Corticosterone Treatment Results in Enhanced Release of Peptidergic Vesicles in Astrocytes via Cytoskeletal Rearrangements

While the effect of stress on neuronal physiology is widely studied, its effect on the functionality of astrocytes is not well understood. We studied the effect of high doses of stress hormone corticosterone, on two physiological properties of astrocytes, i.e., gliotransmission and interastrocytic calcium waves. To study the release of peptidergic vesicles from astrocytes, hippocampal astrocyte cultures were transfected with a plasmid to express pro-atrial natriuretic peptide (ANP) fused with the emerald green fluorescent protein (ANP.emd). The rate of decrease in fluorescence of ANP.emd on application of ionomycin, a calcium ionophore was monitored. Significant increase in the rate of calcium-dependent exocytosis of ANP.emd was observed with the 100 nM and 1 M corticosterone treatments for 3 h, which depended on the activation of the glucocorticoid receptor. ANP.emd tagged vesicles exhibited increased mobility in astrocyte culture upon corticosterone treatment. Increasing corticosterone concentrations also resulted in concomitant increase in the calcium wave propagation velocity, initiated by focal ATP application. Corticosterone treatment also resulted in increased GFAP expression and F-actin rearrangements. FITC-Phalloidin immunostaining revealed increased formation of cross linked F-actin networks with the 100 nM and 1 M corticosterone treatment. Alternatively, blockade of actin polymerization and disruption of microtubules prevented the corticosterone-mediated increase in ANP.emd release kinetics. This study reports for the first time the effect of corticosterone on gliotransmission via modulation of cytoskeletal elements. As ANP acts on both neurons and blood vessels, modulation of its release could have functional implications in neurovascular coupling under pathophysiological conditions of stress.

Switching and release dynamics of an electrostatically actuated MEMS switch under the influence of squeeze-film damping

This paper reports analytical modeling, simulation and experimental validation for switching and release times of an electrostatically actuated micromachined switch. Presented work is an extension of our earlier work [1] that analytically argued, and numerically and experimentally demonstrated, why pull-in time is larger that pull-up time when the actuation voltage is less than twice of the pull-in voltage. In this paper, switching dynamics is investigated under the influence of squeeze-film damping. Tests were performed on SOI (silicon-on-insulator) based parallel beams structures.Typical voltage requirement for actuation is in the range of 10-30 V. All the experiments were performed in normal atmospheric pressure. Measurement results confirm that the quality factor Q has appreciable effect on the release time compared to the switching time. The quality factor Q is extracted from the response measurement and compared with the ANSYS simulation result. In addition, the dynamic pull-in effect has also been studied and reported in this paper. A contribution of this work includes the effect of various phenomena such as squeeze-film damping, dynamic pull-in, and frequency pull-in effects on the switching dynamics of a MEMS switch.

Infection of Human Endothelial Cells by Japanese Encephalitis Virus: Increased Expression and Release of Soluble HLA-E

Japanese encephalitis virus (JEV) is a single stranded RNA virus that infects the central nervous system leading to acute encephalitis in children. Alterations in brain endothelial cells have been shown to precede the entry of this flavivirus into the brain, but infection of endothelial cells by JEV and their consequences are still unclear. Productive JEV infection was established in human endothelial cells leading to IFN-beta and TNF-alpha production. The MHC genes for HLA-A, -B, -C and HLA-E antigens were upregulated in human brain microvascular endothelial cells, the endothelial-like cell line, ECV 304 and human foreskin fibroblasts upon JEV infection. We also report the release/shedding of soluble HLA-E (sHLA-E) from JEV infected human endothelial cells for the first time. This shedding of sHLA-E was blocked by an inhibitor of matrix metalloproteinases (MMP). In addition, MMP-9, a known mediator of HLA solubilisation was upregulated by JEV. In contrast, human fibroblasts showed only upregulation of cell-surface HLA-E. Addition of UV inactivated JEV-infected cell culture supernatants stimulated shedding of sHLA-E from uninfected ECV cells indicating a role for soluble factors/cytokines in the shedding process. Antibody mediated neutralization of TNF-alpha as well as IFNAR receptor together not only resulted in inhibition of sHLA-E shedding from uninfected cells, it also inhibited HLA-E and MMP-9 gene expression in JEV-infected cells. Shedding of sHLA-E was also observed with purified TNF-alpha and IFN-beta as well as the dsRNA analog, poly (I:C). Both IFN-beta and TNF-alpha further potentiated the shedding when added together. The role of soluble MHC antigens in JEV infection is hitherto unknown and therefore needs further investigation.

