Genome-Wide DNA Methylation Profiles Indicate CD8+ T Cell Hypermethylation in Multiple Sclerosis

Objective

Determine whether MS-specific DNA methylation profiles can be identified in whole blood or purified immune cells from untreated MS patients.

Methods

Whole blood, CD4+ and CD8+ T cell DNA from 16 female, treatment naïve MS patients and 14 matched controls was profiled using the HumanMethylation450K BeadChip. Genotype data were used to assess genetic homogeneity of our sample and to exclude potential SNP-induced DNA methylation measurement errors.

Results

As expected, significant differences between CD4+ T cells, CD8+ T cells and whole blood DNA methylation profiles were observed, regardless of disease status. Strong evidence for hypermethylation of CD8+ T cell, but not CD4+ T cell or whole blood DNA in MS patients compared to controls was observed. Genome-wide significant individual CpG-site DNA methylation differences were not identified. Furthermore, significant differences in gene DNA methylation of 148 established MS-associated risk genes were not observed.

Conclusion

While genome-wide significant DNA methylation differences were not detected for individual CpG-sites, strong evidence for DNA hypermethylation of CD8+ T cells for MS patients was observed, indicating a role for DNA methylation in MS. Further, our results suggest that large DNA methylation differences for CpG-sites tested here do not contribute to MS susceptibility. In particular, large DNA methylation differences for CpG-sites within 148 established MS candidate genes tested in our study cannot explain missing heritability. Larger studies of homogenous MS patients and matched controls are warranted to further elucidate the impact of CD8+ T cell and more subtle DNA methylation changes in MS development and pathogenesis.     

Presence of a Physician Safety Champion Is Associated with a Reduction in Urinary Catheter Utilization in the Pediatric Intensive Care Unit

Background

Safety champions are effective in a variety of safety initiatives; however, there are no reports of their role in hospital-acquired infections prevention.

Objective

We aimed to describe the association of the presence of a physician safety champion with our urinary catheter device utilization ratios (DUR) in the Pediatric Intensive Care Unit (PICU).

Methods

Our PICU has incidence rates of catheter-associated urinary tract infections (CAUTI) and urinary catheter DUR above the 90^th percentile. Using a quasi-experimental design, we compared our DUR when the PICU team was exposed and unexposed (champion’s maternity leaves) to a physician safety champion. Hospital acquired infection (HAI) surveillance of all PICU admissions between April 1^st 2009 and June 29^th 2013 was done prospectively. To ensure stable acuity of the patient population over time, we used the central venous catheter (CVC) DUR as a control.

Results

The urinary catheter DUR was 0.44 (95% confidence interval [CI] 0.42–0.45) during the unexposed period versus 0.39 (95%CI 0.38–0.40) during the exposed period, for an absolute difference of 0.05 (95%CI 0.03–0.06; p<0.0001). The overall CVC DUR increased from 0.57 (95%CI 0.55–0.58) during the unexposed period to 0.63 (95%CI 0.61–0.64) during the exposed period, an absolute increase of 0.06 (95%CI 0.04–0.08; p<0.0001). Comparing the exposed and unexposed periods, adjusting for time trend, we observed a 17% decrease in the urinary catheter DUR when the safety champion was present (odds ratio [OR] 0.83; 95%CI 0.77–0.90). The rate of catheter-associated urinary tract infections did not change.

Conclusions

The presence of a unit-based safety champion can have a positive impact on urinary catheter DUR in a PICU.     

Understanding Public Perceptions of the HPV Vaccination Based on Online Comments to Canadian News Articles

Background

Given the variation in human papillomavirus (HPV) vaccine coverage across Canada, and debate regarding delivery of HPV vaccines in Catholic schools, we studied online comments on Canadian news websites to understand public perceptions of HPV and HPV vaccine.

Methods

We searched English- and French-language Canadian news websites for 2012 articles that contained the terms “HPV” or “human papillomavirus.” Articles about HPV vaccinations that contained at least one comment were included. Two researchers independently coded comments, analyzing them for emerging themes.

Results

We identified 3073 comments from 1198 individuals in response to 71 news articles; 630 (52.6%) individuals expressed positive sentiments about HPV vaccination (2.5 comments/individual), 404 (33.7%) were negative (3.0 comments/individual), 34 (2.8%) were mixed (1.5 comments/individual) and 130 (10.8%) were neutral (1.6 comments/individual). Vaccine-supportive commenters believed the vaccine is safe and effective. Common themes in negative comments included concerns regarding HPV vaccine safety and efficacy, distrust of pharmaceutical companies and government, and belief that school-age children are too young for HPV vaccine. Many comments focused on whether the Catholic Church has the right to inform health policy for students, and discussion often evolved into debates regarding HPV and sexual behaviour. We noted that many individuals doubted the credibility of vaccine safety information.

Conclusion

The majority of commenters do not appear to be against HPV vaccination, but public health messaging that focuses on both the vaccine’s safety profile, and its use as a means to prevent cancer rather than sexually transmitted HPV infection may facilitate its acceptance.     

A Bacterial Acetyltransferase Destroys Plant Microtubule Networks and Blocks Secretion

The eukaryotic cytoskeleton is essential for structural support and intracellular transport, and is therefore a common target of animal pathogens. However, no phytopathogenic effector has yet been demonstrated to specifically target the plant cytoskeleton. Here we show that the Pseudomonas syringae type III secreted effector HopZ1a interacts with tubulin and polymerized microtubules. We demonstrate that HopZ1a is an acetyltransferase activated by the eukaryotic co-factor phytic acid. Activated HopZ1a acetylates itself and tubulin. The conserved autoacetylation site of the YopJ / HopZ superfamily, K289, plays a critical role in both the avirulence and virulence function of HopZ1a. Furthermore, HopZ1a requires its acetyltransferase activity to cause a dramatic decrease in Arabidopsis thaliana microtubule networks, disrupt the plant secretory pathway and suppress cell wall-mediated defense. Together, this study supports the hypothesis that HopZ1a promotes virulence through cytoskeletal and secretory disruption.     

Pyramid Screening: Combining Three Genetic Screens into One Efficient Screen for Shoot Regeneration Mutants in Arabidopsis thaliana

Gain-of-function genetic mutants are typically found by creating a non-permissive condition and screening for plants that overcome the stress. Separate genetic screens are conducted for each condition, a potentially time-consuming effort. In severed Arabidopsis thaliana leaves, high light, suboptimal hormone exposure and old age were each independently found to reduce frequency of shoot regeneration. Rather than conducting three separate mutant screens to dissect these three pathways, a laborious process, we hypothesized that we could undertake a single economical screen to retrieve mutations specific for each trait as well as cross-talk alleles between pathways. Instead of creating non-permissive conditions for each of our three traits of interest, we combined the three suboptimal stress conditions such that only when combined was shoot regeneration abolished. No one stress was primarily responsible for loss of our trait, thus ensuring that we could recover mutant alleles in any of the three pathways of interest. Screening of 18,000 mutagenized plants resulted in 12 SHOOTING UP (stu) mutants. Secondary screening revealed that we had recovered alleles that were both specific for a pathway (light, hormones or age) and which acted through multiple pathways. Our approach, which we refer to as pyramid screening, represents an economical method for mutant screening of multiple pathways in parallel (three screens in one) and has the potential to recover alleles that cross-talk between multiple pathways that underlie a complex trait such as organ regeneration. Pyramid screening should be widely applicable across species.