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Bioorthogonal click chemistry for fluorescence imaging of choline phospholipids in plants
Get PDFCitation: Paper, J. M., Mukherjee, T., & Schrick, K. (2018). Bioorthogonal click chemistry for fluorescence imaging of choline phospholipids in plants. Plant Methods, 14(1). https://doi.org/10.1186/s13007-018-0299-2Background: Phospholipids are important structural and signaling molecules in plant membranes. Some fluorescent dyes can stain general lipids of membranes, but labeling and visualization of specific lipid classes have yet to be developed for most components of the membrane. New techniques for visualizing membrane lipids are needed to further delineate their dynamic structural and signaling roles in plant cells. In this study we examined whether propargylcholine, a bioortholog of choline, can be used to label the major membrane lipid, phosphatidylcholine, and other choline phospholipids in plants. We established that propargylcholine is readily taken up by roots, and that its incorporation is not detrimental to plant growth. After plant tissue is harvested and fixed, a click-chemistry reaction covalently links the alkyne group of propargylcholine to a fluorescently-tagged azide, resulting in specific labeling of choline phospholipids. Results: Uptake of propargylcholine, followed by click chemistry with fluorescein or Alexa Fluor 594 azide was used to visualize choline phospholipids in cells of root, leaf, stem, silique and seed tissues from Arabidopsis thaliana. Colocalization with various subcellular markers indicated coinciding fluorescent signals in cell membranes, such as the tonoplast and the ER. Among different cell types in the leaf epidermis, guard cells displayed strong labeling. Mass spectrometry-based lipidomic analysis of the various plant tissues revealed that incorporation of propargylcholine was strongest in roots with approximately 50% of total choline phospholipids being labeled, but it was also incorporated in the other tissues including seeds. Phospholipid profiling confirmed that, in each tissue analyzed, incorporation of the bioortholog had little impact on the pool of choline plus choline-like phospholipids or other lipid species. Conclusion: We developed and validated a click-chemistry based method for fluorescence imaging of choline phospholipids using a bioortholog of choline, propargylcholine, in various cell-types and tissues from Arabidopsis. This click-chemistry method provides a direct way to metabolically tag and visualize specific lipid molecules in plant cells. This work paves the way for future studies addressing in situ localization of specific lipids in plants
Contribution of wetted clothing to body energy exchange and heat stress
Get PDFCitation: Elson, J., & Eckels, S. (2018). Contribution of wetted clothing to body energy exchange and heat stress. Journal of Thermal Biology, 78, 343–351. https://doi.org/10.1016/j.jtherbio.2018.09.014Quantifying the impact of clothing thermal and evaporation resistance is essential to providing representative boundary conditions for physiological modeling. In many models, sweat is assumed to drip off the skin surface to the environment and is not captured in clothing. In high metabolic rate and high temperature and humidity conditions the sweat produced by the body has the potential to saturate semipermeable clothing ensembles, changing the assumptions of the model. Workers, athletes and soldiers commonly wear encapsulating versions of such clothing to protect against environmental hazards. A saturated clothing model is proposed based on the ASHRAE two-node model using a saturated spot element in parallel with the existing method to account for sweat absorbed in the clothing. The work uses fundamental heat and mass transfer principles, modifying the existing formula using clothing measurements and basic assumptions. The effectiveness of the model is demonstrated by comparing the predictions of the original and proposed models, to the results of 21 soldiers exercising. The soldiers wore combat pants and shirt, helmet, gloves, shoes, socks, and underwear, and walked in a thermal chamber for 2 h at 42.2 °C dry bulb temperature, 54.4 °C wet bulb temperature, 20% relative humidity, and airspeed of 2 m/s. Core temperature, seven skin temperatures, heart rate, and total sweat loss were measured. The original model provides an average core temperature difference compared with the human subject results of 1.31 °C (SD = 0.557 °C) while the modified model improves the final prediction of core temperature to within an average of 0.15 °C (SD = 0.383 °C). The new model shows an improvement in the prediction of human core temperature under the tested conditions where dripping sweat will saturate clothing. The format can be used in multi-segmented thermal models and can continue to be developed and improved as more information on wetted clothing properties become available
Synthesis and Characterization of Multifunctional Branched Amphiphilic Peptide Bilayer Conjugated Gold Nanoparticles
