Insight into the Brain-Specific Alpha Isoform of the Scaffold Protein SH2B1 and its Rare Obesity-Associated Variants

Obesity poses a major health problem since it increases the risk for type 2 diabetes, metabolic syndrome, heart disease, and cancer. Mutations in SH2B1 have been identified in patients exhibiting severe childhood obesity and insulin resistance. Mice deficient in SH2B1 exhibit a similar phenotype to the patients, suggesting an important role for SH2B1 in regulating energy homeostasis. SH2B1 is alternatively spliced, leading to four isoforms (alpha, beta, gamma, and delta) that share an N-terminal 631 amino acids but have unique C-terminal tails. The findings that all SH2B1 isoforms are expressed in the brain and neuronal expression of SH2B1 beta rescues the obese phenotype in SH2B1-deficient mice suggest that SH2B1 plays an important function in neurons. In further support of a neuronal function for SH2B1, SH2B1 enhances neurotrophic factor-induced neurite outgrowth and the human obesity-associated mutations impair the ability of SH2B1 to promote neurite outgrowth. To gain insight into critical functions of SH2B1, its isoforms, and the human obesity-associated mutations, the work in this thesis characterizes the ability of rare SH2B1 mutations associated with human obesity both shared by all isoforms and those specific to SH2B1 alpha to affect cellular actions of SH2B1. It further examines how the unique C-terminal tails of SH2B1 regulate the actions of the shared 631 N-terminal amino acids of SH2B1. Our collaborator I. S. Farooqi identified variants in SH2B1 encoding for R227C, G238C, R270W, E299G, or T546A and the SH2B1 alpha-specific mutations A663V, V695M, or A723V in individuals with severe-early onset obesity and insulin resistance. We determined that like SH2B1 beta, SH2B1 alpha enhanced growth hormone-stimulated motility of macrophages and the mutations T546A, A663V and A723V impaired that enhancement. Unlike SH2B1 beta, SH2B1 alpha was unable to enhance NGF-mediated neurite outgrowth and the mutations in SH2B1 alpha had no impact. However, mutations G238C, R270W, E299G and T546A impaired SH2B1 beta-enhancement of NGF-induced neurite outgrowth. This led us to conclude that variants can disrupt specific isoform function, suggesting novel regulatory roles in SH2B1 isoform function. The discrepancy between SH2B1 alpha and beta regulation of neurite outgrowth directed us to ask how the unique C-terminal tails of the alpha and beta isoforms affect the ability of SH2B1 to regulate NGF-induced neurite outgrowth. By comparing the actions of SH2B1 alpha and beta to those of the N-terminal 631 amino acids shared by all isoforms of SH2B1, we found that the alpha tail prevents: 1) the ability of SH2B1 to cycle through the nucleus, and 2) SH2B1 enhancement of NGF-induced neurite outgrowth, gene expression, and phosphorylation of NGF-receptor, TrkA, and downstream signaling proteins Akt and PLC gamma. These functions were restored when Tyr753 in the alpha tail was mutated to Phe, suggesting that phosphorylation of Tyr753 regulates SH2B1 alpha function. We provide evidence that TrkA phosphorylates Tyr55 and Tyr439 in both SH2B1 alpha and SH2B1 beta, and Tyr753 in SH2B1 alpha. Finally, co-expression of SH2B1 alpha inhibits the ability of SH2B1 beta to enhance NGF-induced neurite outgrowth. These results suggest that the C-terminal tails of SH2B1 isoforms are key determinants of their cellular roles and are regulated by phosphorylation. By providing valuable information about how seemingly minor differences between isoforms can have a profound impact on the function of that protein, my studies also provide important insight into the impact of differential splicing on neuron function.PHDCellular & Molecular BiologyUniversity of Michigan, Horace H. Rackham School of Graduate Studieshttps://deepblue.lib.umich.edu/bitstream/2027.42/137175/1/raymjoe_1.pd

Adjoint-based airfoil shape optimization in transonic flow

The primary focus of this work is efficient aerodynamic shape optimization in transonic flow. Adjoint-based optimization techniques are employed on airfoil sections and evaluated in terms of computational accuracy as well as efficiency. This study examines two test cases proposed by the AIAA Aerodynamic Design Optimization Discussion Group. The first is a two-dimensional, transonic, inviscid, non-lifting optimization of a Modified-NACA 0012 airfoil. The second is a two-dimensional, transonic, viscous optimization problem using a RAE 2822 airfoil. The FUN3D CFD code of NASA Langley Research Center is used as the ow solver for the gradient-based optimization cases. Two shape parameterization techniques are employed to study their effect and the number of design variables on the final optimized shape: Multidisciplinary Aerodynamic-Structural Shape Optimization Using Deformation (MASSOUD) and the BandAids free-form deformation technique. For the two airfoil cases, angle of attack is treated as a global design variable. The thickness and camber distributions are the local design variables for MASSOUD, and selected airfoil surface grid points are the local design variables for BandAids. Using the MASSOUD technique, a drag reduction of 72.14% is achieved for the NACA 0012 case, reducing the total number of drag counts from 473.91 to 130.59. Employing the BandAids technique yields a 78.67% drag reduction, from 473.91 to 99.98. The RAE 2822 case exhibited a drag reduction from 217.79 to 132.79 counts, a 39.05% decrease using BandAids. --Abstract, page iii

