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At least 1,135 records · Page 63

Activation of galanin receptor 1 inhibits locus coeruleus neurons via GIRK channels

The noradrenergic neurons of the locus coeruleus (LC) are associated with various brain functions and psychiatric disorders, such as addiction and depression. It has been shown that neuropeptide galanin (GAL) inhibits neuronal excitability in LC, but the mechanisms remain unclear. In the present study, we investigated the ionic and signal transduction mechanisms underlying inhibitory effect of GAL on LC neurons using whole-cell patch clamp recording in rat brain slices. Bath application of GAL decreased the spontaneous firings and induced a dose-dependent hyperpolarization of LC neurons and this effect was attenuated by knockdown of Galr1, but not Galr2, confirming that mainly GALR1 mediates the inhibition effect of GAL. The inhibitory effect of GAL was also blocked by treatments of pertussis toxin (PTX), GTP-γ-s or GDP-β-s, respectively, indicating that the functions of PTX sensitive G{sub i/o} protein are required for GAL-induced hyperpolarization. Moreover, the blockers of GIRK (tertiapin-Q or SCH2 3390 hydrochloride) attenuated the GAL response while blocker of BK/SK/K{sub ATP} channels or TASK-1/3 channels did not affect it significantly, suggesting that GIRK channels play an important role in GAL-induced hyperpolarization in LC neurons. Taken together, the inhibitory effect of GAL on LC neurons is mediated by GALR1 via PTX-sensitive G{sub i/o} proteins, which activate GIRK channels.

60 APPLIED LIFE SCIENCES↗

Fibromodulin modulates myoblast differentiation by controlling calcium channel

Fibromodulin (FMOD) is a proteoglycan present in extracellular matrix (ECM). Based on our previous findings that FMOD controls myoblast differentiation by regulating the gene expressions of collagen type I alpha 1 (COL1α1) and integral membrane protein 2 A (Itm2a), we undertook this study to investigate relationships between FMOD and calcium channels and to understand further the mechanism by which they control myoblast differentiation. Gene expression studies and luciferase reporter assays showed FMOD affected calcium channel gene expressions by regulating calcium channel gene promoter, and patch-clamp experiments showed both L- and T-type calcium channel currents were almost undetectable in FMOD knocked down cells. In addition, gene knock-down studies demonstrated the COL1α1 and Itm2a genes both regulate the expressions of calcium channel genes. Studies using a cardiotoxin-induced mouse muscle injury model demonstrated calcium channels play important roles in the regeneration of muscle tissue, possibly by promoting the differentiation of muscle stem cells (MSCs). Summarizing, the study demonstrates ECM components secreted by myoblasts during differentiation provide an essential environment for muscle differentiation and regeneration.

60 APPLIED LIFE SCIENCES↗

Natronomonas pharaonis halorhodopsin Ser81 plays a role in maintaining chloride ions near the Schiff base

Highlights: • S81A mutant of NpHR lose light-induced photocurrent. • The direction of the Thr126 side chain was fixed by a hydroxyl group of Ser81. • S81A mutation decrease the interaction between Thr126 and a chloride ion. • Ser81 plays a key role in chloride migration. Optogenetic technologies have often been used as tools for neuronal activation or silencing by light. Natronomonas pharaonis halorhodopsin (NpHR) is a light-driven chloride ion pump. Upon light absorption, a chloride ion passes through the cell membrane, which is accompanied by the temporary binding of a chloride ion with Thr126 at binding site-1 (BS1) near the protonated Schiff base in NpHR. However, the mechanism of stabilization of the binding state between a chloride ion and BS1 has not been investigated. Therefore, to identify a key component of the chloride ion transport pathway as well as to acquire dynamic information about the chloride ion-BS1 binding state, we performed a rough analysis of the chloride ion pathway shape followed by molecular dynamics (MD) simulations for both wild-type and mutant NpHR structures. The MD simulations showed that the hydrogen bond between Thr126 and the chloride ion was retained in the wild-type protein, while the chloride ion could not be retained at and tended to leave BS1 in the S81A mutant. We found that the direction of the Thr126 side chain was fixed by a hydroxyl group of Ser81 through a hydrogen bond and that Thr126 bound to a chloride ion in the wild-type protein, while this interaction was lost in the S81A mutant, resulting in rotation of the Thr126 side chain and reduction in the interaction between Thr126 and a chloride ion. To confirm the role of S81, patch clamp recordings were performed using cells expressing NpHR S81A mutant protein. Considered together with the results that the NpHR S81A-expressing cells did not undergo hyperpolarization under light stimulation, our results indicate that Ser81 plays a key role in chloride migration. Our findings might be relevant to ongoing clinical trials using optogenetic gene therapy in blind patients.

