Experience in Action: Incorporating Somatic Education into the General Music Classroom

This thesis seeks to introduce principles of somatic education to children grades kindergarten through grade five in order to prevent future music-related injury and to promote free, wholebody movement. To do so, the principals of somatic education, specifically those of the Alexander Technique and Body Mapping, were combined with the current general music pedagogies by altering pre-existing lesson plans. Alterations include a variety of methods of integration, including, but not limited to: teaching about and exploring the different parts of the body, understanding the relationships between seemingly unrelated body parts, and engaging in student-led constructive rest in the semi-supine position. Somatic education pairs fluidly with the general music curriculum, as the primary pedagogues already incorporate movement and body awareness into their lessons. By pairing these lessons with an education about the body, students can move in a manner that relies on the skeletal structure rather than muscular effort, thus preventing common pains and music-related performance injuries

Circadian Entrainment Triggers Maturation of Human In Vitro Islets

Stem-cell-derived tissues could transform disease research and therapy, yet most methods generate functionally immature products. We investigate how human pluripotent stem cells (hPSCs) differentiate into pancreatic islets in vitro by profiling DNA methylation, chromatin accessibility, and histone modification changes. We find that enhancer potential is reset upon lineage commitment and show how pervasive epigenetic priming steers endocrine cell fates. Modeling islet differentiation and maturation regulatory circuits reveals genes critical for generating endocrine cells and identifies circadian control as limiting for in vitro islet function. Entrainment to circadian feeding/fasting cycles triggers islet metabolic maturation by inducing cyclic synthesis of energy metabolism and insulin secretion effectors, including antiphasic insulin and glucagon pulses. Following entrainment, hPSC-derived islets gain persistent chromatin changes and rhythmic insulin responses with a raised glucose threshold, a hallmark of functional maturity, and function within days of transplantation. Thus, hPSC-derived tissues are amenable to functional improvement by circadian modulation

Comparative Study of Active Flow Control Strategies for Lift Enhancement of a Simplified High-Lift Configuration

Numerical simulations have been performed for a simplified high-lift (SHL) version of the Common Research Model (CRM) configuration, where the Fowler flaps of the conventional high-lift (CRM-HL) configuration are replaced by a set of simple hinged flaps. These hinged flaps are equipped with integrated modular active flow control (AFC) cartridges on the suction surface, and the resulting geometry is known as the CRM-SHL-AFC configuration. The main objective is to make use of AFC devices on the CRM-SHL-AFC configuration to recover the aerodynamic performance (lift) of the CRM-HL configuration. In the current paper, a Lattice Boltzmann method-based computational fluid dynamics (CFD) code, known as PowerFLOWQ is used to simulate the entire flow field associated with the CRM-SHL-AFC configuration equipped with several different types of AFC devices. The transonic version of the PowerFLOWQ code that has been validated for high speed flows is used to accurately simulate the flow field generated by the high-momentum actuators required to mitigate reversed flow regions on the suction surfaces of the main wing and the flap. The numerical solutions predict the expected trends in aerodynamic forces as the actuation levels are increased. More efficient AFC systems and actuator arrangements emerged based on the parametric studies performed prior to a Fall 2018 wind tunnel test. Preliminary comparisons of the numerical solutions for lift and surface pressures are presented here with the experimental data, demonstrating the usefulness of CFD for predicting the flow field and lift characteristics of AFC-enabled high-lift configurations

