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This document contains supplementary methods, tables, figures and references for the manuscript "ATP-competitive inhibitors midostaurin and avapritinib have distinct resistance profiles in exon 17-mutant KIT".

KIT is a type-3 receptor tyrosine kinase that is frequently mutated at exon 11 or 17 in a variety of cancers. First-generation KIT tyrosine kinase inhibitors (TKI) are ineffective against KIT exon 17 mutations, which favor an active conformation that prevents these TKIs from binding. The ATP-competitive inhibitors, midostaurin and avapritinib, which target the active kinase conformation, were developed to inhibit exon 17–mutant KIT. Because secondary kinase domain mutations are a common mechanism of TKI resistance and guide ensuing TKI design, we sought to define problematic KIT kinase domain mutations for these emerging therapeutics. Midostaurin and avapritinib displayed different vulnerabilities to secondary kinase domain substitutions, with the T670I gatekeeper mutation being selectively problematic for avapritinib. Although gatekeeper mutations often directly disrupt inhibitor binding, we provide evidence that T670I confers avapritinib resistance indirectly by inducing distant conformational changes in the phosphate-binding loop. These findings suggest combining midostaurin and avapritinib may forestall acquired resistance mediated by secondary kinase domain mutations.Significance:This study identifies potential problematic kinase domain mutations for next-generation KIT inhibitors midostaurin and avapritinib.

An obligatory step in the complex life cycle of the malaria parasite is sporogony, which occurs during the oocyst stage in adult female Anopheles mosquitoes. Sporogony is metabolically demanding, and successful oocyst maturation is dependent on host lipids. In insects, lipid energy reserves are mobilized by adipokinetic hormones (AKHs). We hypothesized that Plasmodium falciparum infection activates Anopheles gambiae AKH signaling and lipid mobilization. We profiled the expression patterns of AKH pathway genes and AgAkh1 peptide levels in An. gambiae during starvation, after blood feeding, and following infection and observed a significant time-dependent up-regulation of AKH pathway genes and peptide levels during infection. Depletion of AgAkh1 and AgAkhR by RNAi reduced salivary gland sporozoite production, while synthetic AgAkh1 peptide supplementation rescued sporozoite numbers. Inoculation of uninfected female mosquitoes with supernatant from P. falciparum-infected midguts activated AKH signaling. Clearly, identifying the parasite molecules mediating AKH signaling in P. falciparum sporogony is paramount.

KIT is a type-3 receptor tyrosine kinase that is frequently mutated at exon 11 or 17 in a variety of cancers. First-generation KIT tyrosine kinase inhibitors (TKI) are ineffective against KIT exon 17 mutations, which favor an active conformation that prevents these TKIs from binding. The ATP-competitive inhibitors, midostaurin and avapritinib, which target the active kinase conformation, were developed to inhibit exon 17-mutant KIT. Because secondary kinase domain mutations are a common mechanism of TKI resistance and guide ensuing TKI design, we sought to define problematic KIT kinase domain mutations for these emerging therapeutics. Midostaurin and avapritinib displayed different vulnerabilities to secondary kinase domain substitutions, with the T670I gatekeeper mutation being selectively problematic for avapritinib. Although gatekeeper mutations often directly disrupt inhibitor binding, we provide evidence that T670I confers avapritinib resistance indirectly by inducing distant conformational changes in the phosphate-binding loop. These findings suggest combining midostaurin and avapritinib may forestall acquired resistance mediated by secondary kinase domain mutations. SIGNIFICANCE: This study identifies potential problematic kinase domain mutations for next-generation KIT inhibitors midostaurin and avapritinib.

KIT is a type-3 receptor tyrosine kinase that is frequently mutated at exon 11 or 17 in a variety of cancers. First-generation KIT tyrosine kinase inhibitors (TKI) are ineffective against KIT exon 17 mutations, which favor an active conformation that prevents these TKIs from binding. The ATP-competitive inhibitors, midostaurin and avapritinib, which target the active kinase conformation, were developed to inhibit exon 17–mutant KIT. Because secondary kinase domain mutations are a common mechanism of TKI resistance and guide ensuing TKI design, we sought to define problematic KIT kinase domain mutations for these emerging therapeutics. Midostaurin and avapritinib displayed different vulnerabilities to secondary kinase domain substitutions, with the T670I gatekeeper mutation being selectively problematic for avapritinib. Although gatekeeper mutations often directly disrupt inhibitor binding, we provide evidence that T670I confers avapritinib resistance indirectly by inducing distant conformational changes in the phosphate-binding loop. These findings suggest combining midostaurin and avapritinib may forestall acquired resistance mediated by secondary kinase domain mutations. Significance: This study identifies potential problematic kinase domain mutations for next-generation KIT inhibitors midostaurin and avapritinib.

