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Biomedical subjects

E W Chan

Publications and source records attributed to E W Chan.

16 recordsLinked to original sources

What have we learnt from community-acquired infections in Hong Kong?

This study was initiated throughout Hong Kong, to reveal the characteristics of community-acquired infections. All specimens collected by general practitioners from infected patients were followed prospectively, and those that were culture-positive were analysed. Four thousand seven hundred and forty-one specimens were collected from 3977 patients by 89 doctors from July 2000 to October 2001. The most common specimens were throat swabs (33%), urine (26%) and sputa (16%). The average culture-positive rate was 28%. The most common organisms were Escherichia coli (18%), beta-haemolytic streptococci (15%) and Staphylococcus aureus (12%). Fluoroquinolone resistance was relatively high (up to 35%) in organisms commonly causing urinary tract infection (E. coli, Proteus and Morganella). Although none of the pneumococci was resistant to penicillin 1 mg/L, the proportion with intermediate resistance (0.1-1 mg/L) was alarming (81%). There were three strains of methicillin-resistant S. aureus. A decrease in ampicillin resistance but a high prevalence of macrolide resistance were noted in Haemophilus influenzae. All Neisseria gonorrhoeae isolates were resistant to penicillin, up to 79% to the fluoroquinolones, 15% to spectinomycin, but all were susceptible to ceftriaxone. Respiratory pathogens (Streptococcus pneumoniae, beta-haemolytic streptococci and H. influenzae) were relatively susceptible to the newer fluoroquinolones (0-2%, 0.5-6% and 2% resistant, respectively) or third-generation cephalosporins (0-2% resistant). The distribution of organisms and their antibiotic resistance varied over time. Thus frequent surveillance is needed to provide information on the drugs of choice for different infections.

Adolescent↗

Mutations in topoisomerase genes of fluoroquinolone-resistant salmonellae in Hong Kong.

A total of 88 salmonella isolates (72 clinical isolates for which the ciprofloxacin MIC was >0.06 microg/ml, 15 isolates for which the ciprofloxacin MIC was < or =0.06 microg/ml, and Salmonella enterica serotype Typhimurium ATCC 13311) were studied for the presence of genetic alterations in four quinolone resistance genes, gyrA, gyrB, parC, and parE, by multiplex PCR amplimer conformation analysis. The genetic alterations were confirmed by direct nucleotide sequencing. A considerable number of strains had a mutation in parC, the first to be reported in salmonellae. Seven of the isolates sensitive to 0.06 micro g of ciprofloxacin per ml had a novel mutation at codon 57 of parC (Tyr57-->Ser) which was also found in 29 isolates for which ciprofloxacin MICs were >0.06 micro g/ml. Thirty-two isolates had a single gyrA mutation (Ser83-->Phe, Ser83-->Tyr, Asp87-->Asn, Asp87-->Tyr, or Asp87-->Gly), 34 had both a gyrA mutation and a parC mutation (29 isolates with a parC mutation of Tyr57-->Ser and 5 isolates with a parC mutation of Ser80-->Arg). Six isolates which were isolated recently (from 1998 to 2001) were resistant to 4 micro g of ciprofloxacin per ml. Two of these isolates had double gyrA mutations (Ser83-->Phe and Asp87-->Asn) and a parC mutation (Ser80-->Arg) (MICs, 8 to 32 microg/ml), and four of these isolates had double gyrA mutations (Ser83-->Phe and Asp87-->Gly), one parC mutation (Ser80-->Arg), and one parE mutation (Ser458-->Pro) (MICs, 16 to 64 micro g/ml). All six of these isolates and those with a Ser80-->Arg parC mutation were S. enterica serotype Typhimurium. One S. enterica serotype Typhi isolate harbored a single gyrA mutation (Ser83-->Phe), and an S. enterica serotype Paratyphi A isolate harbored a gyrA mutation (Ser83-->Tyr) and a parC mutation (Tyr57-->Ser); both of these isolates had decreased susceptibilities to the fluoroquinolones. The MICs of ciprofloxacin, levofloxacin, and sparfloxacin were in general the lowest of those of the six fluoroquinolones tested. Isolates with a single gyrA mutation were less resistant to fluoroquinolones than those with an additional parC mutation (Tyr57-->Ser or Ser80-->Arg), while those with double gyrA mutations were more resistant.

Anti-Infective Agents↗

Triptolide induced cytotoxic effects on human promyelocytic leukemia, T cell lymphoma and human hepatocellular carcinoma cell lines.

