A 680,000-person megastudy of nudges to encourage vaccination in pharmacies

Encouraging vaccination is a pressing policy problem. To assess whether text-based reminders can encourage pharmacy vaccination and what kinds of messages work best, we conducted a megastudy. We randomly assigned 689,693 Walmart pharmacy patients to receive one of 22 different text reminders using a variety of different behavioral science principles to nudge flu vaccination or to a business-as-usual control condition that received no messages. We found that the reminder texts that we tested increased pharmacy vaccination rates by an average of 2.0 percentage points, or 6.8%, over a 3-mo follow-up period. The most-effective messages reminded patients that a flu shot was waiting for them and delivered reminders on multiple days. The top-performing intervention included two texts delivered 3 d apart and communicated to patients that a vaccine was “waiting for you.” Neither experts nor lay people anticipated that this would be the best-performing treatment, underscoring the value of simultaneously testing many different nudges in a highly powered megastudy

The Somatic Genomic Landscape of Glioblastoma

We describe the landscape of somatic genomic alterations based on multi-dimensional and comprehensive characterization of more than 500 glioblastoma tumors (GBMs). We identify several novel mutated genes as well as complex rearrangements of signature receptors including EGFR and PDGFRA. TERT promoter mutations are shown to correlate with elevated mRNA expression, supporting a role in telomerase reactivation. Correlative analyses confirm that the survival advantage of the proneural subtype is conferred by the G-CIMP phenotype, and MGMT DNA methylation may be a predictive biomarker for treatment response only in classical subtype GBM. Integrative analysis of genomic and proteomic profiles challenges the notion of therapeutic inhibition of a pathway as an alternative to inhibition of the target itself. These data will facilitate the discovery of therapeutic and diagnostic target candidates, the validation of research and clinical observations and the generation of unanticipated hypotheses that can advance our molecular understanding of this lethal cancer

Reactivation of quiescent VZV in hESC-derived neurons induced by growth factor withdrawal.

<p>(A&C) hESC-derived neurons were incubated with low MOI VZV in the presence of ACV. Two weeks after exposure to virus and one week after removal of ACV, no GFP expression was detected in 50% of neuron-containing wells. At this time point, growth factors (GF) were withdrawn from the medium in wells that were GFP negative. By 4 days after GF withdrawal, massive loss of neurites was observed (B), eventually resulting in death of the cells in the wells by day 5 after treatment. However 30% of the initially GFP negative wells receiving GF-withdrawal treatment contained single and small foci of ORF66 protein-expressing neurons. (D). A and B are phase micrographs, C and D fluorescence micrographs of the same microscopic fields. Scale bar = 100μm.</p

RNA expression in productively and quiescently-infected hESC-derived neurons.

<p>A) The upper two histograms show the counts of reads from RNASeq analysis of quiescently-infected neurons and the lower two those of productively infected neurons aligned to the annotated vOKA genome. Note the difference in the Y-axis scale between the sets of histograms depicting productive and quiescently infected cultures. In the annotated genome at the bottom of the Fig, ORFs depicted in red (increased) and green (reduced) are those displaying statically significant differences in enrichment between quiescent and productively infected neurons. B) Fold-changes between the relative expression of transcripts of VZV ORFs between quiescent and productively infected neurons. The duplicated genes of the short repeats region of the genome are enriched in quiescently-infected neurons. ORFs for which significant differences were detected are denoted by asterisks.</p

Reactivation of VZV in quiescently infected hESC-derived neurons by PI3K inhibition at 34°C results in a productive, spreading infection.

<p>(A-F) A microscopic field showing neurons expressing GFP after reactivation at 34°C for 2 (A,B), 6 (C,D) and 14 (E,F) days after initiation of treatment. GFP expression is first observed in individual neurons, and spreads over time in the same initial foci of expression. (G-H) Another experiment showing the results of LY-induced reactivation at 34°C. The area depicted by the box in G is presented at higher magnification in H, showing the diffuse filling of neurons with GFP at 34<b>°</b>C. (I-N) In another reactivation experiment at 34°C, a focus of GFP expression (I&L) initially spreads over 3 days (J&M), but then contracts over a period of 4 days (K&N). Scale bars = 100μm.</p

DNA fluorescent <i>in situ</i> hybridization confirms the presence of VZV genomes in nuclei of hESC-derived neurons quiescently infected with VZV.

<p>Neurons were infected with VZV-ORF66-GFP and <i>in situ</i> hybridization performed on isolated nuclei as described in the methods. (A) shows FISH of nuclei from productively and (B) from quiescently infected neurons. Almost all quiescently, infected FISH+ neurons contained only one puncta in their nuclei. (C) FISH for VZV genomes in quiescently infected neurons receiving the PI3K inhibitor LY as a reactivation stimulus. After LY treatment there was a slight decrease in the percentage of FISH+ nuclei in the preparations, but 25% of labeled nuclei contained 2 or more puncta, suggesting additional sites of VZV genomes had appeared. See <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1004885#ppat.1004885.t001" target="_blank">Table 1</a> for quantification of the puncta. Scale Bar = 5 μm.</p

Percentage of neuronal nuclei containing fluorescent puncta following DNA FISH to detect VZV genomes.

<p>Percentage of neuronal nuclei containing fluorescent puncta following DNA FISH to detect VZV genomes.</p

Quiescent infection and reactivation of VZV in hESC-derived neurons without the use of ACV.

<p>hESC-derived neurons were infected via their axons in compartmentalized microfluidic chambers as detailed in the methods. A) Quantification of VZV genomes and transcripts from ORF63 and ORF31 were quantified by digital qPCR from nuclei acids extracted from the cell-body compartment two weeks after infection. B) & C) Reactivation of VZV in axonally-infected neurons. (B) Reactivation of VZV in hESC-derived neurons by LY 2 weeks after quiescent axonal infection. Neurons infected as in A were treated for 4 days with LY at 37°C and DNA and RNA extracted. qPCR revealed an increase in VZV genomes and transcripts from ORF62 and ORF31. No GFP-positive neurons were observed under these reactivation conditions. (C) Reactivation of neurons axonally-infected by VZV at 34°C. A photomicrograph of a microfluidic chamber where the cell body compartment (CB) containing the somata of axonally infected neurons was treated for 4 days with LY at 34 degrees is shown. A cluster of neurons (upper box) expressing ORF66GFP as a result of the treatment (higher magnification image in upper inset). The axons in the axonal compartment (Ax) are coated by the GFP-fluorescent debris (lower box, higher magnification image in the lower inset) used to infect the axons. Ch = microfluidic channels connecting the cell body and axonal compartments. Scale Bars = 100μm.</p

Reactivation stimuli increase the number of VZV genomes and transcripts in quiescently-infected human neurons.

<p>Wells of quiescently infected neurons were induced to reactivate VZV using growth factor withdrawal (GF n = 2 for each time point) or treatment with PI3K inhibitor LY294002 (LY) 2, 4, or 7 weeks (n = 5 for each time point) after infection. DNA and RNA were extracted from the wells and VZV genomes (A) and transcripts (B) of ORF63 and ORF31 were quantified. Both treatments increased the levels of both viral genomes and transcripts to varying degrees at all time points tested, indicating at least a partial reactivation of VZV. In all experiments reactivating VZV with LY, changes in nucleic acid levels measured were statistically significant.</p