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Biomedical subjects

H C Wong

Publications and source records attributed to H C Wong.

At least 19 recordsLinked to original sources

Characteristics of Hori naevus: a prospective analysis.

BACKGROUND: Acquired, bilateral naevus of Ota-like macules or Hori naevus (HN) is a common dyschromia seen in Orientals. Other than the original report which documented the clinical spectrum in a group of 22 patients, there have not been many epidemiological reports of this condition. OBJECTIVES: To evaluate the epidemiology and clinical characteristics of HN in Asian patients. METHODS: A prospective analysis of 161 patients with HN seen from June 2003 to June 2004 was performed. RESULTS: All 161 patients in the study were women. Patients were Chinese (n = 155), Eurasian (n = 4), Malay (n = 1) and Indian (n = 1). The median age at onset was 30 years. The malar region was the most frequently affected area. Discrete brown macules were the most common early presentation. Confluent slate-grey macules occurred later. Aggravating factors included sun exposure and pregnancy. Sixty-seven patients reported a positive family history. CONCLUSIONS: We report our data on the largest series of HN in the literature so far. Predisposing factors in our study are Chinese race, female sex and positive family history. HN became progressively more confluent and grey over time, suggesting migration from the epidermis to the deeper dermis. More studies are needed to confirm the pathogenesis.

Adolescent↗

Induction of viable but nonculturable state in Vibrio parahaemolyticus and its susceptibility to environmental stresses.

AIMS: This work analysed factors that influence the induction of viable but nonculturable (VBNC) state in the common enteric pathogen, Vibrio parahaemolyticus. The susceptibility of the VBNC cells to environmental stresses was investigated. METHODS AND RESULTS: Bacterium was cultured in tryptic soy broth-3% NaCl medium, shifted to a nutrient-free Morita mineral salt-0.5% NaCl medium (pH 7.8) and further incubated at 4 degrees C in a static state to induce the VBNC state in 28-35 days. The culturability and viability of the cells were monitored by the plate count method and the Bac Light viable count method, respectively. Cells grown at the optimum growth temperature and in the exponential phase better induced the VBNC state than those grown at low temperature and in the stationary phase. Low salinity of the medium crucially and markedly shortened the induction period. The VBNC cells were highly resistant to thermal (42, 47 degrees C), low salinity (0% NaCl), or acid (pH 4.0) inactivation. CONCLUSIONS: Optimal conditions for inducing VBNC V. parahaemolyticus were reported. The increase in resistance of VBNC V. parahaemolyticus to thermal, low salinity and acidic inactivation verified that this state is entered as part of a survival strategy in an adverse environment. SIGNIFICANCE AND IMPACT OF THE STUDY: The methods for inducing VBNC V. parahaemolyticus in a markedly short time will facilitate further physiological and pathological study. The enhanced stress resistance of the VBNC cells should attract attention to the increased risk presented by this pathogen in food.

Colony Count, Microbial↗

Effects of heat, acid, and freeze-thaw challenges on survival of starved Vibrio parahaemolyticus in minimal salt medium, tryptic soy broth, and filtered oyster homogenate medium.

Vibrio parahaemolyticus is a ubiquitous gram-negative enteropathogenic bacterium. To evaluate the risk of stress-adapted V. parahaemolyticus cells in food, we investigated the survivability of starvation-adapted and starvation-low salinity-adapted cells of this pathogen in different media against different stresses. Logarithmically grown bacterial cells were starved at 25 degrees C in a minimal salt medium with 0.5 or 3.0% NaCl for 24 h. Resistances against challenges of heat, acid, and freeze-thaw treatment exhibited by the starvation-adapted cells were similar to those exhibited by the starvation-low salinity-adapted cells but substantially higher than those of the unadapted control cells. The increased stress resistance of the adapted cells against freeze-thaw challenge was lower in tryptic soy broth than in the starving medium. Resistance of the adapted bacteria against heat and freeze-thaw treatment was completely eliminated in filter-sterilized oyster homogenate medium. Practically, these results help to assess the risk of stress-adapted V. parahaemolyticus in food.

Adaptation, Physiological↗

Involvement of spinal calcitonin gene-related peptide in the development of acute visceral hyperalgesia in the rat.