Impact angle constraint guidance law using cubic splines for intercepting stationary targets

In this paper the cubic spline guidance law is presented for intercepting a stationary target at a desired impact angle. The guidance law is obtained from cubic spline curve based trajectory using an inverse method. The cubic spline t rajectory curve expresses the altitude as a cubic polynomial of the downrange. The guidance law is modified to achieve interception in the cases where impact angle is greater that or equal to 90◦. The guidance law is implemented in a feedback mode to maintain the desired impact angle and to reduce miss distance in the presence of lateral acceleration saturation and atmospheric distur- bances. The simulation results show that the guidance law fulfills all the requirements.

Protamine-Capped Mesoporous Silica Nanoparticles for Biologically Triggered Drug Release

The fabrication of a mesoporous silica nanoparticle (MSN)-protamine hybrid system (MSN-PRM) is reported that selectively releases drugs in the presence of specific enzyme triggers present in the proximity of cancer cells. The enzyme trigger involved is a protease called trypsin, which is overexpressed in certain specific pathological conditions, such as inflammation and cancer. Overexpression of trypsin is known to be associated with invasion, metastasis, and growth in several cancers, such as leukemia, colon cancer, and colorectal cancer. The current system (MSN-PRM) consists of an MSN support in which mesopores are capped with an FDA-approved peptide drug protamine, which effectively blocks the outward diffusion of the drug molecules from the mesopores of the MSNs. On exposure to the enzyme trigger, the protamine cap disintegrates, opening up the molecular gates and releasing the entrapped drug molecules. The system exhibits minimal premature release in the absence of the trigger and selectively releases the encapsulated drugs in the presence of the proteases secreted by colorectal cancer cells. The ability of the MSN-PRM particles to deliver anticancer drugs to colorectal cancer cells has also been demonstrated. The hydrophobic drug is released into cancer cells subsequent to disintegration of the protamine cap, resulting in cell death. Drug-induced cell death in colorectal cancer cells is significantly enhanced when the hydrophobic drug that is known to degrade in aqueous environments is encapsulated in the MSN-PRM system in comparison to the free drug (P < 0.05). The system, which shows good biocompatibility and selective drug release, is a promising platform for cancer specific drug delivery.

Polyanhydrides of Castor Oil-Sebacic Acid for Controlled Release Applications

A family of high molecular weight castor oil (CO)-based biodegradable polyanhydrides was synthesized by a catalyst-free melt-condensation reaction between prepolymers of CO and sebacic acid (SA). The structure of the polymers was characterized by H-1 NMR and Fourier transform infrared spectroscopy, which indicated the formation of the anhydride bond along the polymer backbone. Thermal analysis and X-ray diffraction confirmed the semicrystalline nature of the polymers. Incorporation of SA enhanced the crystallinity of the polymer. The hydrophobic nature of these polymers was revealed by contact angle goniometry. Water wettability decreased with increase in SA content. Compressive tests demonstrated a sharp increase in strength and decrease in ductility with increasing SA content. In vitro hydrolytic degradation studies indicated surface-eroding behavior. The degradation rate decreased with an increase of SA content in the polymers because of increased crystallinity. The release studies of both hydrophobic and hydrophilic dyes followed zero-order kinetics. In vitro cell studies to assess the cytotoxicity of the polymer confirmed minimal toxicity of the degradation products. Thus, a family of CO-SA polyanhydrides have been synthesized and characterized for controlled release applications where the physical, mechanical, and degradation kinetics can be modulated by varying the weight fraction of the prepolymers.

Effect of angle of incidence on mixed convective wake dynamics and heat transfer past a square cylinder in cross flow at Re=100

In this paper, a numerical investigation is performed to study the mixed convective flow and heat transfer characteristics past a square cylinder in cross flow at incidence. Utilizing air (Pr = 0.71) as an operating fluid, computations are carried out at a representative Reynolds number (Re) of 100. Angles of incidences are varied as, 0 degrees <= alpha <= 45 degrees. Effect of superimposed positive and negative cross-flow buoyancy is brought about by varying the Richardson number (RI) in the range -1.0 <= Ri <= 1.0. The detail features of flow topology and heat transport are analyzed critically for different angles of incidences. The thermo fluidic forces acting on the cylinder during mixed convection are captured in terms of the drag (C-D), lift (C-L), and moment (C-M) coefficients. The results show that the lateral width of the cylinder wake reduces with increasing alpha and the isotherms spread out far wide. In the range 0 degrees < alpha < 45 degrees, C-D reduces with increasing Ri. The functional dependence of C-M with Ri reveals a linear relationship. Thermal boundary layer thickness reduces with increasing angle of incidences. The global rate of heat transfer from the cylinder increases with increasing alpha. (C) 2014 Elsevier Ltd. All rights reserved.

