Establishing protocol for detecting methylation of Ubiquitin carboxyl terminal hydrolase 1 gene’s promoter in nasopharyngeal carcinoma

Background: The methylation of Ubiquitin Carboxyl Terminal Hydrolase 1 (UCHL1) gene’s promoter has been reported as the etiological factor of nasopharyngeal tumorigenesis. Purpose: This study is designed to establish a protocol for detecting methylation of UCHL1 gene’s promoter in nasopharyngeal carcinoma (NPC) in a Vietnamese population. Materials and methods: 10 samples of NPC biopsy tissues and 10 samples of non-cancerous swabs were collected from the local hospital. Chloroform/Phenol method and Nested-MSP assays were established to detect methylation of a target gene. Results: The isolated DNA reached purity and high concentration which were confirmed by the method of absorbance measurement at 260nm and 280nm. Additionally, the Nested-MSP products of methylation or unmethylation were analyzed and visualized in the agarose gel with the band of 169bps and 210bps, respectively. By sequencing, it was confirmed that the two sets of primer could distinguish the status of methylation and unmethylation of UCHL1 gene’s promoter. Conclusion: Our data suggested that the current protocol could successfully identify the status of methylation and/or unmethylation of UCHL1 gene’s promoter

Non-annual seasonality of influenza-like illness in a tropical urban setting.

BACKGROUND In temperate and sub-tropical climates, respiratory diseases exhibit seasonal peaks in winter. In the tropics, with no winter, peak timings are irregular. METHODS To obtain a detailed picture of influenza-like illness (ILI) patterns in the tropics, we established an mHealth study in community clinics in Ho Chi Minh City (HCMC). During 2009-2015, clinics reported daily case numbers via SMS, with a subset performing molecular diagnostics for influenza virus. This real-time epidemiology network absorbs 6,000 ILI reports annually, one or two orders of magnitude more than typical surveillance systems. A real-time online ILI indicator was developed to inform clinicians of the daily ILI activity in HCMC. RESULTS From August 2009 to December 2015, 63 clinics were enrolled and 37,676 SMS reports were received, covering approximately 1.8M outpatient visits. Approximately 10.6% of outpatients met the ILI case definition. ILI activity in HCMC exhibited strong non-annual dynamics with a dominant periodicity of 206 days. This was confirmed by time-series decomposition, step-wise regression, and a forecasting exercise showing that median forecasting errors are 30%-40% lower when using a 206-day cycle. In ILI patients from whom naso-pharyngeal swabs were taken, 31.2% were positive for influenza. There was no correlation between the ILI time series and the time series of influenza, influenza A, or influenza B (all p > 0.15). CONCLUSION This suggests, for the first-time, that a non-annual cycle may be an essential driver of respiratory disease dynamics in the tropics. An immunological interference hypothesis is discussed as a potential underlying mechanism. This article is protected by copyright. All rights reserved