Identification of the Tembusu Virus in Mosquitoes in Northern Thailand.

Among emerging zoonotic pathogens, mosquito-borne viruses (MBVs) circulate between vertebrate animals and mosquitoes and represent a serious threat to humans via spillover from enzootic cycles to the human community. Active surveillance of MBVs in their vectors is therefore essential to better understand and prevent spillover and emergence, especially at the human-animal interface. In this study, we assessed the presence of MBVs using molecular and phylogenetic methods in mosquitoes collected along an ecological gradient ranging from rural urbanized areas to highland forest areas in northern Thailand. We have detected the presence of insect specific flaviviruses in our samples, and the presence of the emerging zoonotic Tembusu virus (TMUV). Reported for the first time in 1955 in Malaysia, TMUV remained for a long time in the shadow of other flaviviruses such as dengue virus or the Japanese encephalitis virus. In this study, we identified two new TMUV strains belonging to cluster 3, which seems to be endemic in rural areas of Thailand and highlighted the genetic specificities of this Thai cluster. Our results show the active circulation of this emerging flavivirus in Thailand and the need for continuous investigation on this poorly known but threatening virus in Asia.

Effects of the Developing and Using a Model to Predict Dengue Risk Villages Based on Subdistrict Administrative Organization in Southern Thailand.

The purpose of this study was to evaluate the effects of developing and using a model to predict dengue risk in villages and of a larval indices surveillance system for 2372 households in 10 Thai villages. A community participatory action research method was used in five steps: (1) community preparation covering all stakeholders, (2) assessment of the understanding of a dengue solution and a larval indices surveillance system, (3) development of a prediction and intervention model for dengue risk villages, (4) implementation of the model that responds to all stakeholders, and (5) evaluation of the effects of using the model. The questionnaires to assess and evaluate were validated and reliability tested. The chi-square test and Fisher's exact test were used to analyze the quantitative data collected by means of questionnaires. Thematic analysis was applied to the qualitative data collected through interviews. The results found that the model consisted of six main activities, including (1) setting team leader responsibility, (2) situation assessment, (3) prediction of the dengue risk in villages, (4) the six steps of the larval indices surveillance system, (5) the understanding of the dengue solution and the understanding of the larval indices surveillance system training program, and (6) local wisdom innovation. The effects of using the model showed a statistically significant increase in correct understanding among 932 family leaders, 109 village health volunteers, and 59 student leaders regarding dengue prevention and control (p < 0.05). The larval indices and dengue morbidity were diminished and related to the nine themes present in the community leaders' reflections and to the satisfaction of the community members. Hence, local administrative organizations should use community-based approaches as the subdistrict dengue solution innovation to reduce the dengue problem.

Understanding dengue solution and larval indices surveillance system among village health volunteers in high- and low-risk dengue villages in southern Thailand.

Background: Successful dengue solutions require community collaboration between agencies engaged in human health, vector control and the environment. In Thailand, village health volunteers emphasize the need for a health working group to interact, collaborate, and coordinate actions. The objectives of this study were to acquire an understanding of dengue solutions, as well as the larval indices surveillance system of village health volunteers in high- and low-risk dengue villages. Methods: After 12 months of training in dengue prevention and setting larval indices surveillance systems, an analytical cross-sectional survey was conducted. A total of 117 villages were included in the 18 primary care facilities within one district in southern Thailand, and they were divided into 71 high-risk and 46 low-risk dengue villages. Sample size was determined using the G*power formula. The content validity index and reliability values of Cronbach's alpha coefficient for the questionnaires were 0.91 and 0.83, respectively. A random sampling approach was used to acquire data. The chi-square test, t-test, and odds ratio were used to assess the sample's level of understanding. Results: The study included 1302 village health volunteers, including 895 and 407 from high- and low-risk dengue communities, respectively. In total, 87.9% were female, 51.6% were 20-35 years old, 48.8% had worked as a village health volunteer for 11-20 years, 27.1% had an upper elementary education, and 59.1% had dengue in the previous 12 months. Understanding of the dengue solution and larval indices surveillance system varied across high- and low-risk dengue villages. Village health volunteers with a high level of understanding of the dengue solution and larval indies surveillance system were 1.064 and 1.504 times more likely to stay in high-risk dengue villages, respectively (odds ratio [OR] = 1.064, 95% confidence interval [CI]:0.798-1.419, p = 0.672 and OR = 1.504, 95% CI:1.044-2.167, p = 0.028). Conclusions: Village health volunteers require ongoing training to understand the prevention and control of dengue and larval indices surveillance systems, promote awareness, and monitor dengue in both high- and low-risk dengue villages.

