Biomedical subjects

H M Tan

Publications and source records attributed to H M Tan.

At least 37 records · Page 2Linked to original sources

PubMed2000

Transformation of Endomyces fibuliger based on its gene for orotidine-5'-phosphate decarboxylase.

Endomyces fibuliger is a yeast used in the production of Chinese rice wine. It secretes enzymes such as glucoamylase, alpha-amylase and acid protease. Very little is known of the genetics of E. fibuliger. In order to develop a transformation system for this yeast, orotidine-5'-phosphate decarboxylase mutant strains were obtained and characterized. Transformation of the E. fibuliger ura3 mutant F1 with an integrative plasmid that carried the wild-type URA3 gene of E. fibuliger gave complementation of this mutation. The E. fibuliger gene encodes the orotidine-5'-phosphate decarboxylase enzyme consisting of 266 amino acid residues with a 69.4% sequence identity with orotidine-5'-phosphate decarboxylase of Saccharomyces cerevisiae. Our finding that E. fibuliger URA3 complements the ura3 mutation in S. cerevisiae confirms that the URA3 gene of E. fibuliger encodes a protein that exerts a similar function.

Amino Acid Sequence↗

PubMed2000

The genes for benzene catabolism in Pseudomonas putida ML2 are flanked by two copies of the insertion element IS1489, forming a class-I-type catabolic transposon, Tn5542.

Two directly repeated sequences of the IS elements IS1489v1 and IS1489v2 flank the benzene dioxygenase (bedC1C2BA) and the cis-benzene dihydrodiol dehydrogenase (bedD) genes on the catabolic plasmid pHMT112 in Pseudomonas putida ML2, forming a Class-I-type composite transposon, Tn5542. Both IS1489v1 and IS1489v2 contain an identical 1371-bp open reading frame, tnpA, that is preceded by a possible ribosome binding site. The tnpA gene of IS1489v1 is bound by a pair of 40-bp imperfect inverted repeats while that of IS1489v2 is flanked only by the left inverted repeat. The tnpA gene codes for a putative 53-kDa polypeptide of 456 amino acids bearing similarity to transposases encoded on IS elements of P. alcaligenes, P. aeruginosa, P. stutzeri, and Serratia marcescens. The basic nature of the putative TnpA protein with a deduced pI of 8.93 is typical of IS-encoded transposases. Similar to other IS elements, an outward facing promoter was detected at the right end of IS1489v1. Experiments involving the suicide vector, pKNG101, failed to show transposition of Tn5542.

Base Sequence↗

PubMed2000

New therapies for erectile dysfunction.

The quest for improving and maintaining sexual function has been going on since time immemorial. The advent of an effective oral drug, sildenafil, has brought about unprecedented open discussion on male erectile dysfunction, and gas accelerated the pace of development of new therapies for erectile dysfunction. New knowledge in the physiology of sexual function has enabled researchers to target drug treatment at the whole network of the central nervous system and the numerous cascadic enzymatic reactions leading to relaxation of the corporal smooth muscle. One of the brightest potential applications of future molecular technology in the study of erectile dysfuction is in the utilization of gene therapy.

Erectile Dysfunction↗

PubMed2000

Characterization of a predominant immunogenic outer membrane protein of Riemerella anatipestifer.

The ompA gene, encoding the 42-kDa major antigenic outer membrane protein OmpA of Riemerella anatipestifer, the etiololgical agent of septicemia anserum exsudativa, was cloned and expressed in Escherichia coli. Recombinant OmpA displayed a molecular mass similar to that predicted from the nucleotide sequence of the ompA gene but lower than that observed in total cell lysates of R. anatipestifer. The ompA gene showed a conserved C-terminal region comprising the OmpA-like domain and a variable N-terminal region. This structure is similar to those of the analogous outer membrane proteins of several gram-negative bacteria. However, OmpA of R. anatipestifer contains six EF-hand calcium-binding domains and two PEST regions, which distinguish it from other outer membrane proteins. The occurrence of these motifs in OmpA suggests a possible role in virulence for this protein. The ompA gene is present in the R. anatipestifer type strain and in all serotype reference strains. However, it exhibits some minor genetic heterogeneity among different serotypes, which seems not to affect the strong antigenic characteristics of the protein. OmpA is a conserved and strong antigenic determinant of R. anatipestifer and hence is suggested to be a valuable protein for the serodetection of R. anatipestifer infections, independent of their serotype.

Amino Acid Sequence↗

PubMed2000

Penile prosthesis implantation in a transsexual neophallus.

