Optimization and head-to-head comparison of MISSR-PCR, ERIC-PCR, RAPD and 16S rRNA evolutionary clock for the genotyping of Vibrio cholerae isolated in China.

Purpose: To establish a new genotyping method for Vibrio cholerae and compare it with other methods. Materials and Methods: In the current study, a modified inter simple sequence repeat-polymerase chain reaction (MISSR-PCR) system was developed via several rounds of optimisation. Comparison study was then conducted between MISSR-PCR and three other methods, including enterobacterial repetitive intergenic consensus sequences-based PCR (ERIC-PCR), randomly amplified polymorphic DNA (RAPD) and 16S rRNA evolutionary clock, for the detection and genetic tracing of Vibrio cholerae isolated from seafood in China. Result: The results indicated that the MISSRPCR system could generate the highest polymorphic fingerprinting map in a single round PCR and showed the best discriminatory ability for Vibrio cholerae genotyping by clearly separating toxigenic/nontoxigenic strains, local/foreign strains, and O1/O139/non-O1/non-O139 serogroup strains, comparing to ERIC-PCR, RAPD and 16S rRNA evolutionary clock. Moreover, the MISSR-PCR is superior to previously described traditional simple sequence repeat based PCR method on genotyping by more clearly separating different clusters. Conclusion: To the best of our knowledge, this is the first head-to-head comparison of four detection and genotyping methods for Vibrio cholerae. The MISSRPCR system established here could serve as a simple, quick, reliable and cost-effective tool for the genotyping and epidemiological study.

Effect of vitamin D therapy in addition to amitriptyline on migraine attacks in pediatric patients

The purpose of this study was to investigate the effect of supplementary vitamin D therapy in addition to amitriptyline on the frequency of migraine attacks in pediatric migraine patients. Fifty-three children 8-16 years of age and diagnosed with migraine following the International Headache Society 2005 definition, which includes childhood criteria, were enrolled. Patients were classified into four groups on the basis of their 25-hydroxyvitamin D [25(OH)D] levels. Group 1 had normal 25(OH)D levels and received amitriptyline therapy alone; group 2 had normal 25(OH)D levels and received vitamin D supplementation (400 IU/day) plus amitriptyline; group 3 had mildly deficient 25(OH)D levels and received amitriptyline plus vitamin D (800 IU/day); and group 4 had severely deficient 25(OH)D levels and was given amitriptyline plus vitamin D (5000 IU/day). All groups were monitored for 6 months, and the number of migraine attacks before and during treatment was determined. Calcium, phosphorus alkaline phosphatase, parathormone, and 25(OH)D levels were also determined before and during treatment. Results were compared between the groups. Data obtained from the groups were analyzed using one-way analysis of variance. The number of pretreatment attacks in groups 1 to 4 was 7±0.12, 6.8±0.2, 7.3±0.4, and 7.2±0.3 for 6 months, respectively (all P>0.05). The number of attacks during treatment was 3±0.25, 1.76±0.37 (P<0.05), 2.14±0.29 (P<0.05), and 1.15±0.15 (P<0.05), respectively. No statistically significant differences in calcium, phosphorus, alkaline phosphatase, or parathormone levels were observed (P>0.05). Vitamin D given in addition to anti-migraine treatment reduced the number of migraine attacks.

A rare cause of Rhabdomyolysis from Brucella: a case report / Una causa rara de Rabdomiolisis a partir de Brucella: un reporte de caso

Rhabdomyolysis is caused by myocyte necrosis, which results in the release of muscular cell contents into the circulation and extracellular fluid. We present a case of rhabdomyolysis due to brucella infection without any complications. Following the treatment for brucella, creatinine kinase level was significantly reduced. Rhabdomyolysis associated with brucella is rare in children.

La rabdomiolisis es causada por necrosis del miocito, que resulta de la liberación de los contenidos de la célula muscular en la circulación y el líquido extracelular. Presentamos un caso de rabdomiolisis debido a infección por brucella sin complicaciones. Tras el tratamiento aplicado por brucella, se redujo significativamente el nivel de creatinina kinasa. La rabdomiólisis asociada a brucella es rara en niños.

