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Biomedical subjects

Y C Chan

Publications and source records attributed to Y C Chan.

At least 91 records · Page 5Linked to original sources
PubMed1994

A shortened dengue IgM capture ELISA using simultaneous incubation of antigen and peroxidase-labeled monoclonal antibody.

A shortened IgM capture ELISA for the detection of dengue IgM antibodies using simultaneous incubation of antigen and peroxidase-labeled monoclonal antibody was described. The shortened two-step assay was compared with the four-step IgM capture ELISA on sera from dengue patients confirmed by the hemagglutination inhibition (HI) test. When paired acute and convalescent sera were tested, the shortened ELISA showed 100% agreement with HI results. It detected dengue IgM antibodies in the acute sera of 66% of patients with a primary dengue infection, 60% of patients with a secondary infection, and 98% of patients with a presumptive secondary infection. When the results of 151 dengue patients were combined, 75% of the acute sera were positive by the shortened IgM capture ELISA.

Journal Article↗
PubMed1994

Comparative analysis of NS3 sequences of temporally separated dengue 3 virus strains isolated from southeast Asia.

By a combination of PCR and direct-cycle sequencing using consensus primers, we analyzed approximately 400-bp fragments within the NS3 genes of twenty-one dengue virus type 3 strains isolated from five neighboring Southeast Asian countries at different time intervals from 1956 to 1992. The majority of base disparities were silent mutations, with few predicted amino acid substitutions, thus emphasizing the strict conservation of the NS3 gene. Phylogenetic trees constructed on the basis of these nucleotide differences revealed distinct but related clusters of strains from the Philippines, Indonesia, and strains from Singapore and Malaysia of the 1970s and early 1980s, while the Thai cluster was relatively more distant. This genetic relationship was compatible with that proposed by other workers who have studied other dengue 3 virus genes such as E, M and prM. However, we observed that the more recent, epidemic-associated dengue 3 strains from Singapore and Malaysia of the late 1980s and early 1990s were more closely related to the Thai cluster, implying their evolution from the latter, and emphasizing the importance of viral spread via increasing travel within the Southeast Asian area and elsewhere. Nucleotide sequence analysis of the NS3 genes of dengue viruses can serve to advance the understanding of the epidemiology and evolution of these viruses.

Amino Acid Sequence↗
PubMed1993

Use of NS3 consensus primers for the polymerase chain reaction amplification and sequencing of dengue viruses and other flaviviruses.

Consensus primers for the polymerase chain reaction were designed based on conserved motifs within the serine protease and RNA helicase domains encoded by the NS 3 genes of dengue and other flaviviruses. Target fragments of 470 bp were amplified on cDNA templates synthesized from RNAs of dengue types 1, 2, 3, and 4, Japanese encephalitis, Kunjin, and yellow fever viruses using random or specific downstream primers. PCR of oligo(dT)-primed cDNAs from Japanese encephalitis and Kunjin viral RNAs did not yield target bands. As few as 10(3) copies of dengue viral RNA could be detected. Direct DNA sequencing of PCR products of reference strains of dengue 2 (NGC), Kunjin (MRM 61C) and yellow fever (17 D) viruses demonstrated complete concurrence with published data. However, 2 nucleotide differences were observed between our data for dengue 3 H87 strain and the published sequence, resulting in a single amino acid disparity. Differences at 21, 16, and 11 nucleotide positions were noted between dengue 1 Hawaii and S 275/90; dengue 4 H 241 and 814669; Japanese encephalitis Nakayama and JaOArS 982 viral strains, culminating in only 4, 1 and 1 amino acid residue differences, respectively. These amino acid disparities occurred outside putative active sites of the enzymatic domains, emphasizing the important role of the NS3 protein in flaviviral replication. This RNA-PCR consensus primer strategy coupled with DNA sequencing represents a valuable tool for the molecular diagnosis and epidemiology of dengue and other flaviviral infections.

Aedes↗
PubMed1993

Alveolar type II cell-fibroblast interactions, synthesis and secretion of surfactant and type I collagen.