This study aimed to characterize the role of the neuropeptide calcitonin gene-related peptide (CGRP) in the development of mechanically induced visceral hyperalgesia. Tonic colorectal distension (CRD) was performed in fasted, conscious male Sprague-Dawley rats. The visceromotor reflex associated with noxious CRD was determined as the number of contractions during each of two consecutive tonic distensions (10 min at 60 mmHg), which were separated by a series of phasic distensions (repeated 15-s distensions to 80 mmHg at 30-s intervals). The effect of the CGRP receptor antagonist h-CGRP8-37 given intrathecally (i.t.) (0.03-3 nmol rat-1) or intravenously (i.v.) (20 microg kg-1 bodyweight [bw]) on the visceromotor response was evaluated. The dose for i.v. administration was chosen based on previous results from similar studies. In addition, the effect of a CGRP monoclonal antibody (6 mg kg-1 bw) given intravenously was evaluated. Compared to the baseline response, a significant increase in the number of abdominal contractions was observed during the second tonic distension. The i.t. application of h-CGRP8-37 dose-dependently reduced the numbers of abdominal contractions both during the first and the second tonic distension period, with a maximum effect observed at a peptide concentration of 3 nmol. Intravenous administration of h-CGRP8-37 or of the CGRP antiserum produced a small reduction of the visceromotor response induced by the second tonic distension and had no effect on colonic compliance. The development of mechanically induced colorectal hyperalgesia by repeated tonic distension involves the spinal release of CGRP, while peripheral release of CGRP plays only a minor role.

Acute Disease↗

Physician use of complementary and alternative medicine (CAM) literature.

OBJECTIVE: To document the interest of complementary and alternative medicine (CAM) literature use among physicians. DESIGN: A retrospective study. SETTING: A medical library service. MAIN OUTCOME MEASURES: Analysis of photocopy requests from published lists of reference in a two-year period by physicians in the province of British Columbia, Canada. RESULTS: The average number of requests per article was 18.7 among CAM literature and 7.1 among non-CAM literature. Ten of the 15 most frequently requested articles were CAM papers. Review articles on herbal medicine demonstrated the most use. CONCLUSIONS: There is a definite interest in CAM literature among a specific group of Canadian physicians. Use of scientifically credible literature in peer-reviewed journals will be helpful for patient care, teaching, and research in this field.

British Columbia↗

Monoclonal antibody to rat galanin: production, characterization, and in vivo immunoneutralization activity.

A monoclonal antibody (MAb) to galanin was prepared by cell fusion of myeloma Fox-NY and spleen cells from Robertsonian mice immunized with rat galanin. Hybridomas producing high-affinity antibodies were cloned in pristine-primed Balb/c mice. The antibody was purified by affinity chromatography and concentrated to 12 mg IgG/mL by dialysis. Immunoreactivity of the antibody was screened by radioimmunoassay. Ascites fluid contained approximately 10 mg/mL IgG that belong to the subclass of IgG2a as determined by enzyme-linked immunoadsorbent assay (ELISA). The titer of this IgG2a antibody entitled #G65G was 1:10,000 and the ID50 for rat galanin was 1000 fmol/mL as determined by liquid phase radioimmunoassay. Immunohistochemistry showed that this galanin MAb stains densed, beaded processes distributed to the enteric plexuses, where they appear to encircle neuronal cell bodies, to the muscle layer, where they are particularly abundant in the circular muscle layer and in the deep muscular layer, and to the mucosa. In vivo capacity of immunoneutralization by this antibody was tested in male Sprague-Dawley rats fasted for 24 h and anesthetized with urethane. Systemic injection of protein A purified galanin antibody (6 mg/rat) decreased by 70% of the inhibitory effect of intravenous galanin (2 nmol/kg/h i.v.) on gastric acid secretion induced by intracisternal TRH analog. These results show that galanin antibody #G65G is useful for in vivo immunoneutralization of galanin effects and is a valuable tool for immunohistochemical localization of galanin in gastrointestinal tissues.

Animals↗

Evaluation of typing of vibrio parahaemolyticus by three PCR methods using specific primers.