Switching and Release Time Analysis of Electrostatically Actuated Capacitive RF MEMS Switches

This paper explains the reason behind pull-in time being more than pull-up time of many Radio Frequency Micro-Electro-Mechanical Systems (RF MEMS) switches at actuation voltages comparable to the pull-in voltage. Analytical expressions for pull-in and pull-up time are also presented. Experimental data as well as finite element simulations of electrostatically actuated beams used in RF-MEMS switches show that the pull-in time is generally more than the pull-up time. Pull-in time being more than pull-up time is somewhat counter-intuitive because there is a much larger electrostatic force during pull-in than the restoring mechanical force during the release. We investigated this issue analytically and numerically using a 1D model for various applied voltages and attribute this to energetics, the rate at which the forces change with time, and softening of the overall effective stiffness of the electromechanical system. 3D finite element analysis is also done to support the 1D model-based analyses.

Photonic Hydrogel Beads for Controlled Release of Risedronate

pH-sensitive photonic composite hydrogel beads composed of sodium alginate and risedronate sodium (SA/RIS) was prepared crosslinked by Ca2+ owing to the ionic gelation of SA. The structure and surface morphology of the composite hydrogel beads were characterized by SEM. pH-sensitivity of these composite hydrogels beads and the release behaviors of drug from them were investigated. The results showed that the composite hydrogel beads had good pH-sensitivity. The drug loading and encapsulation efficiency were 27.7% and 92% for RIS, respectively. The cumulative release ratios of RIS from the composite hydrogel beads were 2.47% in pH 2.1 solution and 83 % in pH 6.8 solutions within 24 h, respectively. However, the cumulative release ratio of RIS in pH 7.4 solution reached 91% within 7 h. It is proposed that the novel photonic SA/RIS composite hydrogel bead could possess the potential of an increased intestinal absorption and fewer adverse effects of RIS. The pH and salt response of photonic hydrogel bead, as well as the encapsulation of macromolecules, are promising for applications in biomedicine and biotechnology.

Role of surface chemistry in modulating drug release kinetics in titania nanotubes

Surface chemistry and the intrinsic porous architectures of porous substrates play a major role in the design of drug delivery systems. An interesting example is the drug elution characteristic from hydrothermally synthesised titania nanotubes with tunable surface chemistry. The variation in release rates of Ibuprofen (IBU) is largely influenced by the nature of the functional groups on titania nanotubes and pH of suspending medium. To elucidate the extent of interaction between the encapsulated IBU and the functional groups on titania nanotubes, the release profiles have been modelled with an empirical Hill equation. The analysis aided in establishing a probable mechanism for the release of IBU from the titania nanotubes. The study of controlled drug release from TiO2 has wider implication in the context of biomedical engineering. (C) 2014 Elsevier B.V. All rights reserved.

Photocatalytic disassembly of tertiary amine-based dendrimers to monomers and their application to the `catch and release' of a dye in aqueous solution

Photocatalytic disassembly of tertiary amine-based poly(propyl ether imine) dendrimers, in the presence of either 9,10-anthraquinone or riboflavin tetraacetate and O-2(g), leads to di- and tripropanolamine monomers. An application is shown by solubilisation of a water-insoluble dye, Sudan I, in aq. dendrimer solution ('catch'), followed by its `release' upon disassembly of the dendrimer.

MAXIMUM A-POSTERIORI ESTIMATION OF MISSING SAMPLES WITH CONTINUITY CONSTRAINT IN ELECTROMAGNETIC ARTICULOGRAPHY DATA

Electromagnetic Articulography (EMA) technique is used to record the kinematics of different articulators while one speaks. EMA data often contains missing segments due to sensor failure. In this work, we propose a maximum a-posteriori (MAP) estimation with continuity constraint to recover the missing samples in the articulatory trajectories recorded using EMA. In this approach, we combine the benefits of statistical MAP estimation as well as the temporal continuity of the articulatory trajectories. Experiments on articulatory corpus using different missing segment durations show that the proposed continuity constraint results in a 30% reduction in average root mean squared error in estimation over statistical estimation of missing segments without any continuity constraint.

Solving Dynamic Constraint Single Objective Functions using a Nature Inspired Technique

We present in this paper a new algorithm based on Particle Swarm Optimization (PSO) for solving Dynamic Single Objective Constrained Optimization (DCOP) problems. We have modified several different parameters of the original particle swarm optimization algorithm by introducing new types of particles for local search and to detect changes in the search space. The algorithm is tested with a known benchmark set and compare with the results with other contemporary works. We demonstrate the convergence properties by using convergence graphs and also the illustrate the changes in the current benchmark problems for more realistic correspondence to practical real world problems.

PECVD grown SiC cantilevers with Dry and Wet Release

Cantilevers made out of PECVD grown SiC films are reported here. The cantilevers were realized in two different methods isotropic etch (Dry release) and combination of wet etch and critical point dry release. The dry release process for Silicon isotropic etch results in excellent etch selectivity against SiC, to provide released structures. The optimized wet release process is able to overcome stiction issues to provide excellent SiC cantilevers.