Triptolide, a traditional Chinese medicine, has been reported to be effective in the treatment of auto-immune diseases, and it can also induce anti-neoplastic activity on several human tumor cell lines. This study investigates the cytotoxic function and the functional mechanism of triptolide on tumor cells. Promyelocytic leukemia, (HL-60), T cell lymphoma (Jurkat), and human hepatocelluar carcinoma (SMMC-7721) cells were subjected to triptolide treatment, and cell growth inhibition was examined by XTT cell viability assay. Cell death mechanism (apoptosis) was confirmed through DNA fragmentation and DAPI staining. Triptolide inhibited 50% of cell growth (IC(50)) on HL-60 cells at 7.5 nM, Jurkat cells at 27.5 nM and SMMC cells at 32 nM. Characteristic apoptotic features including internucleosomal DNA fragmentation and chromatin condensation were observed in triptolide treated cells. Data from the study indicates that triptolide could induce apoptosis in human tumor cell lines and it may be applicable as a potential chemotherapeutic agent for cancer treatment.

Antineoplastic Agents, Alkylating↗

Molecular epidemiological analysis of Salmonella enterica serotype Derby infections in Hong Kong.

OBJECTIVES: We aimed to study the antimicrobial susceptibilities and molecular epidemiology of Salmonella enterica serotype Derby, a unique and common salmonella serotype in Hong Kong. METHODS: Salmonella Derby strains isolated from stools of patients in a large general hospital in Hong Kong from 1989 to 1994 and from food samples isolated in the Public Health Laboratory were randomly selected and investigated for the antimicrobial susceptibilities by determining the minimal inhibitory concentrations of 19 antimicrobial agents and their relatedness using plasmid analysis, ribotyping, pulsed-field gel electrophoresis (PFGE) and total DNA fingerprinting. RESULTS: About 50% of the 127 isolates studied were susceptible to all the 19 antibiotics tested, although resistance to tetracycline (49%) and sulfamethoxazole (38%) was high. Only 12% did not harbour any detectable plasmids, while the rest contained plasmids in 51 profiles. There were two predominant clones, one comprising of 35% of isolates that could not be pulsotyped because discrete bands were not discernible after PFGE and another comprising 34% of isolates that could be pulsotyped. The remaining 31% belonged to a variety of types. CONCLUSIONS: Approximately 70% of S. Derby belonging to two clones were endemic in the community, while the remaining isolates belonged to a variety of types which were probably a result of sporadic infection. The sources of human infections were foods, since most isolates from foods also belonged to the two endemic clones. Typing of S. Derby isolates from other sources such as animals or the environment would help elucidate how foods were contaminated. PFGE might not be universally applicable to all salmonella strains.

Adolescent↗

Plasmids encoding foot-and-mouth disease virus VP1 epitopes elicited immune responses in mice and swine and protected swine against viral infection.

VP1 is a capsid protein of foot-and-mouth disease virus (FMDV) and contains epitopes of the virus. Plasmids encoding two VP1 epitopes (amino acid residues 141-160 and 200-213) and a host-self immunoglobulin molecule were constructed to produce a new type of FMD DNA vaccine. Two plasmids, namely, pCEIM and pCEIS, containing mouse immunoglobulin (IgG) or swine IgG were subjected to immunogenicity testing in mice and swine, respectively. In mice administrated pCEIM in the abdomen using a genegun, both FMDV-specific T-cell proliferation and neutralizing antibodies were detected. In swine immunized with pCEIS at the back of the ear, immune responses were achieved after the second administration. Swine showed a T-cell proliferative response with a stimulation index (SI) of up to 8.1 and a neutralizing antibody response that was able to protect suckling mice from 10(2) LD(50) (lethal dose 50) FMDV challenge. To compare the immunogenicity of the DNA-based vaccine candidate, versus the protein-based vaccine candidates, a second group of swine was immunized with the protein F1-scIgG, which was encoded by the plasmid pCEIS. Injection with F1-scIgG elicited a T-cell proliferative response of SI < 1.7 and a neutralizing antibody response that protected suckling mice from up to 10(5) LD(50) FMDV challenge. In the challenge test, three of three swine immunized with pCEIS were fully protected from FMDV challenge.

Animals↗

An immunoglobulin G based chimeric protein induced foot-and-mouth disease specific immune response in swine.

Epitopes containing the residues 141aa-160aa and 200aa-213aa from foot-and-mouth disease (FMD) serotype O1K HK type FMDV VP1 were joined to a swine immunoglobulin G single heavy chain constant region (scIgG), creating a novel chimeric protein, named F1-scIgG. In this study, inoculation with F1-scIgG induced both FMD virus-neutralizing antibody response and T cell response in swine. Antisera from these F1-scIgG-inoculated swine protected suckling mice against 1000 lethal dose 50 (1000LD(50)) FMD challenge. F1-scIgG-inoculated swine were also fully protected against 50LD(50) FMD virus challenge. The present study demonstrates the clear potential for viral epitopes linked with self-Ig in novel FMD vaccine design.

Animals↗

Stability of cefazolin sodium eye drops.

