Optimizing CRISPR/Cas9 for Gene Silencing of SOD1 in Mouse Models of ALS

Mutations in the SOD1 gene are the best characterized genetic cause of amyotrophic lateral sclerosis (ALS) and account for ~20% of inherited cases and 1-3% of sporadic cases. The gene-editing tool Cas9 can silence mutant genes that cause disease, but effective delivery of CRISPR-Cas9 to the central nervous system (CNS) remains challenging. Here, I developed strategies using canonical Streptococcus pyogenes Cas9 to silence SOD1. In the first strategy, I demonstrate effectiveness of systemic delivery of guide RNA targeting SOD1 to the CNS in a transgenic mouse model expressing human mutant SOD1 and Cas9. Silencing was observed in both the brain and the spinal cord. In the second strategy, I demonstrate the effectiveness of delivering both guide RNA and Cas9 via two AAVs into the ventricles of the brain of SOD1G93A mice. Silencing was observed in the brain and in motor neurons within the spinal cord. For both strategies, treated mice had prolonged survival when compared to controls. Treated mice also had improvements in grip strength and rotarod function. For ICV treated mice, we detected a benefit of SOD1 silencing using net axonal transport assays, a novel method to detect motor neuron function in mice before onset of motor symptoms. These studies demonstrate that Cas9-mediated genome editing can mediate disease gene silencing in motor neurons and warrants further development for use as a therapeutic intervention for SOD1-linked ALS patients

Miniature Multi-Sensor Array (mini-MSA) for Ground-to-Stratosphere Air Measurement, Phase II

There are numerous college research groups doing high altitude ballooning and working with drones. Many of these are making air quality measurements. Those measurements are effectively random. We are designing an inexpensive and easy to build assembly that others can build for under $500 with the goal of having a uniform data set that can be compared. This will provide more useful data for atmospheric studies. Our first version had too many noise issues and the gas sensor signals were unstable. In December 2020 Winsen Sensors (China) came out with a “multi-in-one sensor module,” the ZPHS01B. At a price point of approximately$150 we decided to completely revise our design

BMI Superior to Fat-Free Mass Index and Fat Mass Index for Predicting Arterial Stiffness in College Students

Arterial stiffness is a significant predictor of cardiovascular disease and mortality. Body mass index (BMI) has been used to predict cardiovascular disease outcomes but does not consider fat and lean mass. Fat mass index (FMI) and fat-free mass index (FFMI) may be superior to BMI in predicting arterial stiffness because it considers body composition. PURPOSE: This study aims to determine if FFMI and FMI are superior to BMI at predicting arterial stiffness in college students. METHODS: Participants came into the exercise physiology lab, signed consent, and then had anthropometric measurements of height and weight taken, as well as a body fat assessment (BOD POD, COSMED). Carotid-femoral pulse wave velocity (cfPWV), pulse wave analysis, and brachial and central blood pressure (BP) were then measured. Linear regression was used to determine if BMI, FFMI, and FMI predict BP and arterial stiffness. RESULTS: 237 college-aged (20.1 ± 3.3 yr.) participants (n = 122 males, n = 115 females) with an average BMI of 25.4 ± 4.1 kg/m2 completed this study. Results did not differ based on biological sex; thus, male and female data were aggregated. BMI significantly predicted brachial systolic BP (SBP) (r = .324), diastolic BP (DBP) (r = .223), central SBP (r =.374), DBP (r =.270), Augmentation pressure (AP) (r = .282), Augmentation index (AIX) (r = .213), and cfPWV (r = .258) (all P \u3c .05). FFMI significantly predicted brachial SBP (r = .395), central SBP (r = .320), and cfPWV (r = .224) (all P \u3c .05), but did not predict brachial or central DBP, AP or AIX. FMI significantly predicted central SBP (r = .143), brachial and central DBP (r =.329, r = .318, respectively), AP, (r = .275), and AIX (r = .328) (all P \u3c .05), but did not predict brachial SBP, or cfPWV. CONCLUSION: BMI appears to be a superior predictor of BP and arterial stiffness than FFMI and FMI in healthy college-aged adults

The relation between the diagonal entries and the eigenvalues of a symmetric matrix, based upon the sign pattern of its off-diagonal entries