Growth inhibitory effect and Chk1-dependent signaling involved in G2/M arrest on human gastric cancer cells induced by diallyl disulfide

Diallyl disulfide (DADS) inhibits growth and induces cell cycle G2/M arrest in human gastric cancer MGC803 cells. In this study, 15 mg/L DADS exerted similar effects on growth and cell cycle arrest in human gastric cancer BGC823 cells. Due to the importance of cell cycle redistribution in DADS-mediated anti-carcinogenic effects, we investigated the role of checkpoint kinases (Chk1 and Chk2) during DADS-induced cell cycle arrest. We hypothesized that DADS could mediate G2/M phase arrest through either Chk1 or Chk2 signal transduction pathways. We demonstrated that DADS induced the accumulation of phosphorylated Chk1, but not of Chk2, and that DADS down-regulated Cdc25C and cyclin B1. The expression of mRNA and total protein for Chkl and Chk2 was unchanged. Chk1 is specifically phosphorylated by ATR (ATM-RAD3-related gene). Western blot analysis showed that phospho-ATR was activated by DADS. Taken together, these data suggest that cell cycle G2/M arrest, which was associated with accumulation of the phosphorylated forms of Chk1, but not of Chk2, was involved in the growth inhibition induced by DADS in the human gastric cancer cell line BGC823. Furthermore, the DADS-induced G2/M checkpoint response is mediated by Chk1 signaling through ATR/Chk1/Cdc25C/cyclin B1, and is independent of Chk2.

Varicella epidemiology and cost-effectiveness analysis of universal varicella vaccination program in Taiwan.

Varicella zoster virus is highly contagious and affects people worldwide. In this study, we collected local epidemiological data and evaluated the cost-effectiveness of varicella vaccination program in Taiwan. To examine the economical consequences of universal vaccination, a model of the incidence and the associated costs in a hypothetical cohort was created each year for 30 years. The incidence increased sharply after the infancy and peaked in children aged 5 years. The hospitalization rate among cases was the highest in infants, followed by adults 30 to 44 years old. The benefit-cost analysis showed that one dollar invested in the program would cost extra 46 cents in direct medical expense, but would save extra 45 cents considering the societal expenses. Substantial economical benefits can occur due to the averted unproductive days for parents. Sensitive surveillance of both varicella and zoster is essential in countries that have implemented or are about to implement varicella vaccination.

Prevalence of dengue fever and dengue hemorrhagic fever in Hospital Tengku Ampuan Rahimah, Klang, Selangor, Malaysia.

Dengue fever and dengue hemorrhagic fever have been known to be endemic and reportable diseases in Malaysia since 1971. Major outbreaks occurred in 1973, 1982 and in 1998. For the past few decades until now. many studies have been performed to investigate the importance of these two diseases in Malaysia. A retrospective study was carried out in Hospital Tengku Ampuan Rahimah Klang to find the prevalence of these diseases. The data was collected from the record department of this hospital starting from the year 1999 until 2003 (5 years). A total of 6,577 cases of dengue fever and 857 cases of dengue hemorrhagic fever were reported. From the year 2000 onwards, cases of dengue fever had increased tremendously. However for the year 2001, there was a slight decrease in the reported cases. Most cases occurred in 2003, increasing from 674 in 1999 to 2,813 in 2003. Highest incidence was seen in Malay males more than 12 years of age. However, the cases of dengue hemorrhagic fever declined tremendously throughout the years. Most cases occurred in 1999 with 674 cases, then declining to only one in the year 2001 before it increased to 60 and 72 in the years 2002 and 2003, respectively. Most cases occurred in patients above 12 years old, the majority of which were Malay males.

A seroepidemiology study of varicella among children aged 0-12 years in Taiwan.

