Pneumococcal epidemiology among us adults hospitalized for community-acquired pneumonia

BACKGROUND: Few studies have measured the burden of adult pneumococcal disease after the introduction of 13-valent pneumococcal conjugate vaccine (PCV13) into the US infant vaccination schedule. Further, most data regarding pneumococcal serotypes are derived from invasive pneumococcal disease (IPD), which represents only a fraction of all adult pneumococcal disease burden. Understanding which pneumococcal serotypes cause pneumonia in adults is critical for informing current immunization policy. The objective of this study was to measure the proportion of radiographically-confirmed (CXR+) community-acquired pneumonia (CAP) caused by PCV13 serotypes in hospitalized US adults. METHODS: This observational, prospective surveillance study recruited hospitalized adults aged \u3e /=18years from 21 acute care hospitals across 10 geographically-dispersed cities in the United States between October 2013 and September 2016. Clinical and demographic data were collected during hospitalization. Vital status was ascertained 30days after enrollment. Pneumococcal serotypes were detected via culture from the respiratory tract and normally-sterile sites (including blood and pleural fluid). Additionally, a novel, Luminex-based serotype-specific urinary antigen detection (UAD) assay was used to detect serotypes included in PCV13. RESULTS: Of 15,572 enrolled participants, 12,055 eligible patients with CXR+CAP were included in the final analysis population. Mean age was 64.1years and 52.7% were aged \u3e /=65years. Common comorbidities included chronic obstructive pulmonary disease (43.0%) and diabetes mellitus (28.6%). PCV13 serotypes were detected in 552/12,055 (4.6%) of all patients and 265/6347 (4.2%) of those aged \u3e /=65years. Among patients aged 18-64years PCV13 serotypes were detected in 3.8-5.3% of patients depending on their risk status. CONCLUSIONS: After implementation of a pneumococcal conjugate vaccination program in US children, and despite the herd protection observed in US adults, a persistent burden of PCV13-type CAP remains in this population

INVESTIGATING THE CLASSICAL NON-HOMOLOGOUS END JOINING PATHWAY IN THE VIRAL VECTOR AEDES AEGYPTI

Aedes aegypti is a critical viral vector and is known to transmit yellow fever, dengue and Zika viruses. Current control mechanisms rely heavily on source removal and insecticide use making novel genetic control mechanisms an attractive alternative. The CRISPR/Cas9 system enhances basic and applied research by allowing for sequence specific genetic modification. Utilization of the CRISPR/Cas9 system relies on repair of DNA double stranded breaks by endogenous DNA repair pathways such as the classical non-homologous end joining (C-NHEJ) or homology directed repair (HDR). Manipulating the C-NHEJ pathway to increase rates of HDR would expedite experiments which rely on site specific integration of genetic constructs. To investigate the C-NHEJ pathway we aimed to create a luminescence-based assay to quantify shifts in end joining based repair and knockout the C-NHEJ genes ku70 and lig4 in the Ae. aegypti LVP strain. Using a GFP-firefly luciferase fusion protein, we created a luminescence-based assay which quantifies shifts in error-prone end joining repair in both Ae. aegypti embryos and cell lines (Aag2 and A20). Knockout of ku70 resulted in female-specific pupal lethality, reduced longevity in both male and female adult populations and delayed chorion melanization. After maintaining mutants ku70-/- for 25 generations, no shift in either the single strand annealing pathway (SSA) or NHEJ was detected in ku70-/- embryos. While no shift in SSA or C-NHEJ was detected in ku70-/- embryos, ku70-/- larvae were significantly more sensitive DNA DSB generated by the DNA DSB inducing antibiotic bleomycin when compared to our LVP strain (p<0.01, compared to ku70-/- G0s (66% vs 4.1%). The results of knockout of lig4 and ku70 suggest that CNHEJ plays an important role in both fitness and DNA damage sensitivity. Additionally, creation of the NHEJ assay will allow for future high throughput analysis of additional genes involved in DNA DSB repair in both Ae. aegypti embryos and cells. This work will hopefully aid in future DNA damage repair studies which may be used to improve both basic and applied research in Ae. aegypti