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Biomedical subjects

S W Chan

Publications and source records attributed to S W Chan.

At least 55 records · Page 3Linked to original sources
PubMed1996

Determination of the L-ascorbic acid requirements in Wistar osteogenic disorder Shionogi rats for prolonged carcinogenesis experiments.

Wistar Shionogi rats of the (od/od) substrain with the osteogenic disorder are unable to synthesize L-ascorbic acid (L-AA) and appear to be an appropriate animal model for studying the effect of L-AA in carcinogenesis. To determine the minimal L-AA requirements of these animals for prolonged survival in a satisfactory physical condition during experimentation, four concentrations of L-AA (0.33 g/l, 0.67 g/l, 1.67 g/l and 3.33 g/l) were administered via drinking water to four groups of animals (n = 2). Their water intake per cage was recorded three times weekly and the plasma L-AA levels were determined at the start, after 2, 4, 8 and 12 weeks and at the termination of the experiment. To simulate the procedures to be undertaken in oral mucosal carcinogenesis experiments, the animals were gently restrained and a designated amount of sterile NaCl was applied to the palatal mucosa three times a week for 26 weeks. The L-AA supplement group with the lowest concentration (0.33 g/l L-AA) achieved mean plasma levels of 7 +/- 1.38 microM, approximately one-eighth that of the normal level (mean plasma L-AA level in outbred Wistar rats was found to be 58 +/- 3 microM) whilst those in the higher supplement group (3.33 g/l L-AA) achieved a mean of 18 +/- 1.25 microM. All of the animals employed in the present study survived for 26 weeks and showed no clinical signs of L-AA deficiency during this period.

Animals↗
PubMed1994

Comparison of Giemsa and fluorescent monoclonal antibody staining of inoculated cell cultures for diagnosis of Chlamydia trachomatis.

Three methods of isolation and identification of Chlamydia trachomatis inclusions in cell monolayers were compared: Giemsa staining followed by darkfield microscopy of cycloheximide treated Buffalo green monkey kidney monolayers (BGM), fluorescent monoclonal antibody staining of cycloheximide treated BGM and of HeLa 229 monolayers. In an unselected group of 895 patients with suspected chlamydial infections including contacts of symptomatic patients, significantly more positive chlamydia isolates were detected with immunofluorescent monoclonal antibody staining of BGM and HeLa 229 than with Giemsa staining followed by darkfield microscopy on BGM. However, there was no disparity between the 2 staining methods with specimens from 156 patients with untreated non-gonococcal urethritis. In all chlamydia positive specimens, significantly more inclusions were seen after BGM-immunofluorescence than after HeLa-immunofluorescence or BGM-Giemsa. The extra cost associated with immunofluorescence staining was justified by the time saved in scanning.

Animals↗
PubMed1993

Detection of Chlamydia trachomatis in urine using enzyme immunoassay and DNA amplification.

The suitability of urine specimens for the detection of Chlamydia trachomatis infections in men was assessed. Urethral swabs from 301 patients were cultured for C. trachomatis, and the results were compared with results obtained from Chlamydiazyme. The results of 298 specimens were also compared with results obtained from PCR analysis of first-void urine specimens. The sensitivity of confirmed Chlamydiazyme analysis was 93% and the specificity was greater than 99% compared with culture. The sensitivity of the PCR method was 100% compared with culture. Chlamydia trachomatis was detected by PCR in an additional three specimens from which C. trachomatis could not be cultured. Urine appears to be an appropriate specimen for the detection of C. trachomatis antigens and nucleic acids.

Adolescent↗
PubMed1993

Detection of three types of hepatitis C virus in blood donors: investigation of type-specific differences in serologic reactivity and rate of alanine aminotransferase abnormalities.

The serologic reactivity and epidemiology associated with different hepatitis C virus (HCV) variants were investigated in a cohort of 113 anti-HCV-positive donors. In Scotland, HCV type 1 accounted for one-half of all infections; 40 percent of subjects were infected with HCV type 3, and the remainder were infected with type 2. Reactivity with the NS-4-encoded antigens in the first-generation anti-c100 assay was absent in 68 percent of donors infected with types 2 and 3, as compared with 10 percent for those infected with type 1. Even when combined with surrogate marker testing, first-generation tests would have failed to detect 12 percent of HCV-infected blood donors. The age distribution, incidence of past infection with hepatitis B virus, and reported risk factors were similar in donors infected with types 1 and 3 (mean ages were 31.9 and 29.9; 18 and 17.5% were positive for antibody to hepatitis B core antigen; and 47 and 48% had past intravenous drug abuse). However, the distributions of alanine aminotransferase levels were significantly different in those infected with type 3 (abnormally raised in 83%) and those infected with type 1 (55% abnormal alanine aminotransferase; p < 0.05) or type 2 (60%; p < 0.01) and those who were nonviremic (8%; p < 0.0001). These data suggest that HCV type 1 is the most common HCV infection in blood donors and that infection with HCV type 3 may be associated with more severe liver disease, because of more recent infection or because of a greater inherent pathogenicity of type 3 variants.