Get PDFCitation: Natarajan, P., Sukthankar, P., Changstrom, J., Holland, C. S., Barry, S., Hunter, W. B., … Tomich, J. M. (2018). Synthesis and Characterization of Multifunctional Branched Amphiphilic Peptide Bilayer Conjugated Gold Nanoparticles. ACS Omega, 3(9), 11071–11083. https://doi.org/10.1021/acsomega.8b01633We provide strong chemical and biophysical evidence that documents that branched amphiphilic peptides, BAPs, known to assemble into spherical nanoassemblies in solution, do assemble as peptide-bilayer-delimited capsules. These nanoassemblies are termed branched amphiphilic peptide capsules (BAPCs). BAPCs are taken up by cells and accumulate in the perinuclear region to persist there without apparent degradation. BAPCs also entrap small proteins and solutes and stably encapsulate α-particle-emitting radionuclides. We have devised a method utilizing thiol chemistry to conjugate these peptide sequences onto gold nanoparticles (≤5 nm) with the objective of demonstrating the assembly of these peptides into a bilayer. The peptides are initially assembled as a monolayer on the gold surface via interaction with cysteine residues on the peptide C-terminus in an organic solvent. The subsequent transition of these peptide-monolayer-protected gold nanoparticles to an aqueous solution in the presence of excess peptides led to the formation of the peptide bilayer on the gold surface. The approach was exploited further to produce bilayer-coated magnetic nanoparticles. The innovation described in this study provides a stable metallic nanoparticle–peptide conjugate system that will help to determine interactions of BAPs in a biological system, with relative ease, important for developing future applications such as simultaneous delivery and imaging of surface-bound molecules of interest
Slowing-down and stopped charged particles cause angular dependence for absorbed dose measurements
Get PDFCitation: Bahadori, Amir A., Rajarshi Pal Chowdhury, Martin Kroupa, et al. (2018) Slowing-down and Stopped Charged Particles Cause Angular Dependence for Absorbed Dose Measurements. Radiation Physics and Chemistry. https://doi.org/10.1016/j.radphyschem.2018.06.012The space radiation environment is dominated by heavy charged particles with atomic numbers ranging from 1 to 93, with broad energy spectra that exceed 10 GeV per nucleon. Despite advances in space radiation modeling and transport, radiation detectors continue to provide critical data for understanding risks of health effects to astronauts in space. In the past, NASA relied on tissue-equivalent proportional counters and passive devices for operational dosimetry; however, in recent years, pixel detectors providing detailed information about the radiation environment through analysis of charged particle tracks have been demonstrated in space. These next-generation detectors, based on Timepix read-out technology, require special analysis considerations that were not necessary or possible for previous dosimetry tools. The impacts of slowing-down and stopped ions on absorbed dose measurements must be explicitly modeled to understand variations with detector orientation. The purpose of the present study is to conclusively demonstrate that while absorbed dose measurements of penetrating charged particles are independent of detector orientation, slowing-down and stopped particles can result in charged particle absorbed dose measurements that are dependent on detector orientation. Monte Carlo simulations of an unshielded detector, irradiated at selected orientations by different kinetic energy domains with fluence spectra representative of two historical solar particle events, are presented to demonstrate the dependence of absorbed dose measurements. Next, results from Monte Carlo simulations of the same energy domains and fluence spectra, isotropically impinging on an anisotropic shield configuration about the detector, are shown, to exhibit the potential for observing varying absorbed doses under realistic environment and shielding conditions. Finally, slowing-down and stopped proton data acquired with Timepix-based detectors at the Tandem Van de Graaff at Brookhaven National Laboratory are used to demonstrate the effect via accelerator-based measurements
Multi-dimensional Learning in the 5th Grade General Music Classroom: Supplemental Files
No full textMy lesson plan shown in my video teaching demonstration will show 5th graders performing a traditional song from the Apache tribe by singing, playing xylophones and other non-pitched percussion instruments, and playing a passing game. These students will accompany their voices by playing an ostinato pattern on the xylophone.
I have developed as a teacher through the Masters’ Program because I have learned new ways of teaching, especially when it comes to exposure to Orff and Kodaly methodologies, and improvisation within the classroom. I am now more comfortable incorporating improvisation, as well as music from many other cultures into my lessons, and my students are growing as musicians and as people due to it
Relativistic and resonant effects in the ionization of heavy atoms by ultra-intense hard X-rays
Get PDFCitation: Rudek, B., Toyota, K., Foucar, L., Erk, B., Boll, R., Bomme, C., … Rolles, D. (2018). Relativistic and resonant effects in the ionization of heavy atoms by ultra-intense hard X-rays. Nature Communications, 9(1), 4200. https://doi.org/10.1038/s41467-018-06745-6Availability of intense hard X-ray pulses allows exploration of multiple ionization effects in heavier elements. Here, the authors measure the complex charge state distributions of xenon and found a reasonable agreement by comparing with the model including the relativistic and resonance effects
Final Report
Get PDFThe Farmers Market and Local Foods Promotion Program (FMLFPP) funds projects across the United States, the U.S. territories, and the District of Columbia to increase access to and consumption of local foods. As part of this overarching goal, the program also supports training and technical assistance for direct-to-consumer outlets and local food businesses with the intent of developing or expanding market opportunities and intermediary functions such as aggregation, storage and distribution in the local food supply chain. Administered by U.S. Department of Agriculture (USDA) Agricultural Marketing Service (AMS), the program awarded 376 projects totaling more than $27 million in 2014.