Wind Tunnel Testing of Active Flow Control on High-Lift Common Research Model

A 10%-scale high-lift version of the Common Research Model (CRM-HL) and an Active Flow Control (AFC) version of the model equipped with a simple-hinged flap (CRM-SHLAFC) were successfully tested. The tests were performed in the 14- by 22-Foot Subsonic Tunnel (14x22) at the NASA Langley Research Center (LaRC). The CRM-HL has a set of 37 inboard and outboard single-element Fowler flaps. The CRM-SHL-AFC has a set of 50 inboard and 55 outboard simple-hinged flaps equipped with integrated modular AFC cartridges on the flap shoulder. Both high-lift configurations share the same 30 slats and engine nacelle. Three new types of AFC devices were examined: the Double-Row Sweeping Jets (DRSWJ), the Alternating Pulsed Jets (APJ), and the High Efficiency Low Power (HELP) actuators. The DRSWJ and the APJ actuators used two rows of unsteady jets, whereas the HELP actuators used a combination of unsteady and steady jets, to overcome strong adverse pressure gradients while minimizing the mass flow usage. Nozzle pressure ratio, mass flow consumption and the power coefficient, which takes account of both supply air pressure and mass flow usage for the actuators, were used for judging the performance efficiency of the AFC devices. A prestall lift performance degradation for the CRM-HL configuration was resolved with a properly placed nacelle chine. The configuration with nacelle chine was chosen as the representative reference conventional high-lift case for comparison with the CRMSHL- AFC. The AFC-induced lift coefficient increment (DCL) was maintained for the entire lift curve over the CRM-SHL-AFC case with no AFC for almost all flow-control cases examined. The lift curve of the reference CRM-HL have a slightly steeper slope compared to those of the CRM-SHL-AFC configurations. The HELP actuation concept was extremely effective in controlling flow separation in the linear region of the curves comparing lift coefficient to mass flow rate. The HELP actuation achieved a targeted DCL of 0.50 using a moderate amount of mass flow and supply air pressure. The CRM-SHL-AFC configuration equipped with HELP actuation was able to match or exceed the lift performance of the reference conventional high-lift configuration (i.e., CRM-HL equipped with a nacelle chine), thus meeting the NASA Advanced Air Transport Technology (AATT) project goal

Overexpression of Human Fatty Acid Transport Protein 2/Very Long Chain Acyl-CoA Synthetase 1 (FATP2/Acsvl1) Reveals Distinct Patterns of Trafficking of Exogenous Fatty Acids

In mammals, the fatty acid transport proteins (FATP1 through FATP6) are members of a highly conserved family of proteins, which function in fatty acid transport proceeding through vectorial acylation and in the activation of very long chain fatty acids, branched chain fatty acids and secondary bile acids. FATP1, 2 and 4, for example directly function in fatty acid transport and very long chain fatty acids activation while FATP5 does not function in fatty acid transport but activates secondary bile acids. In the present work, we have used stable isotopically labeled fatty acids differing in carbon length and saturation in cells expressing FATP2 to gain further insights into how this protein functions in fatty acid transport and intracellular fatty acid trafficking. Our previous studies showed the expression of FATP2 modestly increased C16:0-CoA and C20:4-CoA and significantly increased C18:3-CoA and C22:6-CoA after 4hr. The increases in C16:0-CoA and C18:3-CoA suggest FATP2 must necessarily partner with a long chain acyl CoA synthetase (Acsl) to generate C16:0-CoA and C18:3-CoA through vectorial acylation. The very long chain acyl CoA synthetase activity of FATP2 is consistent in the generation of C20:4-CoA and C22:6-CoA coincident with transport from their respective exogenous fatty acids. The trafficking of exogenous fatty acids into phosphatidic acid (PA) and into the major classes of phospholipids (phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositol (PI), and phosphatidyserine (PS)) resulted in distinctive profiles, which changed with the expression of FATP2. The trafficking of exogenous C16:0 and C22:6 into PA was significant where there was 6.9- and 5.3-fold increased incorporation, respectively, over the control; C18:3 and C20:4 also trended to increase in the PA pool while there were no changes for C18:1 and C18:2. The trafficking of C18:3 into PC and PI trended higher and approached significance. In the case of C20:4, expression of FATP2 resulted in increases in all four classes of phospholipid, indicating little selectivity. In the case of C22:6, there were significant increases of this exogenous fatty acids being trafficking into PC and PI. Collectively, these data support the conclusion that FATP2 has a dual function in the pathways linking the transport and activation of exogenous fatty acids. We discuss the differential roles of FATP2 and its role in both fatty acid transport and fatty acid activation in the context of lipid homeostasis

