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Biomedical subjects

E H Yap

Publications and source records attributed to E H Yap.

At least 37 records · Page 2Linked to original sources
PubMed1998

Epidemiological surveillance of melioidosis in Singapore.

During the period 1989 to 1996, a total of 372 cases of melioidosis, with 147 deaths, were reported, giving a mean annual incidence rate of 1.7 per 100,000 population and a case-fatality rate of 39.5%. Majority (89%) of the clinical cases were confirmed by culture of Burkholderia pseudomallei, while the others were presumptive cases based on a single blood specimen with an indirect haemaglutination (IHA) antibody titre of > or = 1:16. The highest incidence rate was reported in those aged 45 years and above (5.7 per 100,000 population), males (2.8 per 100,000 population), and Indian ethnic group (3.0 per 100,000 population). Cases were distributed throughout the island all year round. There was no correlation with rainfall. Most of the cases (77.4%) had other concurrent medical conditions, the most common being diabetes mellitus (57.5%). Factors significantly associated with a higher case-fatality rate were age (55 years and above), septicaemia, smoking history and heart or renal failure. The overall case-fatality rate has been declining from 60% in 1989 to 27% in 1996 due to a greater awareness among medical practitioners to diagnose and treat the disease early. The overall seroprevalence of IHA antibody (titre of > or = 1:16) among asymptomatic population groups was 0.2%. B. pseudomallei isolated from clinical specimens were sensitive to imipenem (100%), ceftazidime (99.1%), piperacillin (99.7%), ampicillin-clavulanate (98.5%), minocycline (97.4%), chloramphenicol (94.3%), doxycycline (94.3%) and tetracycline (93.9%). Of 395 samples of soil collected during epidemiological investigation of reported cases, 1.8% were positive for B. pseudomallei.

Adolescent↗
PubMed1997

Description of a Blastocystis species from Rattus norvegicus.

Two isolates (WR1 and WR2) of Blastocystis from laboratory-bred Wistar rats were axenized by a method of colony growth in soft agar combined with antibiotic treatment. The colonies were cultured in Iscove's modified Dulbecco's medium (IMDM) and Bacto agar mixture supplemented with 10% horse serum in the presence of thioglycollate. The cells from the colonies had an ameboid outline with a central body. Large inclusions were seen in the central body of some cells. Some granular forms were also found. In the axenic culture of isolate WR2, about one-third of the organisms were granular forms. Cysts were found in the axenic culture of both isolates. This is the first report of such cyst formation in in vitro culture. The karyotypic patterns of both isolates of the rat Blastocystis were analyzed by pulsed-field gel electrophoresis (PFGE). A total of 13 chromosomal bands were separated, ranging from 1.86 Mb to 295 kb. The karyotypic patterns of the rat Blastocystis were different from those of B. hominis and reptilian Blastocystis. On the basis of the above-mentioned differences, the rat Blastocystis is assigned as B. ratti sp. nov.

Animals↗
PubMed1997

Experimental Blastocystis hominis infection in laboratory mice.

Young (less than 8 weeks old) immunocompetent BALB/c mice became infected with Blastocystis hominis after inoculation of fecal cysts orally and of in vitro axenic-culture forms intracecally. This study confirmed that the fecal cyst was the form responsible for external transmission and that the mode of transmission was by the fecal-oral route. The infection was self-limiting and the infected BALB/c mice appeared normal except that some of them showed weight loss and lethargy. Both vacuolar and granular forms were found in the cecum, but only cyst forms were observed in the colon. Histological examination of the cecum and colon showed intense inflammatory-cell infiltration, edematous lamina propria, and mucosal sloughing. It is apparent that although B. hominis is not invasive, it is capable of causing pathogenesis in BALB/c mice.

Animals↗
PubMed1997

Survival of Blastocystis hominis clones after exposure to a cytotoxic monoclonal antibody.

Our previous studies have shown that monoclonal antibodies (MAbs) to Blastocystis hominis react mainly with carbohydrate epitopes, while 1 MAb (1D5) reacts specifically with a protein of 30.5 kDa. In the present study, 3 monoclonal antibodies (1D5, 1E7 and 4F7) were used in immunogold localization. 1E7 and 4F7 were found to react primarily with the surface coat, while 1D5 was plasma membrane-specific. In the presence of complement, only 1D5 exhibited a cytotoxic effect on B. hominis whereas 1E7 and 4F7 did not, suggesting that the surface coat of B. hominis could serve as an immunological barrier against host antibodies. Using a recently described agar plating method, only 1D5 exhibited significant (P < 0.01) complement-independent cytotoxicity to B. hominis, inhibiting colony growth at low concentrations. Parasites that had been exposed to 1D5 were morphologically smaller than those that were not exposed to this MAb. Colonies that grew in the presence of 1D5 were isolated and grown in liquid medium containing increasing amounts of the cytotoxic MAb. Two clones that grew well in liquid medium containing 1D5 were also able to develop into colonies in soft agar. This study has shown that the 30.5 kDa protein found on the plasma membrane of B. hominis is a functionally important protein and that not all cells within a certain population would be susceptible to the cytotoxic effects of 1D5. These findings suggest that a heterogenous population exists in continuously maintained cultures of B. hominis.