During alveolar development and alveolar repair close contacts are established between fibroblasts and lung epithelial cells through gaps in the basement membrane. Using co-culture systems we have investigated whether these close contacts influence synthesis and secretion of the principal surfactant apoprotein (SP-A) by cultured rat lung alveolar type II cells and the synthesis and secretion of type I collagen by fibroblasts. The alveolar type II cells remained cuboidal and grew in colonies on fibroblast feeder layers and on Matrigel-coated cell culture inserts but were progressively more flattened on fixed fibroblast monolayers and plastic. Alveolar type II cells cultured on plastic released almost all their SP-A into the medium by 4 days. Alveolar type II cells cultured on viable fibroblasts or Matrigel-coated inserts above fibroblasts accumulated SP-A in the medium at a constant rate for the first 4 days, and probably recycle SP-A by endocytosis. The amount of mRNA for SP-A was very low after 4 days of culture of alveolar type II cells on plastic, Matrigel-coated inserts or fixed fibroblast monolayers: relatively, the amount of mRNA for SP-A was increased 4-fold after culture of alveolar type II cells on viable fibroblasts. Co-culture of alveolar type II cells with confluent human dermal fibroblasts stimulated by 2- to 3-fold the secretion of collagen type I into the culture medium, even after the fibroblasts' growth had been arrested with mitomycin C. Collagen secretion, by fibroblasts, also was stimulated 2-fold by conditioned medium from alveolar type II cells cultured on Matrigel. The amount of mRNA for type I collagen increased only modestly when fibroblasts were cultured in this conditioned medium. This stimulation of type I collagen secretion diminished as the conditioned medium was diluted out, but at high dilutions further stimulation occurred, indicating that a factor that inhibited collagen secretion also was being diluted out. The conditioned medium contained low levels of IGF-1 and the stimulation of type I collagen secretion was abolished when the conditioned medium was pre-incubated with antibodies to insulin-like growth factor 1 (IGF-1). There are important reciprocal interactions between alveolar type II cells and fibroblasts in co-culture. Direct contacts between alveolar type II cells and fibroblasts appear to have a trophic effect on cultured alveolar type II cells, increasing the levels of mRNA for SP-A. Rat lung alveolar type II cells appear to release a factor (possibly IGF-1) that stimulates type I collagen secretion by fibroblasts.

Animals↗
PubMed1993

Comparative effect of proteins, peptides, and amino acid mixtures on recovery from 70% hepatectomy in rats.

The effects of proteins, peptides, and amino acid mixtures on the recovery from 70% hepatectomy were compared in rats. In Experiment 1, the most suitable time for the comparison was studied. Rats weighing 250 g after 70% hepatectomy were fed 20% casein diet. On day 0,1,2,3,4,6,8,11,15 post-operation, 3-5 rats were killed for the observation of recovery. The recovery time, evaluated by daily food intake, body weight gain, liver weight and compositions, and hematologic values was about 10 days. For the comparison of different nitrogen sources, we estimated that the 6th day after the operation when the animals did not fully recover was suitable. In Experiment 2, rats of each group after hepatectomy were fed one of 14 experimental diets containing protein (3 types), peptides (8 types), or amino acid mixture (3 types) for 5 days and killed on the 6th day after the operation. Daily food intake, body weight gain, liver weight and compositions, and hematologic values were measured and pathological examination was also done. The effect of different nitrogen sources were similar on the recovery after 70% hepatectomy with only a few minor exceptions in rats.

Amino Acids↗
PubMed1993

Improving hospital cost accounting with activity-based costing.

In this article, activity-based costing, an approach that has proved to be an improvement over the conventional costing system in product costing, is introduced. By combining activity-based costing with standard costing, health care administrators can better plan and control the costs of health services provided while ensuring that the organization's bottom line is healthy.

Accounting↗
PubMed1992

A survey of Blastocystis in reptiles.

A total of 28 species of reptiles were investigated for Blastocystis using light microscopy and in vitro culture in biphasic egg slant medium. Blastocystis species were detected in 8 (28.6%) of these 28 species in 3 tortoises (Geochelone elephantopus, G. elegans and G. carbonaria), 3 snakes (Boiga dendrophilla, Python reticulatus and Elaphe radiata), 1 crocodile (Crocodylus porosus) and 1 iguana lizard (Cyclura cornuta). The reptilian Blastocystis appeared to be morphologically similar to B. hominis.

Alligators and Crocodiles↗
PubMed1992

Immunodiagnosis in oral candidiasis. A review.

Detection of anti-Candida antibodies in sera and saliva of patients with oral candidiasis has been regarded as a valuable laboratory technique in the diagnosis of the lesion. However, despite considerable research, the value of candidal immunodiagnosis remains controversial. Conflicting conclusions about the sensitivities and specificities of these techniques as applied to human sera and saliva have appeared. These controversies have arisen because of the use of different antigen preparations and immunologic techniques. For the present, the use of purified cytoplasmic protein antigen of Candida albicans and the ELISA technique seems to be the most reliable laboratory method.

Antibodies, Fungal↗
PubMed1992

Full-length cDNA sequence of dengue type 1 virus (Singapore strain S275/90).

