Efficient generation of mNeonGreen <i>Plasmodium falciparum</i> reporter lines enables quantitative fitness analysis

CRISPR editing has enabled the rapid creation of fluorescent Plasmodium transgenic lines, facilitating a deeper understanding of parasite biology. The impact of genetic perturbations such as gene disruption or the introduction of drug resistance alleles on parasite fitness is typically quantified in competitive growth assays between the query line and a wild type reference. Although fluorescent reporter lines offer a facile and frequently used method to measure relative growth, this approach is limited by the strain background of the existing reporter, which may not match the growth characteristics of the query strains, particularly if these are slower-growing field isolates. Here, we demonstrate an efficient CRISPR-based approach to generate fluorescently labelled parasite lines using mNeonGreen derived from the LanYFP protein in Branchiostoma lanceolatum, which is one of the brightest monomeric green fluorescent proteins identified. Using a positive-selection approach by insertion of an in-frame blasticidin S deaminase marker, we generated a Dd2 reporter line expressing mNeonGreen under the control of the pfpare (P. falciparum Prodrug Activation and Resistance Esterase) locus. We selected the pfpare locus as an integration site because it is highly conserved across P. falciparum strains, expressed throughout the intraerythrocytic cycle, not essential, and offers the potential for negative selection to further enrich for integrants. The mNeonGreen@pare line demonstrates strong fluorescence with a negligible fitness defect. In addition, the construct developed can serve as a tool to fluorescently tag other P. falciparum strains for in vitro experimentation

THE DETERMINATION OF NOVEL IMPACT CONDITIONS FOR THE ASSESMENT OF LINEAR AND ANGULAR HEADFORM ACCELERATIONS

Sports helmets, albeit very effective at preventing traumatic brain injury, have not mitigated the risk of mild traumatic brain injury in sport (Flik, Lyman, & Marx, 2005). Current protocols utilized in sports helmet testing incorporate only impact vectors through the center of mass, eliciting primarily linear accelerations. Angular acceleration has been suggested to be a better predictor of diffuse head injury than linear acceleration (Holbourn, 1943); therefore, the objective of this study was to develop a protocol capable of producing and measuring both forms of acceleration for future implementation into sports helmet standards

THE EFFECT OF IMPACT CONDITION ON THE RELATIONSHIP BETWEEN LINEAR AND ANGULAR ACCELERATION LINEAR

Helmets are mandatory in many contact sports and are designed to prevent traumatic brain injuries. When assessing their performance, angular acceleration is not measured, as it is generally assumed to be highly correlated with linear acceleration (Pellman et al., 2003). Although being common, this assumption is not supported by strong data. The aim of this study was to establish the relationship between linear and angular acceleration

PERCEIVED DIFFERENCES IN SKATING CHARACTERISTICS RESULTING FROM THREE CROSS SECTIONAL SKATE BLADE PROFILES

The purpose of this study was to document differences in perceived skating characteristics resulting from three unique cross sectional skate blade profiles. Sixteen (n=16) University level hockey players were used in this double blind study looking at the perceived performance differences of four different skate blade profiles. No significant differences were found between skate blade profiles, preferred skate blade profile and time to complete given drills. Future research should look at different blade profiles and their interaction at ice level

Nonlocal calculation for nonstrange dibaryons and tribaryons

We study the possible existence of nonstrange dibaryons and tribaryons by solving the bound-state problem of the two- and three-body systems composed of nucleons and deltas. The two-body systems are $NN$, $N\Delta$, and $\Delta\Delta$, while the three-body systems are $NNN$, $NN\Delta$, $N\Delta\Delta$, and $\Delta\Delta\Delta$. We use as input the nonlocal $NN$, $N\Delta$, and $\Delta\Delta$ potentials derived from the chiral quark cluster model by means of the resonating group method. We compare with previous results obtained from the local version based on the Born-Oppenheimer approximation.Comment: 19 pages. To be published in Physical Review