Animals↗
PubMed1996

Recombinant dengue type 1 virus NS5 protein expressed in Escherichia coli exhibits RNA-dependent RNA polymerase activity.

The complete nonstructural NS5 gene of dengue type 1 virus, Singapore strain S275/90 (D1-S275/90) was expressed in Escherichia coli as a glutathione S-transferase (GST) fusion protein (126 kDa). The GST-NS5 fusion protein was purified and the recombinant NS5 protein released from the fusion protein by thrombin cleavage. The recombinant NS5 had a predicted molecular weight of 100 kDa and reacted with antiserum against D1-S275/90 virus in Western blot analysis. The purified recombinant NS5 protein possessed RNA-dependent RNA polymerase activity which was inhibited (>99%) by antibodies against the recombinant NS5 protein. The polymerase product was shown to be a negative-stranded RNA molecule, of template size, which forms a double-stranded complex with the template RNA.

Animals↗
PubMed1996

Axenic culture of reptilian Blastocystis isolates in monophasic medium and speciation by karyotypic typing.

The growth of axenic reptilian isolates of Blastocystis in Iscove's modified Dulbecco's medium (IMDM) was studied and the morphology of the parasite was examined by phase-contrast microscopy. The chromosomal patterns of these reptilian isolates of Blastocystis were examined by pulsed-field gel electrophoresis (PFGE) and compared with those of B. hominis and B. lapemi, a sea snake Blastocystis. IMDM with 10% horse serum supported excellent growth of the reptilian Blastocystis isolates. The parasites from all the isolates were predominantly vacuolar, but multivacuolar and amoeboid forms were also seen. Amoeboid forms with rather elongate pseudopodia were also observed. There were some differences in size, morphology, and growth characteristics in the different reptilian isolates. The karyotypic patterns of the Blastocystis isolates from tortoise, iguana, and python were distinctly different from one another and from those obtained with B. hominis and B. lapemi. On the basis of the above-mentioned differences in chromosomal patterns, the tortoise, iguana, and python isolates are described as new species, viz., B. geocheloni sp. nov. from Geochelone carbonaria (red-footed tortoise), B. cycluri sp. nov. from Cyclura cornuta (rhino iguana), and B. pythoni sp. nov. from Python reticulatus (reticulated python).

Animals↗
PubMed1996

Colony formation of Blastocystis hominis in soft agar.

This is the first description of a method for growing axenized Blastocystis hominis as colonies in petri dishes containing soft agar. Blastocystis cells cultured in two types of agar appeared to show different colonial morphologies as well as differing colony yields. Microscopic examination of the colonies revealed many amoeboid and giant cells. Many cells were also shown to possess thin filament-like structures that appeared to stretch across the central vacuole.

Agar↗
PubMed1996

Observations on the ultrastructure and viability of the cystic stage of Blastocystis hominis from human feces.

This report describes the ultrastructure and viability of cysts of Blastocystis hominis from feces of infected patients. The cysts were round to ovoid, measured 2-5 microns in size, and contained a condensed cytoplasm that had vacuoles of varying sizes, four nuclei, and as many as six cristate mitochondria. The cell wall was rather electron-lucent. Surprisingly, chromatoid-like structures were found in the cytoplasm and nucleus of some of the cysts. These have not previously been reported in Blastocystis. The cysts can survive in water for up to 19 days at normal temperatures but are fragile at extreme temperatures and in common disinfectants.

Animals↗
PubMed1996

Clonal growth of Blastocystis hominis in soft agar with sodium thioglycollate.

The present report describes a method for establishment of colonies of Blastocystis hominis from single cells in soft agar. The percentage of colony-forming efficiency (% CFE = number of colonies grown/number of cells inoculated x 100) for the cultures was greatly improved by the addition of sodium thioglycollate. Five human Blastocystis isolates chosen for this study showed no apparent variation in colonial morphology. Isolated colonies were also successfully grown in liquid medium, providing a means of obtaining large numbers of B. hominis cells that had arisen from a single clone.

Animals↗
PubMed1996

Production and characterization of murine monoclonal antibodies to Blastocystis hominis.

Several hybridomas producing antibodies detected by enzyme-linked immunosorbent assay (ELISA) were established by fusions of mouse myeloma P3.X63.Ag8.U1 with spleen cells from BALB/c mice immunized against an isolate of Blastocystis hominis. Five strongly positive hybrids (6B6, 1D5, 1E7, 4F7 and 4G11) were cloned and all were found to secrete IgM monoclonal antibodies. Four MAbs (6B6, 1E7, 4F7 and 4G11) reacted in immunoblots with a number of B. hominis antigens (mol. wt ranging from 25,000 to 220,000) which were likely to be repeating oligosaccharide epitopes located on glycoproteins, as indicated by pronase and periodate treatment. Another MAb (1D5) reacted with a single antigenic band (mol. wt 30,5000). Similar results were obtained in immunoblots using 4 other B. hominis isolates. Indirect fluorescent-antibody assay (IFA) using MAbs showed 3 patterns of reactivity. 1D5 showed patchy fluorescence, 4F7 showed peripheral fluorescence and 6B6, 1E7 and 4G11 showed bright diffuse fluorescence. These patterns were observed for all 5 human Blastocystis isolates. The MAbs exhibited some cross-reactivity with 2 reptilian Blastocystis isolates but not with Giardia intestinalis, Trichomonas vaginalis or Entamoeba histolytica.