Reconstruction surgery for a female to male transsexual usually involves mastectomy, hysterectomy and creating an aesthetically appealing neophallus. We have successfully inserted an inflatable prosthesis using the AMS CX prosthesis in a 45 year old transsexual, who had a large bulky neophallus constructed from the anterior abdominal subcutaneous fat, about 9 years ago. The single cylinder CX prosthesis was well anchored to the symphysis pubis using a dacron windsock tubing, the activation pump was placed in the dependent pouch of the right labium and the reservior in the usual perivesical space. The patient subsequently had debulking procedure using liposuction to create a more aesthetic and functional phallus. To date, the inflatable neophallus prosthesis is functioning well.

Female↗

PubMed1999

Molecular fingerprinting of Riemerella anatipestifer by repetitive sequence PCR.

Riemerella anatipestifer is a gram-negative rod-shaped bacterium associated with epizootic infections in poultry. A total of 35 R. anatipestifer isolates including the type strain ATCC11845T, reference and field strains for 18 different serotypes were characterized by repetitive sequence based-PCR (rep-PCR) with outwardly-directed primers based on the repetitive extragenic palindromic (REP) consensus sequence. This technique was applied by using either extracted genomic DNA or preparation of whole bacterial cells harvested directly from plate cultures. Rep-PCR discriminated the R. anatipestifer isolates into 19 electrophoretic types. DNA fingerprints obtained from rep-PCR of extracted genomic DNA or from preparations of whole cells yielded comparable patterns. Substantial variation was seen among the rep-PCR fingerprints of different serotypes. Moreover, different polymorphisms of the rep-PCR fingerprints were evident among epidemiologically unrelated isolates of the same serotype. These results suggest the presence of repetitive extragenic palindromic-like elements within the genome of R. anatipestifer that can be used in some isolates to discriminate between different strains belonging to the same serotype. Rep-PCR may serve as a useful molecular tool for subtyping R. anatipestifer isolates for epidemiologic investigations. The whole cell procedure offers the advantage of ease of performance requiring only small quantities of cells.

Animals↗

PubMed1999

Characterization of a novel cis-benzene dihydrodiol dehydrogenase from Pseudomonas putida ML2.

A second and novel cis-benzene dihydrodiol dehydrogenase which is able to dehydrogenate a range of cis-dihydrodiols and other vicinal alcohols has been purified from Pseudomonas putida ML2. The enzyme is a tetramer of a polypeptide of 39 kDa in molecular mass and has a pH optimum of 9.0. Despite having a primary structure that has significant similarity to glycerol dehydrogenases, the kcat/Km value of the enzyme for cis-benzene dihydrodiol is 4300-fold higher compared to glycerol. The apparent Km values of the enzyme for cis-benzene dihydrodiol and glycerol are 0.01 mM and 46 mM, respectively, and 0.22 mM for NAD+.

Amino Acid Sequence↗

PubMed1999

Bacterial catabolic transposons.

The introduction of foreign organic hydrocarbons into the environment in recent years, as in the widespread use of antibiotics, has resulted in the evolution of novel adaptive mechanisms by bacteria for the biodegradation of the organic pollutants. Plasmids have been implicated in the catabolism of many of these complex xenobiotics. The catabolic genes are prone to undergo genetic rearrangement and this is due to their presence on transposons or their association with transposable elements. Most of the catabolic transposons have structural features of the class I (composite) elements. These include transposons for chlorobenzoate (Tn5271), chlorobenzene (Tn5280), the newly discovered benzene catabolic transposon (Tn5542), and transposons encoding halogenated alkanoates and nylon-oligomer-degradative genes. Transposons for the catabolism of toluene (Tn4651, Tn4653, Tn4656) and naphthalene (Tn4655) belong to class II (Tn3 family) elements. Many catabolic genes have been associated with insertion sequences, which suggests that these gene clusters could be rapidly disseminated among the bacterial populations. This greatly expands the substrate range of the microorganisms in the environment and aids the evolution of new and novel degradative pathways. This enhanced metabolic versatility can be exploited for and is believed to play a major part in the bioremediation of polluted environments.

Bacteria↗

PubMed1999

Mobile catabolic genes in bacteria.

The recent findings of various mobile catabolic genes have provided some insight into the evolution of microbial degradation systems for xenobiotic compounds. The catabolic genes undergo marked genetic rearrangements due to their presence on transposons or association with mobile genetic elements. Bacterial catabolic transposons fall into three defined structural classes. Class I elements include catabolic genes flanked by two copies of insertion sequences. Class II elements carry short terminal inverted repeats and transpose by the replicative mode in which transposase and resolvase are involved. Conjugative catabolic transposons represent the third class of mobile genetic elements. They carry all the genes required for excision, conjugal transfer to a new host, and integration. This review focuses on the structures, functions and roles of the recently characterized catabolic transposons in bacteria. Also described are the mobile catabolic elements that share structural similarity with the pathogenicity and symbiosis islands.