An Architecture for Scaling NVO Services to TeraGrid

The term "cyberinfrastructure" has been adopted by the US National Science Foundation to mean "advanced computing engines, data archives and digital libraries, observation and sensor systems, and other research and education instrumentation [linked] into a common framework". One of the largest awards in this program is the TeraGrid, a linkage of large supercomputer centers based on the Globus software. Another cyberinfrastructure program is the National Virtual Observatory, a linkage of astronomical data publishers into a service-oriented framework. There are different philosophies behind the TeraGrid and the NVO architecture. This note explains a proposed service-oriented architecture for TeraGrid nodes that is an attempt to bridge these ways of working, and a prototype instantiation at Caltech

Pricing corn silage

"This guide is intended to help farmers estimate the breakeven price needed to justify harvesting a corn crop as silage rather than for grain. The estimated breakeven price of the standing crop becomes the corn farmer's lower boundary price for negotiations with a livestock producer wishing to purchase and harvest a field as silage. The value of silage delivered to storage accounts for harvest and transportation costs incurred by either the crop or livestock farmer. The livestock farmer's interest is in knowing the cost of silage (on a dry matter basis) delivered to the feed bunk, after accounting for storage losses and shrink. Livestock farmers compare silage's delivered cost per ton against costs of other feedstuffs."--First page.Revised by: Ray Massey (Professor, Agricultural Business and Policy Extension), Joe Horner (State Specialist, Agricultural Business and Policy Extension)New 10/2003; Revised 12/2020Includes bibliographical reference

Industrial hemp for grain and fiber planning budget

"This guide presents information useful to farmers considering industrial hemp production for grain and fiber in the 2021 crop year. Table 1 presents income and cost estimates for industrial hemp grain and fiber production in Missouri based on prices in October 2020. Information from Missouri and other hemp growing states have been used to develop this budget and production assumptions used in this budget may not describe all production activities. Assumptions are summarized in Tables 2 and 3. Assumed production is 40 acres. A sensitivity analysis showing the impact on profit of various yield and price combinations for grain is included in Table 4. Table 5 presents a sensitivity analysis of various yields for both fiber and grain assuming known prices for each."--First page.Written by: Ray Massey (Professor, Agricultural Business and Policy Extension), Joe Horner (State Specialist, Agricultural Business and Policy Extension)New 10/19; Revised 10/2

Pricing corn silage

"This guide is intended to help farmers estimate the breakeven price needed to justify harvesting a corn crop as silage rather than for grain. The estimated breakeven price of the standing crop becomes the corn farmer's lower boundary price for negotiations with a livestock producer wishing to purchase and harvest a field as silage. The value of silage delivered to storage accounts for harvest and transportation costs incurred by either the crop or livestock farmer."--Page 1.Revised by Ray Massey (Professor, Agricultural Business and Policy Extension), Joe Horner (State Specialist, Agricultural Business and Policy Extension)Includes bibliographical reference

Mexico Beckons Mid-American Small Business: Prospect for University Centered Export Trade Assistance*

Midwestern small business firms are being aggressively beckoned to enter the rapidly growing Mexican economy. With the recent passage of the North American Free Trade Agreement (NAFTA), Mexico is ready to start spending to upgrade its quality of life. Small business firms with products that can help with these improvements are being urged to scour Mexico for trade leads. This study reports on a survey of 1,104 midwest-central small and medium size manufacturing firms who find it difficult to take advantage of the export opportunity that Mexico may offer them. Among the respondents, 287firms are in industries classified by the U.S. Department of Commerce as having the greatest potential for rapid growth in export to Mexico. Characteristics of survey firms are presented along with their managerial and technical assistance needs. The prospect for a university-based export assistance center is explored along with the operational dimensions for such an agency partnership

Industrial hemp for grain planning budget

"This guide presents information useful to farmers considering industrial hemp production for grain in the 2021 crop year. Table 1 presents income and cost estimates for industrial hemp grain production in Missouri based on prices in October 2020. Assumptions are summarized in Tables 2 and 3. Assumed production is 40 acres. Several assumptions used in this budget are from states such as Kentucky, Tennessee, and North Dakota that have recently grown hemp. Farmers should understand that these assumptions may not fit Missouri production due to lack of Missouri data and different growing conditions in areas where data has been collected. A sensitivity analysis showing the impact on profit of various yields and market prices is included in Table 4."--First page.Written by: Ray Massey (Professor, Agricultural Business and Policy Extension), Joe Horner (State Specialist, Agricultural Business and Policy Extension)New 10/19; Revised 10/2