Animals↗
PubMed1996

In vitro response of Blastocystis hominis against traditional Chinese medicine.

This is the first inn vitro study on the activity of 20 kinds of crude extracts of traditional Chinese medicine (TCM) on the intestinal parasite, Blastocystis hominis using the criteria of living cell count (LCC) and living cell rate (LCR). LCC and LCR were applied as observation indicators, the former as a fixed-quantity and the latter as a fixed-quality method. LCR calculated percentage rate of living cells using eosin-brilliant cresyl blue staining which could differentiate between living cells and dying or dead cells. There were five extracts with no inhibitory activity, thirteen with moderate inhibition and two with high inhibition. The crude extracts of Coptis chinensis (CC) and Brucea javanica (BJ) were found to be most active against B. hominis. The active concentration of CC was 100 micrograms/ml. The active concentration of BJ was 500 micrograms/ml. The active concentration of metronidazole (MD) was 10 micrograms/ml and this was taken as an active standard drug for B. hominis.

Animals↗
PubMed1995

Elucidation of the life cycle of the intestinal protozoan Blastocystis hominis.

This paper elucidates the status of the different morphological forms of Blastocystis and reports the existence of thin- and thick-walled cysts in B. hominis on the basis of current experimental evidence. It is suggested that the thin-walled cysts are autoinfectious, leading to multiplication of the organism in the intestinal tract. The thick-walled cysts are responsible for external transmission via the faecal-oral route. A life cycle for B. hominis is postulated on the basis of these findings.

Acridine Orange↗
PubMed1995

Tubulovesicular elements in Blastocystis hominis from the caecum of experimentally-infected rats.

By transmission electron microscopy (TEM), tubulovesicular elements were seen in Blastocystis hominis obtained from the caecum of experimentally-infected rats. These appeared to arise from the peripheral cytoplasm and were rounded, oval or elongate in sections. It is suggested that these elements form a network for transfer of nutrients to the periphery during the process of encystation.

Animals↗
PubMed1995

Serum antibodies to Trichomonas vaginalis in invasive cervical cancer patients.

OBJECTIVE: To evaluate, by seroepidemiology, the possible role of the sexually-transmitted flagellate, Trichomonas vaginalis, in invasive cervical cancer. SUBJECTS AND METHOD: Sera from 121 invasive cervical cancer patients and 242 random age-matched female controls. Antibodies to T. vaginalis were detected by the western blot technique. RESULTS: Antibodies to T. vaginalis were detected in the sera of 41.3% (50/121) of invasive cervical cancer patients compared with only 5.0% (12/242) of female controls. All the reactive sera reacted strongly with the immunogenic surface membrane proteins of T. vaginalis of molecular weights of about 92 and 115 kDa, with variable reactivity to other immunogenic proteins of T. vaginalis. CONCLUSION: The significantly increased relative risk, RR = 3.42 (95% CI = 1.73-6.78), is comparable to the RRs derived in seroepidemiological studies of human papillomavirus, suggesting that T. vaginalis may be even more closely associated with invasive cervical cancer than previously realized.

Adult↗
PubMed1995

Comparison of Pseudomonas pseudomallei from humans, animals, soil and water by restriction endonuclease analysis.

Pseudomonas pseudomallei isolates from 62 human, 17 animal, 3 soil and 3 water samples were examined by genomic DNA digestion with PstI. Five major (RE I, II, III, IV, V) reproducible restriction patterns were observed, with most (56/62) of the human isolates displaying RE I (30/62), II (5/62), III (15/62), IV (4/62), V (2/62), and the animal (16/17), soil (2/3), water (3/3) isolates showing predominantly RE II profiles. Six human and one soil isolates showed patterns different from those of RE I to V. Restriction endonuclease analysis may be applied in epidemiological studies of melioidosis.

Animals↗
PubMed1994

A multiple fission-like mode of asexual reproduction in Blastocystis hominis.

A non-axenic and an axenic isolate of Blastocystis hominis have been induced to form cysts in vitro using an encystation medium. The morphology of the parasite at different time points was observed by scanning electron microscopy. In day-2 cultures the cysts were spherical and had a non-uniform, coarse outer surface around the body. A deep, pore-like opening was seen in some of the parasites. Most of the cysts from day-4 and day-6 cultures ruptured, revealing small, uniformly sized spherical bodies occurring in grape-like clusters. Acridine orange staining confirmed that these bodies were the progeny of Blastocystis hominis. A multiple fission-like reproduction process giving rise to many daughter Blastocystis occurs within the cyst.

Acridine Orange↗