Journal Article↗

PubMed1997

Phylogenetic position of Riemerella anatipestifer based on 16S rRNA gene sequences.

Riemerella anatipestifer, the causative agent of septicemia anserum exsudativa (also called new duckling disease), belongs to the family Flavobacteriaceae of gram-negative bacteria. We determined the DNA sequences of the rrs genes encoding the 16S rRNAs of four R. anatipestifer strains by directly sequencing PCR-amplified rrs genes. A sequence similarity analysis confirmed the phylogenetic position of R. anatipestifer in the family Flavobacteriaceae in rRNA superfamily V and allowed fine mapping of R. anatipestifer on a separate rRNA branch comprising the most closely related species, Bergeyella zoohelcum, as well as Chryseobacterium balustinum, Chryseobacterium indologenes, and Chryseobacterium gleum. The sequences of the rrs genes of the four R. anatipestifer strains varied between 0.5 and 1.0%, but all of the strains occupied the same position on the phylogenetic tree. In general, differences in rrs genes were observed among R. anatipestifer strains, even within a given serotype, as shown by restriction fragment length polymorphism of PCR-amplified rrs genes.

Animals↗

PubMed1997

A comparison of antiemetic efficacy of droperidol alone and in combination with metoclopramide in day surgery anaesthesia.

We have studied the antiemetic efficacy of droperidol alone, and in combination with metoclopramide in first trimester termination of pregnancy in day surgery. The aim was to determine whether the addition of metoclopramide could further reduce the incidence of postoperative nausea and vomiting (PONV) but avoid excessive sedation. Group I (control, n = 40) received i.v. droperidol 0.625 mg at induction. Group II (study, n = 40) received i.v. droperidol 0.625 mg and i.v. metoclopramide 10 mg at induction. The incidence of nausea at 1 and 2 hours postoperatively was 23% and 10% in group I, and 5% and nil in group II respectively. The difference in the incidence of nausea was significant at p < 0.05 at one hour but not at two hours postoperatively. No patients vomited. There was no difference in the sedation and pain score between them. We did not observe any significant side effects attributable to either drug. All patients were discharged home within 3 hours. We conclude that in the prevention of PONV, the combination of metoclopramide and droperidol is superior to the use of droperidol alone at one hour but not at two hours postoperatively.

Adult↗

PubMed1997

Sedation for the conduct of lumbar epidural anaesthesia: a study using subanaesthetic dose of ketamine in combination with midazolam.

We gave sedation for the conduct of lumbar epidural analgesia using intravenous ketamine 0.3 mg.kg-1 with intravenous midazolam 2 mg. Forty adult Chinese females undergoing major gynaecological laparotomies had epidural catheter inserted before general anaesthesia, 20 of whom were given ketamine and midazolam (study group) and the other 20 acted as control. During the conduct of the epidural, the pain and anxiety scores in the study group were significantly less than the control group (P < 0.05). Patients were significantly more sedated in the study group (P < 0.05). All the patients in the study group were satisfied and would consent to future epidural versus 75% in the control group (significant at P < 0.05). Ninety per cent of patients in the study group had amnesia but none in the control group. Pain experienced during the epidural was the reason for refusal of future epidural. We did not observe any emergence phenomenon or cardiovascular stimulation. There was a statistically significant decrease in the pulse oximetry oxygen saturation (SpO2) in the study group but none required oxygen supplementation. We concluded that pain caused by the conduct of epidural did decrease the patient's acceptance rate to future epidural, and the combined use of intravenous ketamine 0.3 mg.kg-1 and midazolam 2 mg provided adequate sedation, analgesia, anxiolysis and amnesia to significantly increase the acceptance rate without any significant side effects.

Adult↗

PubMed1996

Characterization and expression of the plasmid-borne bedD gene from Pseudomonas putida ML2, which codes for a NAD+-dependent cis-benzene dihydrodiol dehydrogenase.

The catabolic plasmid pHMT112 in Pseudomonas putida ML2 contains the bed gene cluster encoding benzene dioxygenase (bedC1C2BA) and a NAD+-dependent dehydrogenase (bedD) required to convert benzene into catechol. Analysis of the nucleotide sequence upstream of the benzene dioxygenase gene cluster (bedC1C2BA) revealed a 1,098-bp open reading frame (bedD) flanked by two 42-bp direct repeats, each containing a 14-bp sequence identical to the inverted repeat of IS26. In vitro translation analysis showed bedD to code for a polypeptide of ca. 39 kDa. Both the nucleotide and the deduced amino acid sequences show significant identity to sequences of glycerol dehydrogenases from Escherichia coli, Citrobacter freundii, and Bacillus stearothermophilus. A bedD mutant of P. putida ML2 in which the gene was disrupted by a kanamycin resistance cassette was unable to utilize benzene for growth. The bedD gene product was found to complement the todD mutation in P. putida 39/D, the latter defective in the analogous cis-toluene dihydrodiol dehydrogenase. The dehydrogenase encoded by bedD) was overexpressed in Escherichia coli and purified. It was found to utilize NAD+ as an electron acceptor and exhibited higher substrate specificity for cis-benzene dihydrodiol and 1,2-propanediol compared with glycerol. Such a medium-chain dehydrogenase is the first to be reported for a Pseudomonas species, and its association with an aromatic ring-hydroxylating dioxygenase is unique among bacterial species capable of metabolizing aromatic hydrocarbons.

