viruses (vvIBDV) that are SspI, TaqI and StyI positive (92/04, 97/61 and 94/B551) but not SspI and TaqI positive and StyI negative (94/273) cause high mortality, up to 80% in specific-pathogen-free chickens with significant damage of the bursal as well as nonbursal tissues. In this study, we sequenced the VP2 gene (1351 bp) of the 92/04, 94/273 and 94/B551 and compared them with other IBDV strains. All the isolates have the unique amino acid residues at positions 222A, 256I, 294I and 299S found in other vvIBDV strains. The deduced VP2 amino acids encoded by 92/04 is identical to the vvIBDV strains from Israel (IBDVKS), Japan (OKYM) and Europe (UK661), whereas the 94/273 and 94/B551 isolates have one to three amino acid substitutions. The 94/273 has two amino acid substitutions at positions 254 G to S and at 270 A to E that have not been reported before from vvIBDV strains. The 94/B551 also has one amino acid substitution at position 300 E to S, which is uncommon among other vvIBDV strains. However, phylogenetic analysis suggested that the isolates are very close to each other and all of them may have derived from the same origin as vvIBDV strains isolated from China, Japan and Europe. Even though antigenic index analysis of the 94/273 and 94/B551 indicated that the isolates are unique compared to other IBDV strains, their antigenic variation remain to be determined by monoclonal antibody study.
    MeSH terms: Amino Acid Sequence; Animals; Antigens, Viral/genetics; Chickens; DNA, Viral/genetics; Genes, Viral*; Infectious bursal disease virus/genetics*; Infectious bursal disease virus/immunology; Infectious bursal disease virus/isolation & purification; Infectious bursal disease virus/pathogenicity; Molecular Sequence Data; Phylogeny; Virulence/genetics; Viral Structural Proteins/genetics*; Sequence Homology, Amino Acid; Sequence Analysis, DNA; Birnaviridae Infections/virology
  16. Kho CL, Tan WS, Yusoff K
    J. Biochem. Mol. Biol. Biophys., 2002 Apr;6(2):117-21.
    PMID: 12186767
    The phosphoprotein (P) gene of a heat stable Newcastle disease virus (NDV) was cloned, sequenced and expressed in Escherichia coli. SDS-PAGE analysis of the recombinant P protein (395 amino acids) and a C-terminal extension derivative (424 amino acids), gave rise to two distinct protein bands with molecular masses of approximately 53-55 and 56-58 kDa, respectively, which are approximately 26-30% heavier than those calculated from the deduced amino acid sequences. The differences in molecular mass on SDS-PAGE are thought to be attributed to the acidic nature of the P protein (pI=6.27) and also the different degrees of phosphorylation in the prokaryotic cell. Amino acid sequence comparison of the P protein among the published NDV strains showed that they were highly conserved particularly at the putative phosphorylation sites.
    MeSH terms: Amino Acid Sequence; Base Sequence; Cloning, Molecular; DNA, Viral/genetics; Escherichia coli/genetics; Genes, Viral*; Isoelectric Point; Molecular Sequence Data; Molecular Weight; Newcastle disease virus/genetics*; Phosphoproteins/biosynthesis; Phosphoproteins/genetics*; Phosphoproteins/chemistry; Phosphorylation; Plasmids/genetics; Recombinant Proteins/biosynthesis; Recombinant Proteins/genetics; Recombinant Proteins/chemistry; Viral Proteins/biosynthesis; Viral Proteins/genetics*; Viral Proteins/chemistry; Gene Expression; Reverse Transcriptase Polymerase Chain Reaction
  17. Chong SP, Jangi MS, Wan KL
    J. Biochem. Mol. Biol. Biophys., 2002 Apr;6(2):123-8.
    PMID: 12186768
    VCP (Valosin-Containing Protein), a member of the AAA (ATPases Associated to a variety of cellular Activities) family of proteins, possesses a duplicated highly conserved ATPase domain. An expressed sequence tag (EST), representing a clone from the Eimeria tenella merozoite cDNA library, was found to have high similarity to VCP genes from other organisms. A complete sequence derived from the corresponding clone (designated eth060) shows amino acid identity of 42-62% with other members of the VCP subfamily. Sequence analysis identified a putative ATPase domain in the eth060 sequence. This domain was PCR-amplified using gene-specific primers and cloned into a pBAD/Thio-TOPO expression vector. Expression in Escherichia coli demonstrated that the putative ATPase domain, which consists of 414 amino acid residues, produced a fusion protein of approximately 60 kDa in size.
