The Development of New Tools for the Investigation of Protein Function Using Photo-Reactive Unnatural Amino Acids

Reported here is the direct synthesis and application of unnatural amino acids for the development of exploratory tools for protein studies. This work takes advantage of an expanded genetic code to extract a more precise chemical understanding of protein function with novel additions to the unnatural amino acid catalogue, as well as the expansion of techniques with previously developed compounds. The photochemical crosslinker, [D11]-p-benzoylphenylalanine (pBpa), is synthesized for isotopic labeling in proteins. When [D11]-pBpa is co-incorporated into protein with [D0]-¬pBpa it is a mass spectral tool for rapid and conclusive identification of crosslinked fragments. Following enzymatic digestion the fingerprint of M, M+ 11 is readily identified allowing for rapid peak identification and the determined site of crosslink formation with single amino acid accuracy. In a means to extract a level of spatiotemporal control over fluorescent labeling of protein, the photo-protected unnatural amino acid, o-nitrobenzyl cysteine (ONBC), is introduced to a small amino acid tag sequence CCPGCC. This tag is required and specifically binds the pro-fluorescent compound 5-bis(1,3,2-dithiasolan-2-yl)fluorescein (FlAsH). This work takes advantage of the inability of FlAsH to bind the cysteine-tag motif in the presence of an ONBC mutation. The photo-protected amino acid is deprotected with light, affording natural cysteine and the successful binding of FlasH to the tetracysteine tag only following ultraviolet irradiation. Finally, fluorinated tyrosine derivatives are synthetically modified to contain photo-protecting groups, which act as a disguise during unnatural amino acid mutagenesis techniques. Fluorinated tyrosines are recognized by endogenous tyrosyl-tRNA synthetases and incorporated globally throughout a protein at tyrosine positions. To circumvent this problem the o-nitrobenzyl photo-protecting group is installed on the tyrosine derivatives 2-fluorotyrosine, 3-fluorotyrosine, and 2,6-difluorotyrosine. The directed evolution of an orthogonal amber-tRNA synthetase, specific for these unnatural amino acids, is performed, providing the translational machinery for site-specific incorporation of these compounds. Following expression of protein with the protected tyrosine derivatives, protein exposed to ultraviolet irradiation subsequently loses the protecting group affording the site-specific incorporation of fluorinated tyrosine. Fluorinated tyrosines are introduced to the critical trysoine residue in the chromophore of super-folder green fluorescent protein to determine how the altered pKa affects its fluorescent properties

Evaluating National Football League Draft Choices: The Passing Game

Recruiting competent personnel is crucial for the success of any organization, especially in competitive sports where the success of teams depends upon the quality of players selected. This paper examines whether football executives are able to forecast who will be the most successful quarterbacks and wide receivers. Our data base is constructed from the NFL drafts between 1974 and 2005. We use a variety of measures to determine the success of the players selected in those drafts. We conclude that, although their ability to rank the future performance of players is less than perfect, football executives are very successful in evaluating the talent of athletes. There was no evidence that teams that selected Awell@ also had competitive success.sports forecasts; personnel forecasts; National Football League; sports economics

Thermochemistry of Alane Complexes for Hydrogen Storage: A Theoretical and Experimental Investigation.

Knowledge of the relative stabilities of alane (AlH(3)) complexes with electron donors is essential for identifying hydrogen storage materials for vehicular applications that can be regenerated by off-board methods; however, almost no thermodynamic data are available to make this assessment. To fill this gap, we employed the G4(MP2) method to determine heats of formation, entropies, and Gibbs free energies of formation for 38 alane complexes with NH(3-n)R(n) (R = Me, Et; n = 0-3), pyridine, pyrazine, triethylenediamine (TEDA), quinuclidine, OH(2-n)R(n) (R = Me, Et; n = 0-2), dioxane, and tetrahydrofuran (THF). Monomer, bis, and selected dimer complex geometries were considered. Using these data, we computed the thermodynamics of the key formation and dehydrogenation reactions that would occur during hydrogen delivery and alane regeneration, from which trends in complex stability were identified. These predictions were tested by synthesizing six amine-alane complexes involving trimethylamine, triethylamine, dimethylethylamine, TEDA, quinuclidine, and hexamine and obtaining upper limits of ΔG° for their formation from metallic aluminum. Combining these computational and experimental results, we establish a criterion for complex stability relevant to hydrogen storage that can be used to assess potential ligands prior to attempting synthesis of the alane complex. On the basis of this, we conclude that only a subset of the tertiary amine complexes considered and none of the ether complexes can be successfully formed by direct reaction with aluminum and regenerated in an alane-based hydrogen storage system