Mechanistic Studies of the Long Chain Acyl-CoA Synthetase Faa1p from \u3ci\u3eSaccharomyces cerevisiae\u3c/i\u3e

Long chain acyl-CoA synthetase (ACSL; fatty acid CoA ligase: AMP forming; EC 6.2.1.3) catalyzes the formation of acyl-CoA through a process, which requires fatty acid, ATP and coenzymeA as substrates. In the yeast Saccharomyces cerevisiae the principal ACSL is Faa1p (encoded by the FAA1 gene). The preferred substrates for this enzyme are cis-monounsaturated long chain fatty acids. Our previous work has shown Faa1p is a principal component of a fatty acid transport/activation complex that also includes the fatty acid transport protein Fat1p. In the present work hexameric histidine tagged Faa1p was purified to homogeneity through a two-step process in the presence of 0.1% η-dodecyl-β-maltoside following expression at 15°C in Escherichia coli. In order to further define the role of this enzyme in fatty acid transport-coupled activation (vectorial acylation), initial velocity kinetic studies were completed to define the kinetic parameters of Faa1p in response to the different substrates and to define mechanism. These studies showed Faa1p had a Vmax of 158.2 nmol/ min/mg protein and a Km of 71.1μM oleate. When the concentration of oleate was held constant at 50μM, the Km for CoA and ATP were 18.3μM and 51.6μM respectively. These initial velocity studies demonstrated the enzyme mechanism for Faa1p was Bi Uni Uni Bi Ping Pong

Pseudomonas aeruginosa Magnesium Transporter MgtE Inhibits Type III Secretion System Gene Expression by Stimulating rsmYZ Transcription

Pseudomonas aeruginosa causes numerous acute and chronic opportunistic infections in humans. One of its most formidable weapons is a type III secretion system (T3SS), which injects powerful toxins directly into host cells. The toxins lead to cell dysfunction and, ultimately, cell death. Identification of regulatory pathways that control T3SS gene expression may lead to the discovery of novel therapeutics to treat P. aeruginosa infections. In a previous study, we found that expression of the magnesium transporter gene mgtE inhibits T3SS gene transcription. MgtE-dependent inhibition appeared to interfere with the synthesis or function of the master T3SS transcriptional activator ExsA, although the exact mechanism was unclear. We now demonstrate that mgtE expression acts through the GacAS two-component system to activate rsmY and rsmZ transcription. This event ultimately leads to inhibition of exsA translation. This inhibitory effect is specific to exsA as translation of other genes in the exsCEBA operon is not inhibited by mgtE Moreover, our data reveal that MgtE acts solely through this pathway to regulate T3SS gene transcription. Our study reveals an important mechanism that may allow P. aeruginosa to fine-tune T3SS activity in response to certain environmental stimuli.IMPORTANCE The type III secretion system (T3SS) is a critical virulence factor utilized by numerous Gram-negative bacteria, including Pseudomonas aeruginosa, to intoxicate and kill host cells. Elucidating T3SS regulatory mechanisms may uncover targets for novel anti-P. aeruginosa therapeutics and provide deeper understanding of bacterial pathogenesis. We previously found that the magnesium transporter MgtE inhibits T3SS gene transcription in P. aeruginosa In this study, we describe the mechanism of MgtE-dependent inhibition of the T3SS. Our report also illustrates how MgtE might respond to environmental cues, such as magnesium levels, to fine-tune T3SS gene expression

Human Fatty Acid Transport Protein 2a/Very Long Chain Acyl-CoA Synthetase 1 (FATP2a/Acsvl1) Has a Preference in Mediating the Channeling of Exogenous n-3 Fatty Acids into Phosphatidylinositol