The complete nucleotide sequence and the deduced amino acid sequence of the genome of dengue virus type 1 (Singapore strain S275/90) were determined from cDNA clones. The single-stranded, positive-sense RNA is 10,718 nucleotides in length and contains a single long open reading frame of 10,188 nucleotides encoding a polyprotein of 3396 amino acids. The genomic size and organization were found to be similar to that of other dengue virus serotypes. Both the nucleotide and deduced amino acid sequences were compared with the partial sequence of DEN1 (Nauru Island) and complete sequences of DNE2 (Jamaica), DEN3 (H87), and DEN4 (Dominica) virus genomes.

Amino Acid Sequence↗
PubMed1991

A Blastocystis species from the sea-snake, Lapemis hardwickii (Serpentes: Hydrophiidae).

Observations were made on Blastocystis isolated from the sea-snake, Lapemis hardwickii. Exponential growth of the organism was observed between 2 and 4 days of culture. Vacuolated, amoeboid and granular forms were observed in cultures, similar to B. hominis. The optimal growth temperature for the sea-snake Blastocystis was 24 degrees C compared with 37 degrees C for B. hominis. The karyotypic patterns of B. hominis and the sea-snake Blastocystis were studied in the clamped homogeneous electric field (CHEF) technique and found to be different. Based on the above differences, the sea-snake Blastocystis was designated as Blastocystis lapemi sp. nov.

Animals↗
PubMed1991

Knowledge and concerns of newly diagnosed NIDDM patients in Singapore.

This paper describes the findings of a questionnaire survey of the knowledge and concern of 152 newly diagnosed NIDDM patients presenting at the Government Polyclinic (primary health care centre). This study is part of a project on outpatient diabetic care in Singapore. It was found that 23.7% of the subjects had no education and 51.3% had only primary school education. More than eighty percent of the subjects understood the importance of diet control and weight control in their condition. More than half of the subjects, 61% of males and 52.7% of females were aware that diabetes could not be cured and that once diabetes is controlled, that medication cannot be stopped and that one must still take the medication even if one feels well. Less than 30% knew about the interpretation of blood and urine sugar findings. The two major concerns amongst the subjects were complications and the need for long-term medication. These findings will be incorporated into the next phase of the project where a health education programme for NIDDM patients will be developed and evaluated.

Adult↗
PubMed1991

Sudden unexplained death syndrome--a new manifestation in melioidosis?

The indirect haemagglutination (IHA) test using sensitized turkey erythrocytes and the indirect immunofluorescence assay (IgM-IFA) was confirmed to be sensitive in the detection of a recent or current Pseudomonas pseudomallei infection in 19 culture-confirmed Singapore melioidosis patients. All were found to have antibody titres from 4 to 32768 in the IHA test and 10 to 320 in the IgM-IFA test. When these tests were employed on sera from 16 immigrant Thai construction workers who died of sudden unexplained death syndrome (SUDS) and 73 healthy Thai fellow workers, 93.8% and 68.8% of SUDS cases had IHA titre of greater than or equal to 4 and IgM-IFA titre of greater than or equal to 10 respectively, in contrast to 39.7% and 12.3% found among healthy Thai workers. These data indicate that at the time of death, most of the SUDS patients had an active infection with P. pseudomallei, possibly resulting from reactivation of a latent infection. The aetiological role of P. pseudomallei as the major cause of SUDS is discussed.

Adult↗
PubMed1991

Serodiagnosis of melioidosis in Singapore by the indirect haemagglutination test.

Melioidosis is endemic in Singapore, with diagnosis dependent upon both bacteriological culture and serodiagnosis. Using the polysaccharide (melioidin)-sensitized turkey red cells in the indirect haemagglutination test (IHAT), 20 (100%) of the Pseudomonas pseudomallei culture-positive cases were detectable by the IHAT with titles ranging from 1:16 to 1:32, 768. Eight of these patients who died within a few days after the IHAT was performed had titres ranging from 1:16 to 1:1028. Five culture-negative patients, with clinical symptoms suggestive of melioidosis infection and who responded to treatment with ceftazidime, showed IHA titres between 1:64 and 1:8,192. One hundred and twenty one sera from patients with pneumonia, abscesses, or diabetes mellitus were IHAT negative. The IHAT showed good specificity since negative titres were seen in tests using sera from 2 patients with culture-positive Pseudomonas aeruginosa and 4 patients positive for Legionella. IHAT negative results were obtained from tests of 50 normal blood donors and 50 sewerage workers. Of 683 national servicemen tested, 5 (0.73%) had IHAT titres ranging from 1:16 to 1:128. Unlike hyperendemic areas such as Thailand where interpretation of IHAT is seriously hampered by IHA titres found in one-third to half of the population, serodiagnosis of melioidosis by the sensitive IHAT may be employed in Singapore as a routine procedure since background IHA titres are low.