AMS established a cooperative agreement with the Office of Educational Innovation and Evaluation at Kansas State University (KSU) to conduct a program evaluation of the 2014 grant projects. The evaluation had 3 primary objectives: 1) Describe successful outcomes and evidence supporting this attribution; 2) Illustrate the impact on the local food industry’s capacity regionally and nationally; and 3) Identify barriers that prevent the Program from addressing its primary purpose. The program evaluation incorporates quantitative and qualitative data gathered from the final performance reports (100 randomly selected reports manually reviewed, 355 reports5 reviewed by qualitative data software) and survey responses from 245 project managers (69% response rate). Final performance reports identified project impact immediately after grant project completion. A follow-up survey conducted by KSU in February, 2018, to project managers compiled projects’ long-term impacts.
This report provides comprehensive findings from the review of the 2014 projects’ Final performance reports as well as the survey responses. The findings summarize the impact of projects’ challenges, and lessons learned by program managers and AMS staff. Supporting data and documentation are available in the appendices as noted
Making an Inclusive Community Through Inclusive, Barrier-Free Theatre
Get PDFDrama therapists can work under many business structures when establishing a therapeutic theatre company to serve their clients and their communities. The business models the authors have found to provide homes for their acting companies are applicable to any type of therapeutic theatre. These include working under the auspices of already-established arts, educational, and service nonprofits to creating their own LLC or 501(c)3 nonprofit. This article offers action steps and advice so more drama therapists can establish their own theatre companies
Preliminary studies on bleaching of whey using ozone as a bleaching agent
Get PDFAnnatto is widely used in cheddar cheese manufacture to provide a consistent color and improve consumer preference. Only a portion of annatto added to cheese milk remains in cheese. The rest remains in the resultant liquid whey. Annatto in the liquid whey must be bleached in order to manufacture whey protein-based ingredients. Ozone, as a green bleaching source, has the potential to bleach annatto in liquid whey. The objective of this study was to examine the effectiveness of ozone in terms of holding time, temperature, and recirculation time as a whey bleaching agent. Annatto aqueous solution (0.017%) was prepared and incubated at 10, 20, 35 ℃. After that, the solution was exposed to ozone by pumping through an ozone generator in a continuous mode. The bleaching effect on annatto solution has an absorbance reduction of more than 95%. However, treatment temperature did not affect the bleaching efficiency in water solution. Sweet whey powder was reconstituted to 7.6% concentration and added with 0.017% annatto to simulate commercial cheese whey. Reconstituted liquid Whey was equilibrated to 10, 20, and 35 ℃ for 30 min and exposed to ozone by the ozone generator for 1 and 2 min in recirculation mode. In reconstituted cheese whey, when the circulation time was increased from 1 to 2 min, the absorbance reduction increased from 37% to 70%. On the other hand, when the temperature was increased from 10 to 20℃, the absorbance reduction reached a maximum of 80%. However, a further increase in temperature did not affect the annatto reduction. Overall, treatment temperature and recirculation time were found to be the most important factors that can affect bleaching of liquid whey using ozone
Comparison of Leaf Reflectance of Sorghum Plants Infested by Sugarcane Aphids
Get PDFSugarcane aphid, Melanaphis sacchari (Homiptera: Aphididae; Fig. 1A), is a significant pest to sorghum, Sorghum bicolor (Fig. 1B). Sorghum fields are regularly surveyed to detect pest infestation. Remote sensing methods using satellite and aircraft data are expected to provide an efficient alternative to field surveys. However, spectral signature of sugarcane aphid infested sorghum plants is not well understood. We evaluated reflectance spectra of sorghum leaves infested by sugarcane aphids. We used those spectral data to estimate indices that are related to photosynthetic pigments such as chlorophyll', anthocyanin, and carotenoids. We also estimated an aphid index, damage sensitive spectral index (DSSI)". We used these data to examine whether indices differ between pest infested and uninfested plants. We predicted that plants infested by aphid will have higher levels of anthocyanin, carotenoids, and DSSI indices compared to the control plants. In contrary, chlorophyll index will be greater for uninfested plants