Alanine Transaminase↗
PubMed1993

Classification of hepatitis C virus into six major genotypes and a series of subtypes by phylogenetic analysis of the NS-5 region.

Hepatitis C virus (HCV) showed substantial nucleotide sequence diversity distributed throughout the viral genome, with many variants showing only 68 to 79% overall sequence similarity to one another. Phylogenetic analysis of nucleotide sequences derived from part of the gene encoding a non-structural protein (NS-5) has provided evidence for six major genotypes of HCV amongst a worldwide collection of 76 samples from HCV-infected blood donors and patients with chronic hepatitis. Many of these HCV types comprised a number of more closely related subtypes, leading to a current total of 11 genetically distinct viral populations. Phylogenetic analysis of other regions of the viral genome produced relationships between published sequences equivalent to those found in NS-5, apart from the more highly conserved 5' non-coding region in which only the six major HCV types, but not subtypes, could be differentiated. A new nomenclature for HCV variants is proposed in this communication that reflects the two-tiered nature of sequence differences between different viral isolates. The scheme classifies all known HCV variants to date, and describes criteria that would enable new variants to be assigned within the classification as they are discovered.

Base Sequence↗
PubMed1993

Sequence variability in the 5' non-coding region of hepatitis C virus: identification of a new virus type and restrictions on sequence diversity.

We have analysed the pattern of nucleotide sequence variability in the 5' non-coding region (5' NCR) of geographically dispersed variants of hepatitis C virus (HCV). Phylogenetic analysis of sequences in this region indicated the existence of a new virus type, provisionally termed type 4, the identity of which was confirmed by further analysis of the more variable part of the HCV core protein coding region. The geographical distribution of HCV type 4 was distinct from that of other HCV types, it being particularly widespread in Africa and absent or rare in Europe and the Far East. Much of the variability in the 5' NCR appears to be constrained by a requirement for specific secondary structures in the viral RNA. In one of the most variable regions of the 5' NCR (positions -169 to -114), most of the nucleotide changes that are characteristic of different HCV types were covariant, with complementary substitutions at other positions. According to the proposed secondary structure of the 5' NCR, such changes preserved base pairing within a stem-loop structure, whereas the nucleotide insertions found in a proportion of 5' NCR sequences, including those of type 4, localized exclusively to the non-base-paired terminal loop. The specific nucleotide substitutions in the 5' NCR that differentiate each of the four HCV types can be detected by restriction enzyme cleavage, providing a rapid and reliable method for virus typing.

Amino Acid Sequence↗
PubMed1993

Mapping of serotype-specific, immunodominant epitopes in the NS-4 region of hepatitis C virus (HCV): use of type-specific peptides to serologically differentiate infections with HCV types 1, 2, and 3.

The effect of sequence variability between different types of hepatitis C virus (HCV) on the antigenicity of the NS-4 protein was investigated by epitope mapping and by enzyme-linked immunosorbent assay with branched oligopeptides. Epitope mapping of the region between amino acid residues 1679 and 1768 in the HCV polyprotein revealed two major antigenic regions (1961 to 1708 and 1710 to 1728) that were recognized by antibody elicited upon natural infection of HCV. The antigenic regions were highly variable between variants of HCV, with only 50 to 60% amino acid sequence similarity between types 1, 2, and 3. Although limited serological cross-reactivity between HCV types was detected between peptides, particularly in the first antigenic region of NS-4, type-specific reactivity formed the principal component of the natural humoral immune response to NS-4. Type-specific antibody to particular HCV types was detected in 89% of the samples from anti-HCV-positive blood donors and correlated almost exactly with genotypic analysis of HCV sequences amplified from the samples by polymerase chain reaction. Whereas almost all blood donors appeared to be infected with a single virus type (97%), a higher proportion of samples (40%) from hemophiliacs infected from transfusion of non-heat-inactivated clotting factor contained antibody to two or even all three HCV types, providing evidence that long-term exposure may lead to multiple infection with different variants of HCV.

Amino Acid Sequence↗
PubMed1993

Evaluation of Pharmacia Chlamydia EIA for diagnosis of genital infections caused by Chlamydia trachomatis.

A new enzyme-immunoassay test (EIA), Pharmacia Chlamydia EIA, was evaluated in comparison with isolation in cell culture for the diagnosis of genital infections caused by Chlamydia trachomatis. Endocervical and endourethral specimens were collected from 913 patients (875 female and 38 male patients respectively). Sensitivity, specificity, positive predictive and negative predictive values for this test were 89.4%, 99.2%, 85.8% and 99.4% respectively. Where discrepancies between the 2 techniques occurred, the remainder of the EIA buffer was stained for chlamydial antigen by direct immunofluorescence using the Syva Microtrak Chlamydia trachomatis Direct Specimen Test. Laboratory strains of various micro-organisms that may be present in genital specimens did not cross react with the EIA test except for Staphylococcus aureus and Group B Streptococcus at very high concentrations. The Pharmacia Chlamydia EIA is suitable for a laboratory which handles a large number of chlamydial specimens daily. However, when interpreting EIA results, the prevalence of chlamydial infection in the patient population should be established. At present, EIA tests are unsuitable as the only test for chlamydial diagnosis in low risk populations.