Hemagglutination Tests↗
PubMed1991

Lymphocyte changes in secondary dengue fever: use of the Technicon H*1 to monitor progress of infection.

Secondary dengue fever as dengue hemorrhagic fever or dengue shock syndrome is a potentially fatal complication of an infection that presents with few clinical signs that help in the diagnosis. Previous workers have reported the value of buffy coat determinations of atypical lymphocytes as an aid to the diagnosis. We report here the use of an automated white blood cell differential counter, the Technicon H*1 in the monitoring of the atypical lymphocyte count as a measure of the progress of the infection in a retrospective study of serial full blood counts in 45 serologically confirmed patients. Technicon H*1 'basophil' and large unstained cell counts and manual atypical lymphocyte counts rose in tandem with the drop in platelets and decreased when the platelets recovered. In a sub-study, the atypical lymphocytes were immunophenotyped and found to be predominantly derived from T lymphocytes. We conclude that the Technicon H*1 is a useful monitor of the activity of the infection in dengue hemorrhagic fever.

Basophils↗
PubMed1990

Passive protection studies in mice with monoclonal antibodies directed against the non-structural protein NS3 of dengue 1 virus.

Antibody-mediated enhancement of dengue virus replication is thought to be a mechanism contributing to the pathogenesis of dengue haemorrhagic fever and dengue shock syndrome. Enhancement is associated with antibodies to structural components of the virus. To circumvent the problem of immune enhancement, studies to identify protective antigens of dengue virus have involved non-structural proteins. Passive and active protection against lethal dengue virus infection in mice have been demonstrated with the non-structural protein NS1. In this study, the dengue virus non-structural protein NS3 was examined in passive protection studies with monoclonal antibodies prepared against NS3 of dengue 1 virus (Hawaiian). Five monoclonal antibodies that were authenticated to be reactive to NS3 were used to immunize 13- to 14-day old mice intraperitoneally. Thereafter, the mice were challenged intracerebrally with 100 LD50 of neurotropic dengue 1 virus and the survival indices of the mice were calculated. Significant decreases in survival indices (P less than 0.05), indicating increases in survival times were observed with four of five monoclonal antibodies tested. Monoclonal antibodies to NS3 of dengue 1 virus are able to increase the survival time of mice challenged with a lethal dose of dengue 1 virus, although the mechanism remains to be defined.

Animals↗
PubMed1990

Prevalence of human papillomavirus types 16 and 18 in cervical carcinomas: a study by dot and Southern blot hybridization and the polymerase chain reaction.

Histologically classified biopsies from 83 women with invasive cervical carcinoma were analyzed by dot blot hybridization for human papillomavirus (HPV) types 16 and 18 infection. Sixty of the 83 (72.3%) were found to contain HPV DNA, of which 43 (51.8%) contained HPV 16 DNA, 12 (14.5%) contained HPV 18 DNA and 5 (6.0%) contained both HPV 16 and 18 DNAs. Southern blot analysis on 65 specimens gave similar results. Of 23 specimens negative by dot blot, 21 were tested by the polymerase chain reaction. Seventeen of the 21 were positive for HPV DNA, of which 13 contained HPV 16 DNA and 4 contained both HPV 16 and 18 DNAs. In all, 95.1% (77/81) were positive for HPV 16 and/or 18 DNA sequences.

Adenocarcinoma↗
PubMed1988

Serological evidence of hantavirus infection in laboratory rats and personnel.

Laboratory-acquired haemorrhagic fever with renal syndrome (HFRS) has been reported in many countries. A serological survey of laboratory white rats and of laboratory personnel for antibodies to hantaviruses was conducted in Singapore. Forty-four per cent (143/329) of rats were seropositive by the indirect immunofluorescent antibody test but none had hantaviral antigens in lung tissues. Two of 74 laboratory personnel were seropositive but neither had a history of clinical illness. The high seropositivity rate among laboratory rats led to their replacement with Hantaan virus-free strains. To eliminate the hazard of laboratory-acquired HFRS, regular serological screening of laboratory rats and replacement of infected animals with seronegative stocks should be implemented. High risk techniques with laboratory rats, which are likely to generate aerosols, should be performed in biological safety cabinets. Serological surveillance of laboratory personnel and reporting of suspected HFRS cases are useful in the early detection of hantavirus infection.

Animals↗