The Development of Tip Elimination and Its Effects on the Industry

Presentation covers a brief history of tipping, current status of tipping, forces for and against the elimination of tip credit, and future implications

An Update on the Current Status of Restaurant Tipping in the US

The tipped wage continues to be a point of contention among the various stakeholders in this form of compensation. A variety of views exist among consumers, tipped wage earners and operators. Social activists, the federal and local governments and industry associations have all made their positions known. Recent environmental circumstances have exacerbated this situation. This poster provides an update on the status of tipping in U.S. restaurants and illustrates the combined effect of these forces

Novel ATP-Independent RNA Annealing Activity of the Dengue Virus NS3 Helicase

The flavivirus nonstructural protein 3 (NS3) bears multiple enzymatic activities and represents an attractive target for antiviral intervention. NS3 contains the viral serine protease at the N-terminus and ATPase, RTPase, and helicase activities at the C-terminus. These activities are essential for viral replication; however, the biological role of RNA remodeling by NS3 helicase during the viral life cycle is still unclear. Secondary and tertiary RNA structures present in the viral genome are crucial for viral replication. Here, we used the NS3 protein from dengue virus to investigate functions of NS3 associated to changes in RNA structures. Using different NS3 variants, we characterized a domain spanning residues 171 to 618 that displays ATPase and RNA unwinding activities similar to those observed for the full-length protein. Interestingly, we found that, besides the RNA unwinding activity, dengue virus NS3 greatly accelerates annealing of complementary RNA strands with viral or non-viral sequences. This new activity was found to be ATP-independent. It was determined that a mutated NS3 lacking ATPase activity retained full-RNA annealing activity. Using an ATP regeneration system and different ATP concentrations, we observed that NS3 establishes an ATP-dependent steady state between RNA unwinding and annealing, allowing modulation of the two opposing activities of this enzyme through ATP concentration. In addition, we observed that NS3 enhanced RNA-RNA interactions between molecules representing the ends of the viral genome that are known to be necessary for viral RNA synthesis. We propose that, according to the ATP availability, NS3 could function regulating the folding or unfolding of viral RNA structures

Immune stealth-driven O2 serotype prevalence and potential for therapeutic antibodies against multidrug resistant Klebsiella pneumoniae

Emerging multidrug-resistant bacteria are a challenge for modern medicine, but how these pathogens are so successful is not fully understood. Robust antibacterial vaccines have prevented and reduced resistance suggesting a pivotal role for immunity in deterring antibiotic resistance. Here, we show the increased prevalence of Klebsiella pneumoniae lipopolysaccharide O2 serotype strains in all major drug resistance groups correlating with a paucity of anti-O2 antibodies in human B cell repertoires. We identify human monoclonal antibodies to O-antigens that are highly protective in mouse models of infection, even against heavily encapsulated strains. These antibodies, including a rare anti-O2 specific antibody, synergistically protect against drug-resistant strains in adjunctive therapy with meropenem, a standard-of-care antibiotic, confirming the importance of immune assistance in antibiotic therapy. These findings support an antibody-based immunotherapeutic strategy even for highly resistant K. pneumoniae infections, and underscore the effect humoral immunity has on evolving drug resistance

An efficient preparation of 1,2-dihydropyridazines through a Diels-Alder/palladium-catalysed elimination sequence

© 2019 Elsevier Ltd A convenient, scalable synthesis of 1,2-dihydropyridazines is presented, based on the Diels-Alder cycloaddition of 1-acetoxy-1,3-butadiene with a variety of azo compounds, followed by a palladium-catalysed elimination. The products are produced on multigram scale and the new method is particularly efficient and atom-economical when compared with previous preparations of 1,2-dihydropyridazines

An Update on the Current Status of Restaurant Tipping in the US

The tipped wage continues to be a point of contention among the various stakeholders in this form of compensation. A variety of views exist among consumers, tipped wage earners and operators. Social activists, the federal and local governments and industry associations have all made their positions known. Recent environmental circumstances have exacerbated this situation. This poster provides an update on the status of tipping in U.S. restaurants and illustrates the combined effect of these forces

Small field in-air output factors: the role of miniphantom design and dosimeter type

The commissioning of treatment planning systems and beam modeling requires measured input parameters. The measurement of relative output in-air, Sc is particularly difficult for small fields. The purpose of this study was to investigate the influence of miniphantom design and detector selection on measured Sc values for small fields and to validate the measurements against Monte Carlo simulations. Measurements were performed using brass caps (with sidewalls) or tops (no sidewalls) of varying heights and widths. The performance of two unshielded diodes (60012 and SFD), EBT2 radiochromic film, and a fiber optic dosimeter (FOD) were compared for fields defined by MLCs (5-100 mm) and SRS cones (4-30 mm) on a Varian Novalis linear accelerator. Monte Carlo simulations were performed to theoretically predict Sc as measured by the FOD. For all detectors, Sc agreed to within 1% for fields larger than 10 mm and to within 2.3% for smaller fields. Monte Carlo simulation matched the FOD measurements for all size of cone defined fields to within 0.5%. Miniphantom design is the most important variable for reproducible and accurate measurements of the in-air output ratio, Sc, in small photon fields (less than 30 mm). Sidewalls are not required for fields less than or equal too 30 mm and tops are therefore preferred over the larger caps. Unlike output measurements in water, Scp, the selection of detector type for Sc is not critical, provided the active dosimeter volume is small relative to the field siz