Vibrio parahaemolyticus is a halophilic bacterium frequently involved in human outbreaks of seafood-associated gastroenteritis. For epidemiological purposes, different molecular typing methods, such as pulsed-field gel electrophoresis (PFGE) or ribotyping, have been developed for this pathogen; however, these methods are mostly labor-intensive and time-consuming. In this work, we designed and evaluated three rapid PCR typing methods for this pathogen using primers designed on the basis of the following specific sequences: conserved ribosomal gene spacer sequence (RS), repetitive extragenic palindromic sequence (REP), and enterobacterial repetitive intergenic consensus sequence (ERIC). Typing patterns and clustering analysis indicated that these methods apparently differentiated V. parahaemolyticus strains from reference strains of interspecific Escherichia coli, V. cholerae, and V. vulnificus and were also valuable in subspecies typing of this pathogen. Forty domestic strains of V. parahaemolyticus, representing a wide range of PFGE patterns, were grouped into 15, 27, and 27 patterns, with discrimination indexes of 0.91, 0.97, and 0.98, by RS-, REP-, and ERIC-PCR, respectively. The discriminative abilities of these PCR methods closely approached or even exceeded those of PFGE and ribotyping. REP-PCR is preferable to ERIC-PCR because of the greater reproducibility of its fingerprints, while RS-PCR may be a practical method because it generates fewer amplification bands and patterns than the alternatives.

Bacterial Typing Techniques↗

Structural basis of the recognition of the dishevelled DEP domain in the Wnt signaling pathway.

The DEP domain of Dishevelled (Dvl) proteins transduces signals to effector proteins downstream of Dvl in the Wnt pathway. Here we report that DEP-containing mutants inhibit Wnt-induced, but not Dvl-induced, activation of the transcription factor Lef-1. This inhibitory effect is weakened by a K434M mutation. Nuclear magnetic resonance spectroscopy revealed that the DEP domain of mouse Dvl1 comprises a three-helix bundle, a beta-hairpin 'arm' and two short beta-strands at the C-terminal region. Lys 434 is located at the tip of the beta-hairpin 'arm'. Based on our findings, we conclude that DEP interacts with regulators upstream of Dvl via a strong electric dipole on the molecule's surface created by Lys 434, Asp 445 and Asp 448; the electric dipole and the putative membrane binding site are at two different locations.

3T3 Cells↗

Characteristics of Vibrio parahaemolyticus O3:K6 from Asia.

A variety of serovars of the food-borne pathogen Vibrio parahaemolyticus normally cause infection. Since 1996, the O3:K6 strains of this pathogen have caused pandemics in many Asian countries, including Taiwan. For a better understanding of these pandemic strains, the recently isolated clinical O3:K6 strains from India, Japan, Korea, and Taiwan were examined in terms of pulsed-field gel electrophoresis (PFGE) typing and other biological characteristics. After PFGE and cluster analysis, all the O3:K6 strains were grouped into two unrelated groups. The recently isolated O3:K6 strains were all in one group, consisting of eight closely related patterns, with I1(81%) and I5(13%) being the most frequent patterns. Pattern I1 was the major one for strains from Japan, Korea, and Taiwan. All recently isolated O3:K6 strains carried the thermostable direct hemolysin (tdh) gene. No significant difference was observed between recently isolated O3:K6 strains and either non-O3:K6 reference strains or old O3:K6 strains isolated before 1996 with respect to antibiotic susceptibility, the level of thermostable direct hemolysin, and the susceptibility to environmental stresses. Results in this study confirmed that the recently isolated O3:K6 strains of V. parahaemolyticus are genetically close to each other, while the other biological traits examined were usually strain dependent, and no unique trait was found in the recently isolated O3:K6 strains.

Anti-Bacterial Agents↗

Pandemic spread of an O3:K6 clone of Vibrio parahaemolyticus and emergence of related strains evidenced by arbitrarily primed PCR and toxRS sequence analyses.