60 APPLIED LIFE SCIENCES↗

Dual regulation of hEAG1 channels by phosphatidylinositol 4,5-bisphosphate

Highlights: • hEAG1 channels are PIP{sub 2} sensitive. • hEAG1 channels are inhibited either by PIP{sub 2} depletion or loading. • Optimum membrane PIP{sub 2} level is required for the proper functioning of hEAG1 channels. The ether-à-go-go1 (EAG1, Kv10.1) K{sup +} channel is a member of the voltage-gated K{sup +} channel family mainly expressed in the central nervous system and cancer cells. Membrane lipids regulate several voltage-gated K{sup +} channels but their influence on EAG1 channels has been poorly explored. Here we have studied the regulation of hEAG1 channels by phosphatidylinositol 4,5-bisfofate (PIP{sub 2}) by using different strategies to manipulate the levels of this lipid, and the patch clamp technique. We found that depletion of endogenous PIP{sub 2} by activation of the voltage-sensing phosphatase from Danio rerio (Dr-VSP) or the human muscarinic type-1 receptor (hM1R) inhibits hEAG1 currents; however, the application of exogenous PIP{sub 2} to increase the level of this lipid on the plasma membrane, also induced an inhibition of hEAG1. In summary, our results indicate that PIP{sub 2} have dual effects on hEAG1 channels and its action as activator or inhibitor depends on its initial level on the plasma membrane.

60 APPLIED LIFE SCIENCES↗

Acetylation of lysine residues in the recombinant nucleoprotein and VP40 matrix protein of Zaire Ebolavirus by eukaryotic histone acetyltransferases

Acetylation of histones and other proteins plays crucial roles in transcriptional regulation, chromatin organization, and other biological processes. It has been recently reported that the nucleoprotein (NP) of influenza virus is acetylated in infected cells, and this modification contributes to the RNA polymerization activity of the virus. As the influenza virus, the Ebolavirus contains single-stranded negative-sense RNA as its viral genome, which interacts with NP and other viral proteins. In this study, we performed a series of biochemical experiments and revealed that the recombinant Ebolavirus NP and the viral matrix protein VP40, which binds with NP, were acetylated by eukaryotic histone acetyltransferases, such as P300/CREB-binding protein (P300/CBP) and P300/CBP-associated factor (PCAF), in vitro. Mass spectrometry was used to identify the lysine residues that were potential acetylation targets in NP and VP40. The identified lysine residues in NP were located in the RNA-binding cleft and the VP35-binding domain. Potentially acetylated lysine targets in VP40 were identified in the basic patch, which is necessary for constructing oligomers. These results suggest that the acetylation of these lysine residues is involved in the interactions between viral proteins.