The trafficking of fatty acids across the membrane and into downstream metabolic pathways requires their activation to CoA thioesters. Members of the fatty acid transport protein/ very long chain acyl-CoA synthetase (FATP/Acsvl) family are emerging as key players in the trafficking of exogenous fatty acids into the cell and in intracellular fatty acid homeostasis.We have expressed two naturally occurring splice variants of human FATP2 (Acsvl1) in yeast and 293T-REx cells and addressed their roles in fatty acid transport, activation, and intracellular trafficking. Although both forms (FATP2a (Mr 70,000) and FATP2b (Mr 65,000 and lacking exon3, which encodes part of the ATP binding site)) were functional in fatty acid import, only FATP2a had acyl-CoA synthetase activity, with an apparent preference toward very long chain fatty acids. To further address the roles of FATP2a or FATP2b in fatty acid uptake and activation, LCMS/ MS was used to separate and quantify different acyl-CoA species (C14–C24) and to monitor the trafficking of different classes of exogenous fatty acids into intracellular acyl-CoA pools in 293T-REx cells expressing either isoform. The use of stable isotopically labeled fatty acids demonstrated FATP2a is involved in the uptake and activation of exogenous fatty acids, with a preference toward n-3 fatty acids (C18:3 and C22:6). Using the same cells expressing FATP2a or FATP2b, electrospray ionization/MS was used to follow the trafficking of stable isotopically labeled n-3 fatty acids into phosphatidylcholine and phosphatidylinositol. The expression of FATP2a resulted in the trafficking of C18:3-CoA and C22:6-CoA into both phosphatidylcholine and phosphatidylinositol but with a distinct preference for phosphatidylinositol. Collectively these data demonstrate FATP2a functions in fatty acid transport and activation and provides specificity toward n-3 fatty acids in which the corresponding n-3 acyl-CoAs are preferentially trafficked into acyl-CoA pools destined for phosphatidylinositol incorporation

Numerical Simulation of a Simplified High-Lift CRM Configuration Embedded with Fluidic Actuators

Numerical simulations have been performed for a simplified high-lift configuration that is representative of a modern transport airplane. This configuration includes a leading-edge slat, fuselage, wing, nacelle-pylon and a simple hinged flap. The suction surface of the flap is embedded with multiple rows of fluidic actuators to reduce the extent of reversed flow regions and improve the aerodynamic performance of the configuration with flap in a deployed state. In the current paper, a Lattice Boltzmann Method based high-fidelity computational fluid dynamics (CFD) code, known as PowerFLOW is used to simulate the entire flow field associated with this configuration, including the flow inside the actuators. A fully compressible version of the PowerFLOW code that has been validated for high speed flows is used for the present simulations to accurately represent the transonic flow regimes that are encountered in the flow field generated by the actuators operating at higher mass flow (momentum) rates required to mitigate reverse flow regions on the suction surfaces of the main wing and the flap. The numerical solutions predict the expected trends in aerodynamic forces as the actuation levels are increased. More efficient active flow control (AFC) systems and actuator arrangement for lift augmentation are emerging based on the parametric studies conducted here prior to wind tunnel tests. These numerical solutions will be compared with experimental data, once such data becomes available

Biogenesis of the mitochondrial phosphate carrier

The mitochondrial phosphate carrier (PiC) is a member of the family of inner-membrane carrier proteins which are generally synthesized without a cleavable presequence. Surprisingly, the cDNA sequences of bovine and rat PiC suggested the existence of an amino-terminal extension sequence in the precursor of PiC. By expressing PiC in vitro, we found that PiC is indeed synthesized as a larger precursor. This precursor was imported and proteolytically processed by mitochondria, whereby the correct amino-terminus of the mature protein was generated. Import of PiC showed the characteristics of mitochondrial protein uptake, such as dependence on ATP and a membrane potential and involvement of contact sites between mitochondrial outer and inner membranes. The precursor imported in vitro was correctly assembled into the functional form, demonstrating that the authentic import and assembly pathway of PiC was reconstituted when starting with the presequence-carrying precursor. These results are discussed in connection with the recently postulated role of PiC as an import receptor located in the outer membrane