Single-cell DNA methylation sequencing by combinatorial indexing and enzymatic DNA methylation conversion.

BACKGROUND: DNA methylation is a critical molecular mark involved in cellular differentiation and cell-specific processes. Single-cell whole genome DNA methylation profiling methods hold great potential to resolve the DNA methylation profiles of individual cell-types. Here we present a method that couples single-cell combinatorial indexing (sci) with enzymatic conversion (sciEM) of unmethylated cytosines. RESULTS: The sciEM method facilitates DNA methylation profiling of single-cells that is highly correlated with single-cell bisulfite-based workflows (r2 > 0.99) whilst improving sequencing alignment rates, reducing adapter contamination and over-estimation of DNA methylation levels (CpG and non-CpG). As proof-of-concept we perform sciEM analysis of the temporal lobe, motor cortex, hippocampus and cerebellum of the human brain to resolve single-cell DNA methylation of all major cell-types. CONCLUSION: To our knowledge sciEM represents the first non-bisulfite single-cell DNA methylation sequencing approach with single-base resolution.

Inhaler Competence in Nepalese Patients with Asthma and Chronic Obstructive Pulmunory Disease: A Systematic Review.

BACKGROUND: Incorrect use of inhalers in patients with asthma and chronic obstructive pulmonary disease is prevalent internationally. This review aims to determine the prevalence and associated factors of incorrect inhaler use and effectiveness of intervention to improve the correct use among Nepalese patients with asthma and chronic obstructive pulmonary disease. METHODS: The protocol was registered in PROSPERO. Systematic literature search was performed in PubMed, Embase, CINAHL,and Google Scholar using predefined search terms. Studies in patients with asthma or chronic obstructive pulmonary disease, reporting at least one overall or critical error, using dry powder inhalers and pressurized metered dose inhalers were included. The quality of included studies was assessed using the National Heart, Lung, and Blood Quality Assessment Tools. A descriptive narrative synthesis was undertaken. RESULTS: Twelve studies were eligible for analysis. At least one overall and critical step of the inhaler technique was performed incorrectly by the large number of patients, ranging from 64-100% and 9-100%, respectively. The incorrect use was associated with factors related to patients, inhalers, and health care professionals. The interventions included a combination of verbal instruction, physical demonstration, and/or face-to-face training on correct inhaler use. Following the intervention, an improvement of 23-37% in the overall inhaler technique and 7-42% in the critical steps was achieved. CONCLUSIONS: Existing literature suggests that Nepalese patients with asthma and chronic obstructive pulmonary disease have high overall and critical error rates in the use of dry powder inhalers and pressurized metered dose inhalers. A well-designed educational intervention is necessary to improve the correct use of inhalers in this population.

Predictors of Mortality in Scrub Typhus Infection Requiring Intensive Care Admission in Tertiary Healthcare Centre of Nepal.

INTRODUCTION: This study aimed to explore the predictors of mortality from scrub typhus infection in patients requiring intensive care unit (ICU) admission. MATERIALS AND METHODS: A retrospective study was conducted on 120 patients with serum ELISA IgM positive for scrub typhus (optical density ≥ 0.5) admitted at the medical ICU of Chitwan Medical College Teaching Hospital between April 2016 and September 2017. Data was extracted from patient medical records and electronic database of the hospital. The outcome measurement was mortality (Yes/No) due to the infection. A multivariate binary logistic regression analysis (p < 0.10) using potential variables from bivariate analysis (p < 0.25) was adjusted to predict the mortality. RESULTS: The mortality rate was 20% (24/120). Factors associated with mortality, as found using bivariate analysis, were heart rate > 100/minute (p < 0.001), systolic blood pressure < 90 mmHg (p = 0.025), diastolic blood pressure < 60 mmHg (p = 0.032), serum creatinine > 1.4 mg/dl (p < 0.001), acute kidney injury requiring dialysis (p = 0.029), acute respiratory distress syndrome (p < 0.001), and shock requiring vasopressor (p < 0.001). Regression analysis showed age (odds ratio [OR] = 1.063; 95% CI = 1.010-1.118; p = 0.019) and serum creatinine (OR = 1.063; 95% CI = 1.010-1.118; p = 0.019) as significant predictors of poor outcome. CONCLUSION: Older age and high serum creatinine were found to be independent predictors of poor outcome in patients with scrub typhus admitted in medical ICU.