Initial Hydraulic modelling and Levee Stability Analysis of the Triple M Ranch Restoration Project

“Advanced Watershed Science and Policy (ESSP 660)” is a graduate class taught in the Master of Science in Coastal and Watershed Science & Policy program at California State University Monterey Bay (CSUMB). In 2007, the class was taught in four 4-week modules, each focusing on a local watershed issue. This report is one outcome of one of those 4-week modules taught in the fall 2007 session. (Document contains 32 pages

Conversion of a 3D scene into video

Three-dimensional graphics capabilities of portable devices such as smartphones are limited by their hardware. Even smartphones that include graphics processing units (GPU) cannot produce the graphics processing performance of a device (e.g., PC) that uses a more powerful desktop-class graphics processor. This disclosure describes techniques to create 360° video from a specified three-dimensional scene on a virtual desktop computer using a web browser and advanced graphics hardware. The video generated using this technique can be stored on a video hosting website and is suitable to be rendered by devices that lack advanced graphics capabilities

Preseason Functional Test Scores are Associated with Future Sports Injury in Female Collegiate Athletes

Brumitt, J, Heiderscheit, B, Manske, R, Niemuth, PE, Mattocks, A, and Rauh, MJ. Preseason functional test scores are associated with future sports injury in female collegiate athletes. J Strength Cond Res 32(6): 1692–1701, 2018—Recent prospective cohort studies have reported preseason functional performance test (FPT) measures and associations with future risk of injury; however, the findings associated with these studies have been equivocal. The purpose of this study was to determine the ability of a battery of FPTs as a preseason screening tool to identify female Division III (D III) collegiate athletes who may be at risk for a noncontact time-loss injury to the lower quadrant (LQ = low back and lower extremities). One hundred six female D III athletes were recruited for this study. Athletes performed 3 FPTs: standing long jump (SLJ), single-leg hop (SLH) for distance, and the lower extremity functional test (LEFT). Time-loss sport-related injuries were tracked during the season. Thirty-two (24 initial and 8 subsequent) time-loss LQ injuries were sustained during the study. Ten of the 24 initial injuries occurred at the thigh and knee. At-risk athletes with suboptimal FPT measures (SLJ #79% ht; (B) SLH #64% ht; LEFT $118 seconds) had significantly greater rates of initial (7.2 per 1,000 athletic exposures [AEs]) and total (7.6 per 1,000 AEs) time-loss thigh or knee injuries than the referent group (0.9 per 1,000 AEs; 1.0 per 1,000 AEs, respectively). At-risk athletes were 9 times more likely to experience a thigh or knee injury (odds ratio [OR] = 9.7, confidence interval [CI]: 2.3–39.9; p = 0.002) than athletes in the referent group. At-risk athletes with a history of LQ sports injury and lower off-season training habits had an 18-fold increased risk of a time-loss thigh or knee injury during the season (adjusted OR = 18.7, CI: 3.0–118.1; p = 0.002). This battery of FPTs appears useful as a tool for identifying female D III athletes at risk of an LQ injury, especially to the thigh or knee region

Pengaruh Green Intellectual Capital Terhadap Business Sustainability

Economically, the company tries to make a profit as part of its natural motivation (business). Meanwhile, socially, the company must also have a beneficial impact on the community so that its existence gets social legitimacy. This study examines the effect of Green Intellectual Capital (GIC) on Business Sustainability (BS). In practice, green intellectual capital consists of Green Human Capital (GHC), Green Structural Capital (GSC), and Green Relational Capital (GRC) that reflects the intangible assets owned by the company including knowledge, wisdom, experience, and innovation with due regard to environmental welfare. This study uses 150 data samples of manufacturing companies in the period 2016-2018. Hypothesis testing uses SPSS statistical software version 24. The results of this study prove that only GHC and GRC have an influence on BS