60 APPLIED LIFE SCIENCES↗

An efficient method to integrate polynomials over polytopes and curved solids

Here in this paper, we present an efficient approach to compute the integral of monomials and polynomials over polyhedra and regions defined by parametric curved boundary surfaces. We use Euler's theorem for homogeneous functions in combination with Stokes's theorem to reduce the integration of a monomial over a three-dimensional solid to its boundary. If the solid is a polytope, through a recursive application of these theorems, the integral is further reduced to just the evaluation of the monomial and its derivatives at the vertices of the polytope. The present approach is simpler than existing techniques that rely on repeated use of the divergence theorem, which require the antiderivative of the monomials and the projection of these functions onto hyperplanes. For convex and nonconvex polytopes, our approach does not introduce any approximation for the integration of monomials. For curved solid regions bounded by surfaces that admit a parameterization, the same approach yields simplified formulas to compute the integral of any homogeneous function, including monomials. For surfaces parameterized by polynomial surfaces (such as Bezier surface triangles and B-spline patches), the method yields machine-precision accuracy for the volumetric integration of monomials with an appropriate quadrature rule. Numerical examples over regions bounded by polynomial surfaces and rational surfaces are presented to establish the accuracy and efficiency of the method.

97 MATHEMATICS AND COMPUTING↗

The Spectrin-Actin-Based Periodic Cytoskeleton as a Conserved Nanoscale Scaffold and Ruler of the Neural Stem Cell Lineage

Through three-dimensional STORM super-resolution microscopy, we resolve the spectrin-actin-based membrane cytoskeleton of neural stem cells (NSCs) and NSC-derived neurons, astrocytes, and oligodendrocytes. We show that undifferentiated NSCs are capable of forming patches of locally periodic, one-dimensional (1D) membrane cytoskeleton with ~180 nm periodicity. Such periodic structures become increasingly ordered and long-ranging as the NSCs mature into terminally differentiated neuronal and glial cell types, and, during this process, distinct 1D periodic “strips” dominate the flat 2D membranes. Moreover, we report remarkable alignment of the periodic cytoskeletons between abutting cells at axon-axon and axon-oligodendrocyte contacts and identify two adhesion molecules, neurofascin and L1CAM, as candidates to drive this nanoscale alignment. We thus show that a conserved 1D periodic membrane cytoskeletal motif serves as a nanoscale scaffold and ruler to mediate the physical interactions between cell types of the NSC lineage.

59 BASIC BIOLOGICAL SCIENCES↗

Dynamic flux surrogate-based partitioned methods for interface problems

Loosely coupled partitioned methods for multiphysics problems treat each subproblem as a separate entity and advance them independently in time. In so doing these methods enable code reuse, increase concurrency and provide a convenient framework for plug-and-play multiphysics simulations. However, mathematically loosely coupled schemes are equivalent to a single step of an iterative solution method, which can compromise their accuracy and stability. We present a new data-driven partitioned method for coupled parametric PDEs that can improve upon the accuracy of traditional loosely coupled methods without incurring a performance penalty. To that end, we replace conventional field transfers across the interface by a surrogate for the dynamics of the interface flux exchanged between the subdomains. To develop this surrogate we apply dynamic mode decomposition to a non-standard staggered-in-time state, comprising the interface flux and small solution patches near the interface. The new approach shifts the main computational burden to an offline training phase, whereas application of the surrogate in the online phase amounts to a single matrix–vector multiplication. In conclusion, we provide stability analysis of the surrogate-based partitioned scheme and include numerical results that demonstrate its potential.

Dynamic mode decomposition (DMD)↗

A splice method for local-to–nonlocal coupling of weak forms

Here, we propose a method to couple local and nonlocal diffusion models. By inheriting desirable properties such as patch tests, asymptotic compatibility and unintrusiveness from related splice and optimization-based coupling schemes, it enables the use of weak (or variational) formulations, is computationally efficient and straightforward to implement. We prove well-posedness of the coupling scheme and demonstrate its properties and effectiveness in a variety of numerical examples.