The epidemiology pattern of varicella appears to vary among regions with different climates, population densities, and degrees of development. This study investigated the age-specific varicella zoster virus (VZV) seroprevalence in children aged 0 to 12 years in Taiwan and compared these seroprevalences between free and private vaccination areas. Residual sera were collected from 13 hospitals with 1,401 valid samples. Immunoglobulin G antibodies to VZV were measured by enzyme-linked immunosorbent assay. Parents of 656 children answered questions about the varicella incidence and varicella vaccination history of their children. In the 8-12 year-olds, the seroprevance ranged between 88.0-93.8% in northern, central, and eastern, while it was only 76.1% in southern Taiwan. The seroprevalence of children 0-5 years old were significantly different between free and private vaccination areas. Seropositive children who reported no history of varicella or receiving varicella vaccine accounted for 26.1-59.3% of the total positive cases. Our findings suggest the possible effects of climate, geographical conditions, and lifestyle on the seroepidemiology of VZV in Taiwan. The efforts of implementing a varicella vaccination program in Taiwan should focus on reaching high levels of coverage.

Allergens of Bipolaris species.

Skin prick tests done previously revealed a significantly higher percentage of sensitization to an extract of Bipolaris sp. among atopic individuals (34/147, 23.1%) compared to non-atopic individuals. Bipolaris-specific IgE levels were quantified in sera from a representative group of 38 individuals using the Fluorescence Allergosorbent Test (FAST). Result obtained by FAST were found to be comparable to the skin prick test results (r2 = 0.60, p < 0.001 for IgE levels vs wheal sizes; r2 = 0.44, p < 0.001 for IgE levels vs erythema sizes). Characterisation of the extract's allergenic component by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) showed 28 protein bands with molecular weights (MW) ranging from 11 kDa to above 100 kDa. Immunoblotting with sera of 10 Bipolaris-sensitive (skin prick test, 3 +) individuals showed that Bipolaris spore extract contained at least 4 IgE binding proteins (MW 11-13 kDa, 16-17 kDa, 20-22 kDa and 36 kDa). All 10 sera reacted to the protein at MW 20-22 kDa, 2 sera with MW 11-13 kDa, 3 sera with 16-17 kDa and 6 sera with 36 kDa. This study has thus demonstrated that spores of Bipolaris sp. contain allergenic components which may elicit IgE-mediated reactions.

House dust mite allergen levels in a Singapore hospital.

House dust mite allergens constitute one of the most important allergens in house dust. In this study, the levels of two common dust mite allergens, Der p I and Der f I, in a general hospital in Singapore were evaluated. Our results showed that these allergens were detected in 42/74 (or 57%) of the dust samples. Der p I was found to be the predominant allergen detected (p < 0.001). The allergen levels were, however, low with only 1/74 having a Der p I concentration above 2 micrograms g-1 dust. None of the samples had Der f I concentrations above this level. Of the various niches studied (mattresses, pillows, sofas, carpets, blinds and floors), the blinds and floors had the lowest concentration of allergen (p < 0.05). These low levels in the hospital compared to homes were attributed to the vigorous cleaning schedule in the hospital, the use of plastic to encased mattresses and pillows, vinyl covered sofas and vinyl lined floors. These practices may be adopted in the home as a means to reduce mite allergen exposure.

Field efficacy of mosquito coil formulations containing d-allethrin and d-transallethrin against indoor mosquitos especially Culex quinquefasciatus Say.

Comparative field efficacy studies of four mosquito coil formulations containing active ingredient of d-allethrin (0.19 or 0.28 w/w) and d-transallethrin (0.12 or 0.16% w/w) and blank coils without active ingredient were carried out in living rooms (mean size 54.1 m3) of residential houses in a squatter area in Butterworth, Malaysia. The major indoor biting mosquitos collected in the test site were that of Culex quinquefasciatus Say (84.7%). Mean percentage reduction of blank coils, coils with 0.19 and 0.28% d-allethrin and coils with 0.12 and 0.16% d-transallethrin were 29.0, 71.7, 70.9, 75.0 and 72.6%, respectively. The use of coils as a mean of personal protection against mosquitos is discussed.