Vibrio parahaemolyticus O3:K6 strains responsible for the increase in the number of cases of diarrhea in Calcutta, India, beginning in February 1996 and those isolated from Southeast Asian travelers beginning in 1995 were shown to belong to a unique clone characterized by possession of the tdh gene but not the trh gene and by unique arbitrarily primed PCR (AP-PCR) profiles (J. Okuda, M. Ishibashi, E. Hayakawa, T. Nishino, Y. Takeda, A. K. Mukhopadhyay, S. Garg, S. K. Bhattacharya, G. B. Nair, and M. Nishibuchi, J. Clin. Microbiol. 35:3150-3155, 1997). Evidence supporting a hypothesis that this clone emerged only recently and is spreading to many countries was obtained in this study. Of 227 strains isolated in a hospital in Bangladesh between 1977 and 1998, only 22 strains isolated between 1996 and 1998 belonged to the new O3:K6 clone (defined by the serovar, the tdh and trh typing, and AP-PCR profiles). The O3:K6 strains isolated from clinical sources in Taiwan, Laos, Japan, Thailand, Korea, and the United States between 1997 and 1998 were also shown to belong to the new O3:K6 clone. The clonality of the new O3:K6 strains was also confirmed by analysis of the toxRS sequence, which has been shown to be useful for phylogenetic analysis of the members of the genus Vibrio. The toxRS sequences of the representative strains of the new O3:K6 clone differed from those of the O3:K6 strains isolated before 1995 at least at 7 base positions within a 1,346-bp region. A new PCR method targeted to 2 of the base positions unique to the new O3:K6 clone was developed. This PCR method could clearly differentiate all 172 strains belonging to the new O3:K6 clone from other O3:K6 strains isolated earlier. One hundred sixty-six strains belonging to 28 serovars other than O3:K6 were also examined by the new PCR method. The tdh-positive and trh-lacking strains that belonged to the O4:K68 and O1:K untypeable serovars and were isolated in three countries and from international travelers beginning in 1997 gave positive results. The AP-PCR profiles of these strains were nearly identical to those of the new O3:K6 clone, and their toxRS sequences were 100% identical to that of the new O3:K6 clone. The results suggest that these strains may have diverged from the new O3:K6 clone by alteration of the O:K antigens. In conclusion, this study presents strong evidence for the first pandemicity in the history of V. parahaemolyticus and reports a novel toxRS-targeted PCR method that will be useful in epidemiological investigation of the cases associated with the current pandemic spread.

Bacterial Proteins↗

Characterization of Vibrio parahaemolyticus isolates obtained from foodborne illness outbreaks during 1992 through 1995 in Taiwan.

Vibrio parahaemolyticus is an important foodborne pathogen in Taiwan and many other Asian countries. A total of 371 isolates of V. parahaemolyticus collected from patients involved in foodborne illness outbreaks in Taiwan from 1992 to 1995 were characterized. These isolates had typical biochemical characteristics and only 4% were urease positive. The most frequently isolated serovars were O5:K15 (18.5%), O4:K8 (16.2%), O3:K29 (12.5%), O1:K56 (8.3%), O2:K3 (6.5%), and O4:K12 (6.0%). Most of the isolates were susceptible to nalidixic acid, tetracycline, tobramycin, cephalothin, and gentamicin. About 10% of the isolates were resistant to seven or more antibiotics. Approximately 92.4% of these V. parahaemolyticus showed beta-hemolysis on Wagatsuma blood agar plate and approximately 62.1% of these isolates exhibited detectable amounts of thermostable direct hemolysin. Most of the isolates examined exhibited two copies of tdh genes on the 1.3- and 2.5-kb HindIII-digested chromosome fragments with several variations on other fragments. A pulsed-field gel electrophoresis (PFGE) subspecies typing scheme was used to analyze these domestic isolates and the O3:K6 strains from Japan, Korea, and Taiwan. Fifty seven patterns were differentiated with A, B, C, E, and H being the major domestic types (cumulatively 76% of isolates), while O3:K6 strains (PFGE type I), abruptly occurring since 1996, were genetically distant from the major domestic types.

Animals↗

Incidence of highly genetically diversified Vibrio parahaemolyticus in seafood imported from Asian countries.

Vibrio parahaemolyticus is an important pathogen of humans and aquacultured animals, especially in Asian countries. In this study, we examined 686 samples of seafood imported from Hong Kong, Indonesia, Thailand and Vietnam for V. parahaemolyticus. V. parahaemolyticus was recovered from 315 (45.9%) samples. The incidence of V. parahaemolyticus in products from Hong Kong and Thailand was markedly higher than the incidence in products from Indonesia and Vietnam. The incidence rates in shrimp, crab, snail, lobster, sand crab, fish and crawfish were 75.8, 73.3, 44.3, 44.1, 32.5, 29.3 and 21.1%, respectively. None of the isolates possessed the hemolysin genes (tdh, trh). The chromosomal DNA of 121 randomly selected imported isolates and three local environmental strains was digested with SfiI and analyzed by pulsed-field gel electrophoresis (PFGE). These isolates were then grouped into 96 different but mostly unique PFGE patterns. After hierarchical cluster analysis, these patterns could be arbitrarily grouped into twenty-two PFGE types (type A to V). A wide range of PFGE types were identified in isolates from different origin. Moreover, the PFGE types were not specifically associated with the origin or kind of seafood. These results reveal the high genetic diversity in V. parahaemolyticus isolated from seafood.

Animals↗