OBJECTIVE: Assessing the stability of cefazolin sodium in preservative-free and preservative-containing eye drops. METHOD: Extemporaneous formulations of eye drops were prepared from a commercially-available parenteral product of cefazolin sodium: eye drops 'A' contained 50mg/ml of cefazolin sodium in 0.45% w/v sodium chloride solution, and eye drops 'B' contained 50 mg/ml, 0.005% w/v thiomersal and 1% w/v glycerol in water-for-injection. Cefazolin sodium concentrations in these eye drops were monitored by a stability-indicating HPLC assay method. Measurements of pH and osmolality, as well as tests for microbial contamination, were conducted. RESULTS: The eye drops stored at 4 degrees C were stable for 42 days with minimal changes in pH and osmolality, but eye drops stored at room temperature were only stable for a few days with greater increments in pH and osmolality. None of the samples cultured had bacterial or fungal growth after 7 days of incubation. CONCLUSION: Extemporaneously prepared formulations of cefazolin are unstable at room temperature and should be stored in a refrigerator.

Bacteriological Techniques↗

Stability of phenoxybenzamine hydrochloride in various vehicles.

The stability of phenoxybenzamine hydrochloride in various oral liquids was studied. Phenoxybenzamine hydrochloride powder or capsules were combined with various vehicles to prepare 10-mL formulations with a drug concentration of 2 mg/mL and a 20-mL stock solution containing 10 mg/mL. All formulations were prepared in triplicate and stored at 4 degrees C. A 1.0-mL sample of each of the 2-mg/mL formulations was withdrawn on days 0, 1, 2, 3, and 4, and samples of selected formulations were taken on days 7 and 10. Samples of the stock solution were withdrawn on days 0, 2, 4, 7, 10, 14, and 30. All samples were analyzed by high-performance liquid chromatography. Phenoxybenzamine hydrochloride 2 mg/mL was stable for a longer time in 1% propylene glycol, 0.15% citric acid, and water than in a similar vehicle in which syrup was used in place of water. The concentrations of the drug in both the 10-mg/mL stock solution and stock solutions diluted to 2 mg/mL were > 90% of the initial drug concentration for 30 days. Phenoxybenzamine hydrochloride 2 mg/mL in 1% propylene glycol and 0.15% citric acid in distilled water was stable for 7 days at 4 degrees C. A stock solution of phenoxybenzamine hydrochloride 10 mg/mL in propylene glycol was stable for 30 days at 4 degrees C and, after dilution with 66.7% sucrose in distilled water to a concentration of 2 mg/mL, for up to one hour at 4 degrees C.

Chromatography, High Pressure Liquid↗

Antiviral properties of polyinosinic acids containing thio and methyl substitutions.

Polyinosinic acids containing methyl and sulphur substitutions are potent inhibitors of reverse transcriptase. Substitution of sulphur for oxygen at the 6 position produces significant effects on the properties of polyinosinic acid: the kinetics of inhibition change from competitive to mixed-type and the inhibition constant falls by three orders of magnitude. In contrast, 1-methyl substitution produces no such effects. Poly(1-methyl-6-thioinosinic acid) or poly(m1s6I) inhibits irreversibly, inhibiting all ten reverse transcriptases tested under a variety of assay conditions. In cell culture test systems, poly(m1s6I) is capable of blocking both infection by non-transforming viruses and transformation by a sarcoma virus. The presence of poly(m1s6I) in a preinfected culture results in the production of non-infectious virus particles lacking reverse transcriptase activity.

Avian Myeloblastosis Virus↗

Effects of polyethylene glycol on reverse transcriptase and other polymerase activities.

Polyethylene glycol enhances reverse transcription, augmenting both the rate and duration of polymerization. The effective mean molecular weight of polyethylene glycol is 6000 and the optimal concentration is 12% (w/w). Polyethylene glycol is effective on the reverse transcriptase reaction of all ten type B, C, and D viruses tested under a variety of exogenous, endogenous, and reconstitution assay systems, including the highly efficient conditions involving calf thymus DNA oligonucleotide primers. By three methods of synthesis, polyethylene glycol increased the yields of complementary [3H]DNA by a factor of 1.8--6.5. Polyethylene glycol does not alter the divalent cation requirements of the specificities of the enzyme. Complementary [3H]DNAs made in the presence of polyethylene glycol are indistinguishable in terms of size and sequence complementarity from those made in the absence of the polymer. The stimulatory effect was partly due to the ability of polyethylene glycol to stabilize reverse transcriptase. Preliminary tests indicate that polyethylene glycol also stimulates other nucleotide polymerases, such as the DNA-dependent DNA and RNA polymerases of Escherichia coli and the terminal transferase of calf thymus.

Escherichia coli↗

Microtissue culture plaque assay for Herpesvirus saimiri.

A microtissue culture method for the plaque assay of Herpesvirus saimiri has been developed. Virus titrations carried out in Microtest II tissue culture plates (Falcon) yielded reproducible results that agreed well with those obtained by employing macrocultures. The described method is quantitative, reproducible, economical, and suitable for routine assay of large numbers of virus samples.

Adsorption↗