97 MATHEMATICS AND COMPUTING↗

Adaptive workflow for simulation of RF heaters

Accurate RF (Radio Frequency) simulations of fusion systems like ITER require the definition of high-fidelity analysis geometries that include detailed antenna, reactor wall, and physics regions. Here, this paper will describe a workflow for the execution of adaptive high-performance simulations of RF fusion systems. In this workflow, the simulation input consists of a CAD model attributed with the needed analysis attributes. The analysis mesh is automatically generated and the analysis steps are executed using the time-harmonic Maxwell's equations solved using high-order Nédélec finite elements. A patch recovery-based error estimator is used to drive a conforming mesh adaptation procedure.

97 MATHEMATICS AND COMPUTING↗

Assessment of the hydromechanical higher-order MPM for the simulation of geotechnical problems

The Material Point Method (MPM) has been increasingly used to simulate large strain deformations. Linear interpolation functions are commonly used to perform the spatial integration. It is well-known that the discontinuities in the interpolation function derivatives induce shock-like artifacts known as ‘cell-crossing’ error. These errors compound with volumetric locking errors when used with hydromechanical formulations for porous media, where different velocity fields are used for each phase. The capabilities of higher-order MPM frameworks have not been explored for real-scale geotechnical problems. As such, this paper aims to assess, validate, and further discuss a higher-order B-spline MPM (BS-MPM) framework. First, the BS-MPM framework is verified against the large-strain oedometer consolidation problem. Second, the framework is validated against a real-scale slope failure experiment triggered by pore water pressure recharge. Landslide features that are captured using the higher-order framework are specifically highlighted, and results (e.g., pore water pressure and deformation) are validated with field measurements. A generally convergent numerical solution is observed when using cubic interpolation functions. Third, a footing penetration problem is simulated using the multi-patch BS-MPM. Trends are examined with respect to penetration velocity and variation in hydraulic conductivity. The BS-MPM framework ultimately presents a stabilized numerical solution that captures plausible hydromechanical interaction trends important in geotechnical engineering applications.

36 MATERIALS SCIENCE↗

Effects of Asperity Distribution on Fluid Flow and Induced Seismicity During Deep Geothermal Exploitation

This work investigates the injection-induced seismic response of a heterogeneous fault plane, featuring low-permeability asperities embedded into a high-permeability damage zone. We simulate the pressure evolution with a hydrogeological simulator, accounting for the heterogeneous fault plane. Seismicity occurs then on the asperities, represented as unstable patches reactivating by means of the Mohr-Coulomb criterion. The hydrological and seismic modules are implicitly coupled to account for effects of asperity reactivation on the permeability. Results show that permeability changes may cause at a later time a change in seismicity propagation. Finally, we also investigated such effects by varying the density of asperities.

58 GEOSCIENCES↗

Assessing land take by urban development and its impact on carbon storage: Findings from two case studies in Italy

Highlights: • We tested a new methodology for monitoring land take and its effects on C storage. • The ecological impact of urban growth derives from the previous land use. • C loss increases with the naturalness of the territory. • Different urban assets may imply different forms of land take containment. Land take due to urbanization triggers a series of negative environmental impacts with direct effects on quality of life for people living in cities. Changes in ecosystem services are associated with land take, among which is the immediate C loss due to land use conversion. Land use change monitoring represents the first step in quantifying land take and its drivers and impacts. To this end, we propose an innovative methodology for monitoring land take and its effects on ecosystem services (in particular, C loss) under multi-scale contexts. The devised approach was tested in two areas with similar sizes, but different land take levels during the time-span 1990–2008 in Central Italy (the Province of Rome and the Molise Region). The estimates of total coverage of built up areas were calculated using point sampling. The area of the urban patches including each sampling point classified as built up areas in the year 1990 and/or in the year 2008 is used to estimate total abundance and average area of built up areas. Biophysical and economic values for carbon loss associated with land take were calculated using InVEST. Although land take was 7–8 times higher in the Province of Rome (from 15.1% in 1990 to 20.4% in 2008) than in Molise region, our findings show that its relative impact on C storage is higher in the latter, where the urban growth consistently affects not only croplands but also semi-natural land uses such as grasslands and other wooded lands. The total C loss due to land take has been estimated in 1.6 million Mg C, corresponding to almost 355 million €. Finally, the paper discusses the main characteristics of urban growth and their ecological impact leading to risks and challenges for future urban planning and land use policies.