Amino Acid Sequence↗

PubMed1995

A single base-pair change (ATG-->ATC) nullifies the activity of cytosolic fumarase in Saccharomyces cerevisiae.

A respiratory defective pet mutant (W303 delta FUM1) of Saccharomyces cerevisiae deficient in fumarase was transformed with the plasmid construct pG5/ST7. This plasmid contains the entire FUM1 gene with only a single base pair change (ATG-->ATC) confined to the putative second inframe translation initiation codon. While transformation of the fumarase deficient mutant with pG5/ST7 resulted in an elevation of fumarase activity in the mitochondria of the transformed strain, fumarase activity in the cytosol remained negligible his result indicated that the cytosolic fumarase isoenzyme is synthesized exclusively from the second translation initiation codon of FUM1. Results of RACE-PCR of the 5' ends of FUM1 transcripts confirmed that the FUM1 gene synthesizes two distinct species of transcripts. These data provide strong evidence for the synthesis and targeting of two fumarase isomers.

Base Composition↗

PubMed1995

Multimodal approach in the management of 1163 ureteric stone cases.

One thousand one hundred and sixty three patients (male-852, female-311) with ureteric calculi requiring intervention were treated between April 1988 to July 1992. Four hundred and eleven cases were treated by ESWL Monotherapy, 414 by stone manipulation plus ESWL, 301 by retrograde ureteroscopic lithotripsy, 36 by percutaneous antegrade ureteroscopic lithotripsy and 1 case by open ureterolithotomy. There were 25 failures of the initial procedures. Only three cases that failed primary procedures required open surgery. Other complications include minor ureteric mucosal perforation (3%), infection (3%), transient moderate to gross haematuria (20%), loin ache (26.4%), irritative urination (34.4%) and low grade fever (30.1%). Current modalities used in the treatment of ureteric calculi produce good results and there is generally no primary role for any open surgery.

Adolescent↗

PubMed1994

YKE2, a yeast nuclear gene encoding a protein showing homology to mouse KE2 and containing a putative leucine-zipper motif.

The nucleotide sequence of YKE2, a yeast nuclear gene, has been determined. The deduced YKE2 protein has 114 amino acids (12 kDa), shows significant homology with the murine KE2 and contains a putative Leu-zipper motif characteristic of a group of DNA-binding proteins [Landschulz et al., Science 240 (1988) 1759-1764]. Strains in which YKE2 has been disrupted show normal cell growth in glucose and galactose media over the temperature range of 16 to 40 degrees C. Disrupted strains also display normal mating and sporulation abilities. Northern analysis revealed that the transcription of YKE2 is unresponsive to catabolite repression by glucose.

Amino Acid Sequence↗

PubMed1994

The Saccharomyces cerevisiae homologue of ribosomal protein S26.

The nucleotide sequence of RPS26, the gene encoding a homologue of ribosomal protein small subunit S26 in Saccharomyces cerevisiae, was determined. The deduced amino-acid sequence showed significant identity with its counterparts from Neurospora crassa, human, rat and Arabidopsis thaliana. Disruption of RPS26 resulted in the formation of micro-colonies, suggesting that it is important for the normal cell growth of S. cerevisiae.

Amino Acid Sequence↗

PubMed1994

Substitution of the ISP alpha subunit of biphenyl dioxygenase from Pseudomonas results in a modification of the enzyme activity.

We have constructed a hybrid multicomponent dioxygenase gene cluster in which the bphA1 gene, coding for ISP alpha subunit of biphenyl dioxygenase from Pseudomonas pseudoalcaligenes KF707, has been replaced by the bedC1 gene encoding the corresponding subunit of benzene dioxygenase from P. putida ML2. Escherichia coli cells containing the chimeric dioxygenase acquired the novel capability of producing indigo from indole. Furthermore, when compared to biphenyl dioxygenase, the hybrid dioxygenase enzyme was half as active towards benzene but exhibited only 4% activity when biphenyl was the substrate. The results implicate the ISP alpha subunit to be involved in substrate specificity and activity of the dioxygenase enzyme.

Escherichia coli↗