Nuclear Force from Monte Carlo Simulations of Lattice Quantum Chromodynamics

The nuclear force acting between protons and neutrons is studied in the Monte Carlo simulations of the fundamental theory of the strong interaction, the quantum chromodynamics defined on the hypercubic space-time lattice. After a brief summary of the empirical nucleon-nucleon (NN) potentials which can fit the NN scattering experiments in high precision, we outline the basic formulation to derive the potential between the extended objects such as the nucleons composed of quarks. The equal-time Bethe-Salpeter amplitude is a key ingredient for defining the NN potential on the lattice. We show the results of the numerical simulations on a $32^4$ lattice with the lattice spacing $a \simeq 0.137$fm (lattice volume (4.4 fm)$^4$) in the quenched approximation. The calculation was carried out using the massively parallel computer Blue Gene/L at KEK. We found that the calculated NN potential at low energy has basic features expected from the empirical NN potentials; attraction at long and medium distances and the repulsive core at short distance. Various future directions along this line of research are also summarized.Comment: 13 pages, 4 figures, version accepted for publication in "Computational Science & Discovery" (IOP

Spin observables of the reactions NN -> DeltaN and pd -> Delta (pp)(1S0) in collinear kinematics

A general formalism for double and triple spin-correlations of the reaction NN -> DeltaN is developed for the case of collinear kinematics. A complete polarization experiment allowing to reconstruct all of the four amplitudes describing this process is suggested. Furthermore, the spin observables of the inelastic charge-exchange reaction pd -> Delta^0(pp)(1S0) are analyzed in collinear kinematics within the single pN scattering mechanism involving the subprocess pn -> Delta^0p. The full set of spin observables related to the polarization of one or two initial particles and one final particle is obtained in terms of three invariant amplitudes of the reaction pd -> Delta (pp)(1S0) and the transition form factor d->(pp)(1S0). A complete polarization experiment for the reaction pd -> Delta^0(pp)(1S0) is suggested which allows one to determine three independent combinations of the four amplitudes of the elementary subprocess NN -> DeltaN.Comment: 12 pages, 1 figur

Hadron interactions in lattice QCD

Progress on the potential method, recently proposed to investigate hadron interactions in lattice QCD, is reviewed. The strategy to extract the potential in lattice QCD is explained in detail. The method is applied to extract $NN$ potentials, hyperon potentials and the meson-baryon potentials. A theoretical investigation is made to understand the origin of the repulsive core using the operator product expansion. Some recent extensions of the method are also discussed.Comment: 52 pages, 65 figures, Review to appear in "Progress in Particle and Nuclear Physics

A comparison in a youth population between those with and without a history of concussion using biomechanical reconstruction

OBJECTIVE: Concussion is a common topic of research as a result of the short- and long-term effects it can have on the affected individual. Of particular interest is whether previous concussions can lead to a biomechanical susceptibility, or vulnerability, to incurring further head injuries, particularly for youth populations. The purpose of this research was to compare the impact biomechanics of a concussive event in terms of acceleration and brain strains of 2 groups of youths: those who had incurred a previous concussion and those who had not. It was hypothesized that the youths with a history of concussion would have lower-magnitude biomechanical impact measures than those who had never suffered a previous concussion. METHODS: Youths who had suffered a concussion were recruited from emergency departments across Canada. This pool of patients was then separated into 2 categories based on their history of concussion: those who had incurred 1 or more previous concussions, and those who had never suffered a concussion. The impact event that resulted in the brain injury was reconstructed biomechanically using computational, physical, and finite element modeling techniques. The output of the events was measured in biomechanical parameters such as energy, force, acceleration, and brain tissue strain to determine if those patients who had a previous concussion sustained a brain injury at lower magnitudes than those who had no previously reported concussion. RESULTS: The results demonstrated that there was no biomechanical variable that could distinguish between the concussion groups with a history of concussion versus no history of concussion. CONCLUSIONS: The results suggest that there is no measureable biomechanical vulnerability to head impact related to a history of concussions in this youth population. This may be a reflection of the long time between the previous concussion and the one reconstructed in the laboratory, where such a long period has been associated with recovery from injury