It is known that majorization is a complete description of the relationships between the eigenvalues and diagonal entries of real symmetric matrices. However, for large subclasses of such matrices, the diagonal entries impose much greater restrictions on the eigenvalues. Motivated by previous results about Laplacian eigenvalues, we study here the additional restrictions that come from the off-diagonal sign-pattern classes of real symmetric matrices. Each class imposes additional restrictions. Several results are given for the all nonpositive and all nonnegative classes and for the third class that appears when n = 4. Complete description of the possible relationships are given in low dimensions. (C) 2012 Elsevier Inc. All rights reserved

Nonpositive Eigenvalues of the Adjacency Matrix and Lower Bounds for Laplacian Eigenvalues

Let $NPO(k)$ be the smallest number $n$ such that the adjacency matrix of any undirected graph with $n$ vertices or more has at least $k$ nonpositive eigenvalues. We show that $NPO(k)$ is well-defined and prove that the values of $NPO(k)$ for $k=1,2,3,4,5$ are $1,3,6,10,16$ respectively. In addition, we prove that for all $k \geq 5$, $R(k,k+1) \ge NPO(k) > T_k$, in which $R(k,k+1)$ is the Ramsey number for $k$ and $k+1$, and $T_k$ is the $k^{th}$ triangular number. This implies new lower bounds for eigenvalues of Laplacian matrices: the $k$-th largest eigenvalue is bounded from below by the $NPO(k)$-th largest degree, which generalizes some prior results.Comment: 23 pages, 12 figure

Locating the LCROSS Impact Craters

The Lunar CRater Observations and Sensing Satellite (LCROSS) mission impacted a spent Centaur rocket stage into a permanently shadowed region near the lunar south pole. The Sheperding Spacecraft (SSC) separated \sim9 hours before impact and performed a small braking maneuver in order to observe the Centaur impact plume, looking for evidence of water and other volatiles, before impacting itself. This paper describes the registration of imagery of the LCROSS impact region from the mid- and near-infrared cameras onboard the SSC, as well as from the Goldstone radar. We compare the Centaur impact features, positively identified in the first two, and with a consistent feature in the third, which are interpreted as a 20 m diameter crater surrounded by a 160 m diameter ejecta region. The images are registered to Lunar Reconnaisance Orbiter (LRO) topographical data which allows determination of the impact location. This location is compared with the impact location derived from ground-based tracking and propagation of the spacecraft's trajectory and with locations derived from two hybrid imagery/trajectory methods. The four methods give a weighted average Centaur impact location of -84.6796\circ, -48.7093\circ, with a 1{\sigma} un- certainty of 115 m along latitude, and 44 m along longitude, just 146 m from the target impact site. Meanwhile, the trajectory-derived SSC impact location is -84.719\circ, -49.61\circ, with a 1{\sigma} uncertainty of 3 m along the Earth vector and 75 m orthogonal to that, 766 m from the target location and 2.803 km south-west of the Centaur impact. We also detail the Centaur impact angle and SSC instrument pointing errors. Six high-level LCROSS mission requirements are shown to be met by wide margins. We hope that these results facilitate further analyses of the LCROSS experiment data and follow-up observations of the impact region.Comment: Accepted for publication in Space Science Review. 24 pages, 9 figure

VEGF promotes assembly of the p130Cas interactome to drive endothelial chemotactic signalling and angiogenesis

p130Cas is a polyvalent adapter protein essential for cardiovascular development, and with a key role in cell movement. In order to identify the pathways by which p130Cas exerts its biological functions in endothelial cells we mapped the p130Cas interactome and its dynamic changes in response to VEGF using high-resolution mass spectrometry and reconstruction of protein interaction (PPI) networks with the aid of multiple PPI databases. VEGF enriched the p130Cas interactome in proteins involved in actin cytoskeletal dynamics and cell movement, including actin-binding proteins, small GTPases and regulators or binders of GTPases. Detailed studies showed that p130Cas association of the GTPase-binding scaffold protein, IQGAP1, plays a key role in VEGF chemotactic signalling, endothelial polarisation, VEGF-induced cell migration, and endothelial tube formation. These findings indicate a cardinal role for assembly of the p130Cas interactome in mediating the cell migratory response to VEGF in angiogenesis, and provide a basis for further studies of p130Cas in cell movement