According to the Miami New Times Florida has the second fewest native residents of any state. Only 36 % of Florida’s population in 2012 are born Floridians. Newcomers to Florida often find it difficult to grow a garden in the sandy soil prevalent throughout the state. This presents an opportunity for Master Gardeners (MGs) to offer educational programs for residents to address identified needs. Among the objectives of this group are to teach residents how to build healthy soils and to explain their role in protecting the environment beginning with practices adopted in their backyard. This is achieved by using multiple venues, for instance, the extension office, garden clubs, homeowners’ associations, and public libraries, and presentation methods are utilized by MGs to teach youth and adult residents topics such as Building Healthy Soil, Composting, Vegetable Gardening, Lawn Care, and Pest Management. A year-end survey of residents participating in horticulture activities offered by the MGs showed 82 % (n-65) never took a soil test before program participation. This number declined to 25 % after the class. A total of 64 % of the respondents adopted to implement up to three gardening practices as a result of participating in horticulture programs offered by MGs and 14 % adopted four to six practices. Participation in educational activities offered by the MGs show an upward trend. For example, the Speakers’ Bureau has seen a 9 % increase in requests for educational talks between January and March 2015 compared to the same time frame in 2014. The emphasis placed on building healthy soil and the tools with which to do it are making the public more aware of the need to understand the environment as it relates to achieving productive vegetable gardens and a beautiful landscape.

Long-term monitoring of systems installed in the field is the ultimate standard for evaluating photovoltaic components and systems. This study, which involves the long-term outdoor exposure in a hot and humid climate, intends to address the performance degradation and failure mechanisms which are difficult or impossible to simulate in the lab during time constrained accelerated tests. Experimental data including irradiance, temperature, DC/AC current and voltage has been collected on diverse generations of photovoltaic modules installed throughout the state of Florida. Long term module reliability and lifetime are evaluated using a two pronged approach. 1) Modules have been deployed outdoors for long time periods with systematic - 15 minutes interval- climatic and performance measurements 2) Real-time climatic and performance measurements of modules following long-term outdoor exposed. Visual, IR and electrical insulation inspections were performed are also presented in this paper. Multiple analytical methods are used to quantify energy production and power degradation over time, including Performance Ratio analysis, and PVUSA regression analysis. Real-time field measurements were reviewed for both overall return rates and compare them with the nameplate performance values and to identify the failure mechanism that caused the return.

Extensive changes to mouthguards may occur as a result of wear and tear and equipment abuse.To examine how the shape and thickness of custom-fabricated mouthguards change over the course of a complete season of high school football.Repeated-measures, case-series study.High school.Fifteen high school varsity football players aged 15 to 17 years began the investigation; 4 participants lost their mouthguards before the end of the season.Custom-made mouthguards were examined before the start of the season, midway through the season, and at the end of the season to document changes to material thickness and shape. Along with evaluating the thickness of mouthguards at the incisor/canine and molar regions, we assessed shape retention by evaluating the width of the mouthguard channel at the first molar.Material thickness at the incisor/canine region decreased by approximately 16% (F(2,20) = 34.20, P.001) after 6 weeks of participation. Over the same time period, the mouthguard thickness at the molars decreased by 23% (F(2,20) = 31.92, P.001), and the shape of the mouthguards changed as well (F(1,11) = 6.01, P = .029).Although test mouthguards displayed significant deterioration in shape and material thickness, more research is needed to establish wearing length. Frequent inspections of mouthguards are advised to ensure that fit and protective abilities have not been compromised.

There is some concern regarding the amount of material thinning that occurs during the fabrication of custom-fabricated mouthguards. It is unclear if this thinning is merely a consequence of the fabrication process or related to other factors such as jaw size. Thus, the purpose of this study was to evaluate the contribution that various dimensional characteristics of the dental arch and the height of the stone model would have on mouthguard thinning. Fifteen subjects participated in this investigation. Alginate impressions from each subject were used to produce three replicas of the maxillary dentition with only the height of the base varying amongst them. The total height of the three models were 20, 25, and 30 mm. A single mouthguard was produced using each of the stone models. The material thickness of the mouthguard was assessed at the labial and occlusal surfaces. Additionally, the dimensions of the stone models were documented. Pearson product moment correlation coefficients were calculated to determine the linear relationship between material thickness and (i) the height of the stone models, (ii) the arch length and (iii) the area covered by the stone model. Statistical tests performed using the mean thickness values collected from the incisors and canines revealed a high negative correlation between the height of the stone model and material thickness (r = -0.82). In addition, a low to moderate positive linear correlation was noted between arch length and occlusal thickness at the molars (r = 0.57) and between the area of the stone model with the occlusal thickness (r = 0.49). The results of the present study indicate that the height of the model used to fabricate custom mouthguards should be kept as low as possible but still allow for the production of a properly fitting mouthguard.