    MeSH terms: Adenosine Triphosphatases/genetics*; Adenosine Triphosphatases/chemistry; Amino Acid Sequence; Animals; Base Sequence; Cloning, Molecular; Escherichia coli; Molecular Sequence Data; Recombinant Fusion Proteins/genetics; Recombinant Fusion Proteins/chemistry; Protozoan Proteins/genetics*; Protozoan Proteins/chemistry; Gene Expression; DNA, Protozoan/genetics; Eimeria tenella/enzymology*; Eimeria tenella/genetics*; Genes, Protozoan; Sequence Homology, Amino Acid; Protein Structure, Tertiary; DNA, Complementary/genetics; Expressed Sequence Tags
  18. Raha AR, Ross E, Yusoff K, Manap MY, Ideris A
    J. Biochem. Mol. Biol. Biophys., 2002 Feb;6(1):7-11.
    PMID: 12186776
    An erythromycin resistance plasmid, pAJ01 was isolated from Loctococcus lactis isolate C5 that was isolated from a healthy two-week-old chicken cecum. A 4 kb plasmid was transformed into plasmidless L. lactis MG1363 before a restriction endonuclease map was constructed. It was then fused with pUC19 to form pAJ02, which can replicate in Escherichia coli XLI-Blue as well as L. lactis MG1363. The plasmid was stably maintained in Lactococcus for more than 100 generations.
    MeSH terms: Animals; Cecum/microbiology*; Chickens; Cloning, Molecular; Erythromycin/pharmacology*; Escherichia coli/genetics; Genetic Vectors*; Plasmids/genetics*; Lactococcus lactis/genetics*; Lactococcus lactis/isolation & purification; Transformation, Bacterial; Restriction Mapping; Drug Resistance, Bacterial
  19. Eshaghi M, Ali AM, Jamal F, Yusoff K
    J. Biochem. Mol. Biol. Biophys., 2002 Feb;6(1):23-8.
    PMID: 12186779
    Streptococcus pyogenes ST4547 is an opacity factor negative strain, which has been recently reported as a new emm type from Malaysia. Nucleotide sequencing of the mga regulon of this strain showed the existence of two emm-like genes. The emm gene located upstream of the scpA gene comprises 1305 nucleotides encoding the putative precursor M protein of 435 amino acids in length with an M(r) of 49 kDa. or a predicted mature protein of 394 amino acids with an M(r) of 44.8 kDa. Another gene mrpST4547 was located upstream of the emm gene and downstream of the mga gene. The sequence of this mrp gene comprises 1167 nucleotides encoding a predicted protein of 388 amino acids in length with an M(r) of 42.2 kDa. or a predicted mature protein of 347 amino acids with an M(r) of 37.9 kDa. The mga regulon of strain ST4547 has a mosaic structure comprising segments, which originated from different OF positive and OF negative strains. The sequences flanking the hyper-variable and C repeats of the emmST4547 gene showed high similarity to corresponding regions in the mga regulon of OF positive strains notably M15, M4, M22 and M50. In contrast, the sequence within the hyper-variable and C repeat regions of the emmST4547 gene revealed high similarity to equivalent regions in the OF negative strains. These data indicates that horizontal transfer of emm-like gene could have occurred between OF positive and OF negative strains resulting in architectural divergence in the mga regulon.
    MeSH terms: Amino Acid Sequence; Antigens, Bacterial*; Bacterial Outer Membrane Proteins/genetics*; Bacterial Proteins*; Base Sequence; Carrier Proteins/genetics*; Codon; DNA, Bacterial/genetics; DNA, Bacterial/isolation & purification; Genes, Bacterial; Membrane Proteins/genetics*; Molecular Sequence Data; Molecular Weight; Phylogeny; Streptococcus pyogenes/genetics*; Open Reading Frames; Regulon*
  20. Nathan S, Li H, Mohamed R, Embi N
    J. Biochem. Mol. Biol. Biophys., 2002 Feb;6(1):45-53.
    PMID: 12186782
    We have used the phagemid pComb3H to construct recombinant phages displaying the single chain variable fragment (ScFv) towards exotoxin of Burkholderia pseudomallei. Variable heavy and light chain fragments were amplified from the hybridoma 6E6A8F3B line, with a wide spectrum of primers specific to mouse antibody genes. Through overlapping extension polymerase chain reaction, the heavy and light chain fragments were linked to form the ScFv which was subsequently cloned into the phage display vector and transformed into ER2537 cells to yield a complexity of 10(8) clones. The transformants were screened by four rounds of biopanning against the exotoxin and resulted in selective enrichment of exotoxin-binding antibodies by 301 fold. The phage pool from the final round of selection displayed antibodies of high-affinity to the exotoxin as demonstrated by ELISA. Several clones were selected randomly from this pool and analysed by restriction enzyme digestion, fingerprinting and sequencing. Restriction analysis confirmed that all clones carried a 700-800 bp insert whose sequences, in general, corresponded to that of mouse IgG. Fingerprinting profiles delineated the antibodies into two families with different CDR sequences.
    MeSH terms: Amino Acid Sequence; Animals; Antibodies, Bacterial/genetics*; Antibodies, Bacterial/immunology; Antibodies, Monoclonal/genetics*; Antibodies, Monoclonal/immunology; Base Sequence; Cell Transformation, Viral; Cloning, Molecular; Enzyme-Linked Immunosorbent Assay; Exotoxins/immunology*; Hybridomas; Immunoglobulin Variable Region/genetics; Immunoglobulin Heavy Chains/genetics; Immunoglobulin Light Chains/genetics; Molecular Sequence Data; Recombinant Fusion Proteins/immunology; Burkholderia pseudomallei/immunology*; Burkholderia pseudomallei/pathogenicity; Peptide Library; Sequence Analysis, Protein; Mice
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