54 ENVIRONMENTAL SCIENCES↗

Effects of triphenyltin on glycinergic transmission on rat spinal neurons

Highlights: • Effects of triphenyltin (TPT) were examined using ‘synaptic bouton’ preparation. • TPT did not change extrasynaptic glycine receptor-mediated responses. • TPT modulated glycinergic spontaneous and evoked synaptic responses. • TPT at high concentrations finally depleted glycine in the nerve terminal. • It is likely that TPT causes breaking down of neuronal excitability in wild creatures. Glycine is a fast inhibitory transmitter like γ-aminobutyric acid in the mammalian spinal cord and brainstem, and it is involved in motor reflex, nociception, and neuronal development. Triphenyltin (TPT) is an organometallic compound causing environmental hazard to many wild creatures. Our previous findings show that TPT ultimately induces a drain and/or exhaustion of glutamate in excitatory presynaptic nerve terminals, resulted in blockage of neurotransmission as well as methylmercury. Therefore, we have investigated the neurotoxic mechanism how TPT modulates inhibitory glycinergic transmission in the synaptic bouton preparation of rat isolated spinal neurons using a patch clamp technique. TPT at environmentally relevant concentrations (3–300 nM) significantly increased the number of frequency of glycinergic spontaneous and miniature inhibitory postsynaptic currents (sIPSC and mIPSC) without affecting the current amplitude and decay time. The TPT effects were also observed in external Ca{sup 2+}-free solution containing tetrodotoxin (TTX) but removed in Ca{sup 2+}-free solution with both TTX and BAPTA-AM (Ca{sup 2+} chelator). On the other hand, the amplitude of glycinergic evoked inhibitory postsynaptic currents (eIPSCs) increased with decreasing failure rate (Rf) and paired pulse ratio (PPR) in the presence of 300 nM TPT. At a high concentration (1 µM), TPT completely blocked eIPSCs after a transient facilitation. Overall, these results suggest that TPT directly acts transmitter-releasing machinery in glycinergic nerve terminals. Effects of TPT on the nerve terminals releasing fast transmitters were greater in the order of glycinergic > glutamatergic > GABAergic ones. Thus, TPT is supposed to cause a strong synaptic modulations on glycinergic neurotransmission in wild creatures.

54 ENVIRONMENTAL SCIENCES↗

The geophysics, geology and mechanics of slow fault slip

We report that modern geodetic and seismologic observations describe the behavior of fault slip over a vast range of spatial and temporal scales. Slip at sub-seismogenic speeds is evident from top to bottom of lithospheric faults and plays an important role throughout the earthquake cycle. Where earthquakes and tremor accompany slow slip, they help illuminate the spatiotemporal evolution of fault slip. Geophysical subsurface imaging and geologic field studies provide information about suitable environments of slow slip. In particular, exhumed fault and shear zones from various depths reveal the importance of multiple deformation processes and fault-zone structures. Most geologic examples feature frictionally weak and velocity-strengthening materials, well-developed mineral fabrics, and abundant veining indicative of near-lithostatic fluid pressure. To produce transient slow slip events and tremor, in addition to the presence of high-pressure fluids a heterogeneous fault-zone structure, composition, and/or metamorphic assemblage may be needed. Laboratory and computational models suggest that velocity-weakening slip patches smaller than a critical dimension needed for earthquake nucleation will also fail in slow slip events. Changes in fluid pressure or slip rate can cause a fault to transition between stable and unstable fault slip behavior. Lastly, future interdisciplinary investigations of slow fault slip, directly integrating geophysical, geological and modeling investigations, will further improve our understanding of the dynamics of slow slip and aid in providing more accurate earthquake hazard characterizations.

