Identification of human and bovine rotavirus serotypes by polymerase chain reaction.

The use of the polymerase chain reaction (PCR) for identifying serotypes of human and bovine rotaviruses was examined. In the identification of 115 human rotavirus samples in stools, results with PCR showed excellent agreement with results of serotyping by an enzyme-linked immunosorbent assay (ELISA) using serotype-specific monoclonal antibodies. Furthermore, the PCR showed a much higher sensitivity (93%) than the ELISA test (82.6%). The PCR method could also be applied for identifying the serotype of bovine rotaviruses.     

Molecular epidemiology of enterovirus 71 infection in the Western Pacific Region

BACKGROUND: Recently, there have been large outbreaks of hand, foot and mouth disease (HFMD) mainly caused by enterovirus 71 (EV71) associated with severe neurological diseases in the Western Pacific Region (WPR). To monitor the realtime trend of EV71 transmission throughout the WPR, the authors conducted a molecular epidemiological analysis of EV71 infection. METHODS: Viruses were isolated from clinical samples from patients with HFMD or those with neurological complications. The EV71 isolates were identified by microneutralization assay. The VP4 and/or VP1 regions of recent EV71 isolates were sequenced and subjected to phylogenetic analysis using reference EV71 strains. RESULTS: The phylogenetic analysis of EV71 isolates from the WPR revealed two major genogroups, B and C, based on the nucleotide sequence alignment of the VP1 or VP4 region. These two major genogroups were further divided into subgenogroups, B1, B2, B3, and B4 and C1, C2, C3 and C4, respectively. CONCLUSIONS: The molecular epidemiological analyses of recent and previous EV71 isolates in the WPR indicated that two major genogroups of EV71 are co-circulating in Australia, Malaysia, Singapore, Taiwan and Japan. Recent EV71 isolates in Mainland China constitute a new distinct genetic cluster, subgenogroup C4. Two major lineages of EV71 are the major causative agents of the present HFMD epidemics in the WPR and both are considered to be neurovirulent.     

Antibacterial caged-tetraprenylated xanthones from the stem bark of Garcinia scortechinii

Five new caged-tetraprenylated xanthones, scortechinones L - P (1-5), together with six known scortechinones (A, B, D, F, I and J) and one known xanthone, 4 ',5 '-dihydro-1,5-dihydroxy-6 ',6 '-dimethylpyrano(2 ',3 ':6,7)-4 ',4 ',5 '-trimethylfurano(2 ',3 ':3,4)- xanthone, were isolated from the crude methanol extract of the stem bark of Garcinia scortechinii. The structures were elucidated by analysis of spectroscopic data and comparison of the NMR data with those reported previously. The antibacterial activity of all caged-polyprenylated xanthones, isolated from the latex and stem bark of G. scortechinii, was evaluated. Scortechinone B (6) exhibited significant antibacterial activity against a methicillin-resistant Staphylococcus aureus strain with an MIC value of 2 microg/mL. From the MIC values, some structure-antibacterial activity relationships were established.     

Serological and genetic characterization of bovine rotaviruses in Thailand by ELISA and RNA-RNA hybridization: detection of numerous non-serotype 6 strains

A total of 62 fecal specimens positive for rotavirus were collected from diarrheic cows in Thailand in 1988 and 1989. The antigenic properties of rotaviruses in stool were examined by enzyme-liked immunosorbent assays using specific monoclonal antibodies directed at VP4, VP6 or VP7: all the bovine rotavirus strains were determined as subgroup I; none of the strains were reactive with serotype 6-specific monoclonal antibody; and different reactivities of the bovine strains with two anti-VP4 monoclonal antibodies were observed. Polyacrylamide gel electrophoresis of viral RNA exhibited three different RNA electropherotypes. In RNA-RNA hybridization experiments using cell culture-adapted three strains as well as a reference bovine strain (NCDV), RNA from the Thai bovine strains showed very low homology to that from NCDV; only three or four RNA segments were hybridized between the RNAs from Thai samples and NCDV. These results suggested that bovine rotaviruses isolated in Thailand are serologically and genetically distinct from a reference serotype 6 bovine strain, NCDV.     

Distinct yearly change of serotype distribution of human rotavirus in Thailand as determined by ELISA and PCR.

A total of 241 group A rotavirus-positive stool samples collected from diarrhoeic patients in Thailand between July 1988 and June 1991 were characterized for their serotypes by enzyme-linked immunosorbent assay (ELISA) using serotype-specific monoclonal antibodies and by a polymerase chain reaction (PCR). In July 1988-June 1989, serotype 1 was the most prevalent (63.4%), followed by serotype 4 (11.0%) and serotype 2 (8.5%). In July 1989-June 1990, 59.8% were serotype 1, 24.3% were serotype 2, and 6.1% were serotype 3. In contrast, in July 1990-June 1991, serotype 3 was detected in the highest frequency (40.5%), 29.9% were serotype 1, and 27.3% were serotype 2. Thus, a distinct yearly change of serotype distribution of rotavirus in Thailand was observed in the three consecutive years. In particular, it was of note that the prevalence of serotype 3 greatly increased, in contrast to the previous studies in which almost no serotype 3 rotaviruses were detected in the years 1983-8 in Thailand.     