Small field diode correction factors derived using an air core fibre optic scintillation dosimeter and EBT2 film

There is no commercially available real-time dosimeter that can accurately measure output factors for field sizes down to 4 mm without the use of correction factors. Silicon diode detectors are commonly used but are not dosimetrically water equivalent, resulting in energy dependence and fluence perturbation. In contrast, plastic scintillators are nearly dosimetrically water equivalent. A fibre optic dosimeter (FOD) with a 0.8 mm3 plastic scintillator coupled to an air core light guide was used to measure the output factors for Novalis/BrainLab stereotactic cones of diameter 4-30 mm and Novalis MLC fields of width 5-100 mm. The FOD data matched the output factors measured by a 0.125 cm3 Semiflex ion chamber for the MLC fields above 30 mm and those measured with the EBT2 radiochromic film for the cones and MLC fields below 30 mm. Relative detector readings were obtained with four diode types (IBA SFD, EFD, PFD, PTW 60012) for the same fields. Empirical diode correction factors were determined by taking the ratio of FOD output factors to diode relative detector readings. The diodes were found to over-respond by 3%-16% for the smallest field. There was good agreement between different diodes of the same model number

Characterization of mAbs against Klebsiella pneumoniae type 3 fimbriae isolated in a target-independent phage display campaign

ABSTRACT We used phage display, antibody engineering and high-throughput assays, to identify antibody-accessible targets of Klebsiella pneumoniae. We report the discovery of mAbs binding to type 3 fimbrial proteins, including MrkA. We found that anti-MrkA mAbs were cross-reactive to a diverse panel of K. pneumoniae clinical isolates, representing different O-serotypes. mAbs binding to MrkA have previously been described and have been shown to provide prophylactic protection although only modest protection when dosed therapeutically in vivo in a murine lung-infection model. Here, we used a combination of binding and opsonophagocytic killing studies using a high-content imaging platform to provide a possible explanation for the modest therapeutic efficacy in vivo reported in that model. Our work shows that expression of K. pneumoniae type 3 fimbriae in in vitro culture is not homogenous within a bacterial population. Instead, sub-populations of bacteria that do, and do not, express type 3 fimbriae exist. In a high-content opsonophagocytic killing assay, we showed that MrkA targeting antibodies initially promote killing by macrophages, however over time this effect is diminished. We hypothesise the reason for this is that bacteria not expressing MrkA can evade opsonophagocytosis. Our data support the fact that MrkA is a conserved, immunodominant protein that is antibody accessible on the surface of K. pneumoniae and suggest that additional studies should evaluate the potential of using anti-MrkA antibodies in different stages of K. pneumoniae infection (different sites in the body) as well as against K. pneumoniae biofilms in the body during infection and associated with medical devices.This work was funded by AstraZeneca and the Cambridge Biomedical Research Centre. SKB was funded by a Cambridge AstraZeneca PhD studentship. The authors are grateful for the assistance of AstraZeneca scientists John Ferguson and Augustin Harvey for the preparation of monocyte-derived macrophages. The authors thank University of Cambridge scientist Sally Forrest for her assistance with high-content imaging

RNA Interference-Mediated Silencing of Mitotic Kinesin KIF14 Disrupts Cell Cycle Progression and Induces Cytokinesis Failure

KIF14 is a microtubule motor protein whose elevated expression is associated with poor-prognosis breast cancer. Here we demonstrate KIF14 accumulation in mitotic cells, where it associated with developing spindle poles and spindle microtubules. Cells at later stages of mitosis were characterized by the concentration of KIF14 at the midbody. Time-lapse microscopy revealed that strong RNA interference (RNAi)-mediated silencing of KIF14 induced cytokinesis failure, causing several rounds of endoreduplication and resulting in multinucleated cells. Additionally, less efficacious KIF14-specific short interfering RNAs (siRNAs) induced multiple phenotypes, all of which resulted in acute apoptosis. Our data demonstrate the ability of siRNA-mediated silencing to generate epiallelic hypomorphs associated with KIF14 depletion. Furthermore, the link we observed between siRNA efficacy and phenotypic outcome indicates that distinct stages during cell cycle progression are disrupted by the differential modulation of KIF14 expression