58 GEOSCIENCES↗

Performance of multifunctional piezoelectric energy harvesting microgyroscopes with material degradation

Most microelectromechanical systems (MEMS) are made from brittle materials that are prone to cracking. Damage from the formation of cracks can occur anytime during the manufacturing process or due to mechanical loading of MEMS devices. Cracks can increase the structures flexibility and lead to changes in performance or total failure. Understanding the potential influences cracks in MEMS is necessary for the longevity of these systems. The energy harvesting microgyroscope studied in this work is susceptible to crack propagation in its silicon substrate and lead zirconate titanate piezoelectric patch. To this end, an electromechanical model is developed to capture the impacts of cracks on the performance of a damaged energy harvesting microgyroscope. Numerical methods are used to approximate the damage associated with distributed crack networks. Then, the static and dynamic responses of the damaged microgyroscope are analyzed for two possible multifunctional setups. Here, the current study revealed that cracks can significantly affect the static and dynamic behaviors of the microgyroscope energy harvester. Combinations of cracking and piezoelectric property degradation cause a decrease in the static pull-in voltage and shift the resonance frequencies of the system. Additionally, it is shown that the systems sensitivity to changes in the base rotation and energy harvesting capabilities vary widely depending on the severity of material degradation.

36 MATERIALS SCIENCE↗

Drivers and ecological impacts of a wildfire outbreak in the southern Appalachian Mountains after decades of fire exclusion

During one of the warmest and driest droughts of the last century, the southern Appalachian Mountains experienced a regional outbreak of over a dozen large wildfires in late fall of 2016. We provide a synthesis of long-term forest changes leading up to the 2016 wildfires, examine the climatic setting and patterns of burn severity in relationship to topography, and discuss the ecological and management implications of these and future fires. During the pre- and post-European settlement periods, frequent low- and mixed-severity wildfires interacted with complex topographic gradients and maintained heterogenous landscapes dominated by several species of oak (Quercus spp.), pine (Pinus spp.), and the American chestnut (Castanea dentata). Land-use changes associated with European settlement, loss of the American chestnut, and 20th century fire exclusion resulted in large scale shifts towards mesic, fire-intolerant species (i.e. mesophication). Wildfire activity began increasing in the early 1980s, but most fires in the region are small (<1,000 ha). In Fall of 2016, deciduous tree leaf fall occurred concurrently with a period of anomalously dry and warm weather, creating ideal conditions for fire ignition and spread. Thousands of ignitions across the region strained suppression resources and eight fires grew to greater than 5,000 ha. The 2016 fires were larger and burned more area than in the previous three decades combined. In one unique landscape setting, the Chimney Tops 2 Fire, a synoptic wind event drove extreme fire behavior and burned large, high-severity patches resulting in devastating effects in the wildland urban interface. However, immediate post-fire burn severity mosaics for other fires were composed primarily of low- (73 %) and moderate-severity (21 %) fire effects. High-severity fire comprised only 6 % of the area burned, and occurred mostly on steep upper slopes and ridges on south-facing aspects, reflecting the importance of bottom-up topographic drivers in this region. Although the fires will likely enhance biodiversity by restoring fire-dependent species and creating early seral habitat, invasions of non-native plant species, delayed mortality of mature pines and oaks, and rapid re-sprouting of pyrophyllic shrubs pose significant management challenges. Although similar large fire outbreaks may become more common under future climatic conditions, they are unlikely to reverse the effects of mesophication and significantly alter forest dynamics at broad spatial scales. Furthermore, the 2016 fires exposed the vulnerability of the region to wildfire during acute fall drought and demonstrate the potential ecological effects of future wildfires in mixed pine-hardwood landscapes of the southern Appalachians.

54 ENVIRONMENTAL SCIENCES↗