Bacteriological Techniques↗
PubMed1992

Pyrimethamine resistant mutations in Plasmodium falciparum.

Three mutations in Plasmodium falciparum yielding increased resistance to pyrimethamine were obtained following treatment with chemical mutagens and selection in presence of pyrimethamine. From parasite clone TM4/8.2 a mutant, TM4/8.2/4.1, was produced which raised pyrimethamine resistance about 500 times and was found to involve an amino acid change in the DHFR-TS enzyme molecule from Ser108 to Asn108. A clone of another isolate, T9/94, yielded a mutant, T9/94/300.300, raising pyrimethamine resistance about 10 times and involving an amino acid change from Ile164 to Met164. However, another mutant from T9/94, T9/94/M1-1(b3), although it raised the pyrimethamine resistance 100 times, did not involve any changes in the coding sequence of the DHFR-TS gene, but resulted in the production of about twice as much DHFR-TS enzyme as the original clone T9/94. No amplification of the DHFR-TS gene was detected. It is concluded that changes in pyrimethamine resistance of malaria parasites may arise in at least 2 ways: (1) by structural changes in the DHFR domain of the DHFR-TS gene (as previously found by other workers); (2) by other changes, possibly affecting the expression of the DHFR-TS gene. The relative importance of these 2 mechanisms in causing resistance in wild populations of P. falciparum is discussed.

Animals↗
PubMed1992

Specificity of affinity-purified Trichinella spiralis antigens.

An affinity-purified fraction (APF) was obtained by passing crude somatic antigens of Trichinella spiralis muscle larvae through an Affi-Gel 10 column coupled with anti-Trichuris suis IgG. The fraction contained seven antigens with molecular weights ranging from 28 to 55 kDa. When tested with antiserum against other common nematodes of pigs from China, the APF was found to be markedly more specific than S3 antigens (prepared by a combination of cell fractionation and differential centrifugation according to Despommier and Lacetti, 1981) and fractions produced by Sephacryl S-200 gel filtration (F1 to F12). When the APF was used in an indirect IgG-enzyme-linked immunosorbent assay (IgG-ELISA) to screen serum samples from 2000 pigs imported from China, a positive rate of 7.5% was obtained. Similar screenings using the crude somatic antigens F1 and S3 gave a large number of cross-reactions and false positive reactions. Positive rates of 48%, 39% and 59.5% respectively were obtained for the three antigens.

Animals↗
PubMed1992

Analysis of a new hepatitis C virus type and its phylogenetic relationship to existing variants.

Sequences obtained in the 5' non-coding region (5'NCR) of hepatitis C virus (HCV) were obtained from Scottish blood donors and compared with previously published HCV sequences. Phylogenetic analysis revealed the existence of three distinct groups of sequences; two of these corresponded to the recently described HCV types 1 and 2 variants, while viral sequences detected in around a third of the blood donors formed a separate phylogenetic group that probably represents infection with a novel virus species. Nucleotide sequences of this latter group differed from all previously published 5'NCR sequence variants by at least 9%. This new virus type also differed considerably from previously published variants in other regions of the viral genome (core, NS-3 and NS-5), with corrected nucleotide distances of 15, 43 and 49% respectively from the prototype HCV-1 sequence. Formal phylogenetic analysis of each of the coding regions confirmed that HCV type 1 variants could be clearly differentiated into regional variants (Far East and U.S.A./European), in contrast to the clearly overlapping geographical distributions of the main HCV types in U.K. blood donors. We discuss the evidence for and against the hypothesis that the three main phylogenetic groups identified in this study represent separate species of HCV.

Amino Acid Sequence↗
PubMed1988

Comparison between standard ELISA and dot-ELISA for serodiagnosis of human trichinosis.

The dot enzyme-linked immunosorbent assay (dot-ELISA) provided rapid and accurate detection of human trichinosis. Comparable sensitivity and specificity of the standard ELISA and dot-ELISA were obtained when sera of 18 confirmed cases of trichinosis were screened. Cross-reactivity was assayed with sera from 31 Chinese subjects with other parasitic infections. Only 2 cross-reactions occurred in the dot-ELISA, compared with 7 in the standard ELISA.

Antigens, Helminth↗
PubMed1987

Four documented cases of eosinophilic meningoencephalitis due to Angiostrongylus cantonensis in Hong Kong.

4 cases of eosinophilic meningoencephalitis in Hong Kong are described. The major clinical features of the patients, who were 2-60 years old, were low grade fever, headache, mild meningeal signs, right facial palsy or hemiplegia. Eosinophilia in the peripheral blood and eosinophilic pleocytosis were prominent. Computerized tomography scans of brain showed a small area of attenuation with surrounding hypodense area; the lesion was resolved 1-2 months after admission to hospital. Electroencephalograms revealed abnormally slow dysrhythmia. Sections of a nematode observed in the brain of a patient who died were identified as those of young adult Angiostrongylus cantonensis. High ELISA titres against the crude antigens of this nematode were also noted in the serum of 3 patients. The disease is probably under-recognized in Hong Kong.

Adult↗