A long‐term survey on the distribution of the human rotavirus G type in Thailand

The distribution of the G type of human rotavirus was surveyed in Thailand between July 1993 and June 2007. A significant yearly change in the distribution of the G type distribution was found. From 1993–1994 to 1998–1999, the G1 type was the most dominant. In 1999–2000, G9 began to appear at a high frequency. In 2000–2001, 2001–2002, and 2002–2003, G9 was very common. In 2003–2004, G1 became the most prevalent type again, and since then it has been detected at the highest frequency. G12 strains, which were first detected in 1998–1999, were also found in 2004–2005 and 2006–2007. The G4 and G3 types were moderately prevalent in 2001–2002 and 2004–2005, respectively. Nucleotide sequence analysis of the VP7 genes of the G9 and G12 strains which reemerged in Thailand showed that they were each similar to the contemporary strains in other countries. J. Med. Virol. 82:157–163, 2010. © 2009 Wiley‐Liss, Inc.     

Improvement of function and morphology of tumor necrosis factor-alpha treated endothelial cells with 17-beta estradiol: a preliminary study for a feasible simple model for atherosclerosis.

BACKGROUND: Dysfunction of endothelial cells (EC) to produce endothelial nitric oxide synthase (eNOS) by tumor necrosis factor-alpha (TNF-alpha) causes critical features of vascular inflammation associated with several disease states (eg, atherosclerosis including increased platelet aggregation and adhesion on EC, elevated adhesion molecules and enhanced inflammatory cells binding to EC). 17-beta estradiol (E2) can stimulate eNOS production and improve the critical features of atherosclerosis. Using TNF-alpha and E2, we attempted to develop an in vitro vascular model for studying atherosclerosis. METHODS AND RESULTS: Human umbilical vein endothelial cells (HUVEC) grown in transwells were cocultured with smooth muscle cells in a 24-well plate to mimic the major components of the vascular wall. The model was incubated with TNF-alpha (10 ng/ml) for 12 h, prior exposed to E2 (100 pg/ml) for 6-12 h, then investigated by transmission and scanning electron microscopes. The result indicated recovered morphology with good tight junction, and decreased platelet adhesion was noted in defective HUVEC after E2 treatment. CONCLUSION: 17-beta estradiol was represented as an antiatherosclerogenic agent to demonstrate feasibility of the model. Although our finding focused only on the endothelium, this would be the basis for our future studies to develop ex vivo continuous perfusion of human vessel segments for a further atherosclerosis study.     

Therapeutic effect of subcurative dose praziquantel on Schistosoma mansoni infected mice and resistance to challenge infection after treatment

The therapeutic effect of a subcurative dosage of praziquantel (PZQ) on Schistosoma mansoni infected mice and resistance to challenged worm infection after treatment were assessed and compared with conventional treatment using a curative dosage of PZQ. S. mansoni infected mice were treated with PZQ at a curative dosage (600 mg kg(-1)) or a subcurative dosage (300 mg kg(-1)) at 9 weeks after infection. Untreated mice and non-infected mice were added as controls. The therapeutic effect of the drug was evaluated in terms of the mortality of mice after treatment, and the parasitological and pathological findings in mice sacrificed at 1 week, 1 month, or 3 months after treatment. Another sample of mice was not killed but challenged with S. mansoni cercariae at 1 week, 1 month, or 3 months after treatment. Resistance to re-infection was evaluated by the extent of challenged worm reduction. In conclusion, there was no significant difference in mortality, or parasitological and pathological findings between mice treated with PZQ at the two dosages. However, resistance to challenged worm infection was more sustained in the group treated with subcurative dose PZQ, especially at 3 months after treatment.     

Development of an antigen ELISA to detect sapovirus in clinical stool specimens

Summary. Human sapovirus (SaV) strains are etiological agents of mild and/or acute gastroenteritis in children and adults. In this study, we describe the development of a novel antigen enzyme-linked immunosorbent assay (ELISA) detection system that was based on hyperimmune rabbit and guinea pig antisera raised against SaV genogroup I (GI) virus-like particles. The ELISA had 100% specificity, and sensitivities of 60% and 25% when compared to single-round PCR and nested PCR, respectively. Our results have shown the ELISA was useful in detecting SaV GI antigens in clinical stool specimens collected two days after the onset of illness.