New Insights into the Biology of the Emerging Tembusu Virus.

Reported for the first time in 1955 in Malaysia, Tembusu virus (TMUV) remained, for a long time, in the shadow of flaviviruses with human health importance such as dengue virus or Japanese encephalitis virus. However, since 2010 and the first large epidemic in duck farms in China, the threat of its emergence on a large scale in Asia or even its spillover into the human population is becoming more and more significant. This review aims to report current knowledge on TMUV from viral particle organization to the development of specific vaccines and therapeutics, with a particular focus on host-virus interactions.

Aberrant neutrophil function among heavy smokers and chronic obstructive pulmonary disease patients.

BACKGROUD: Chronic Obstructive Pulmonary Disease (COPD) is a chronic inflammation disease of the respiratory tract. The aberrant functions of neutrophils has been reported in COPD patients including respiratory burst and phagocytosis. Unfortunately, there is little evidence of neutrophil functions in healthy smokers who are considered a high risk group for COPD. OBJECTIVE: To determine the alteration of leukocytic parameters, intracellular reactive oxygen species (ROS) levels and phagocytosis percentages among smokers and COPD. METHOD: Sixteen smokers, 17 COPD patients and 10 healthy controls were recruited. The informed consent and recruitment process were ethically approved by the WU-IRB committee. Leukocyte percentages were analyzed by automated cell counter. Polymorphonuclear cells were purified by gradient centrifugation. The cell suspensions were incubated with FITC-conjugated bacteria to allow phagocytosis. Dichlorofluorescin diacetate was loaded to the cells followed by porbol-12-myristyl acetate stimulation to generate ROS. The percentages of phagocytosis and intracellular ROS levels were analyzed using flow cytometry. Mean of leukocyte numbers, mean fluorescence intensities and phagocytosis percentages were compared by t-test using SPSS (version 17). RESULTS: An increased ROS level of circulating neutrophils was observed among heavy smokers and COPD (p<0.05) compared to healthy controls. There was concordance to neutrophil percentages. However, the function on phagocytosis of neutrophils was not abolished in any group of smokers except COPD. CONCLUSIONS: Our findings suggested that the elevation of intracellular ROS generated by circulating neutrophils resulted in pools of inflammatory mediators among heavy smokers, which was suspected as one of the factors causing COPD in high risk people.

Differences in the constitutive and SIV infection induced expression of Siglecs by hematopoietic cells from non-human primates.

The expression of the Siglec family of molecules by hematopoietic cells from uninfected and SIV infected disease susceptible rhesus macaques (RM) and SIV infected disease resistant sooty mangabeys (SM) and for comparison humans was carried out. The predominant cell lineage in all three species expressing Siglec's was monocytes. The major finding by both a cross sectional and a prospective SIV infection study showed that, whereas monocytes from RM show marked increase in each Siglec constitutively expressed, monocytes from SM showed marked decreases in Siglec-1 expression. While monocytes from all three species constitutively expressed Siglec-3, human monocytes in addition expressed Siglec-5 and -9 and to a lower density 7, monocytes from RM expressed Siglec-7 and those from SM expressed Siglec-1. Monocytes from all three species, however, expressed mRNA for Siglec-1, -5, -7 and -9. The reasons for the failure to detect these molecules at the protein level and the mechanisms for such distinct effects of SIV infection on Siglec expression are discussed.

A novel enhanced green fluorescent protein (EGFP)-K562 flow cytometric method for measuring natural killer (NK) cell cytotoxic activity.

Enhanced green fluorescent protein (EGFP) was stably expressed in human erythroleukaemia K562 cells (EGFP-K562) and used as target cells for measurement of natural killer (NK) cell cytotoxicity by flow cytometry. The compromised EGFP-K562 target cells were stained with propidium iodide (PI) and showed dual (green-red) fluorescent. Although the kinetic study demonstrated that the optimal incubation time for the assay was 4 h, a 2-h incubation period also gave comparable results. This new technique correlated strongly with the standard chromium (51Cr) release assay at the correlation coefficients of 0.87 and 0.89 at p-value <0.001 for 2- and 4-h incubation times, respectively. The EGFP-K562 stable cell line provides a novel method to measure NK cytotoxicity by flow cytometry without pre-staining or pre-labeling target cells.