Vapor nanobubble is the more reliable photothermal mechanism for inducing endosomal escape of siRNA without disturbing cell homeostasis

Strategies for controlled delivery of therapeutic siRNA into living cells are in high demand as endosomal escape remains the most prominent bottleneck at the intracellular level. Photothermal properties of gold nanoparticles (AuNP) can be used to overcome the endosomal membrane barrier upon laser irradiation by two mechanisms: endosomal rupture by mechanical energy from water vapor nanobubbles (VNBs), or permeabilization of the endosomal membrane by heat diffusion. Here we evaluated how both mechanisms influence cargo release, transfection efficiency, acute cytotoxicity and cell homeostasis. Using a siRNA/AuNP drug delivery system we found that the in vitro release of siRNA from the AuNP carrier occurs equally efficiently by VNB formation or heat generation. Heat-mediated endosomal escape happened more efficiently in cells that had more particles per endosome, resulting in variable siRNA-induced downregulation (20-50%). VNB-mediated endosomal escape did not dependent on the number of AuNP per endosome, yielding high downregulations (50-60%) independent of the cell type. Effects on cell homeostasis by whole transcriptome analysis, showed a quick recover after 24 h or 48 h for either of both photothermal mechanisms. We conclude that VNBs are more consistent to induce efficient endosomal escape and gene silencing independent of the cell type without long lasting effects on cell homeostasis

Modelling and in vitro testing of the HIV-1 Nef fitness landscape.

An effective vaccine is urgently required to curb the HIV-1 epidemic. We have previously described an approach to model the fitness landscape of several HIV-1 proteins, and have validated the results against experimental and clinical data. The fitness landscape may be used to identify mutation patterns harmful to virus viability, and consequently inform the design of immunogens that can target such regions for immunological control. Here we apply such an analysis and complementary experiments to HIV-1 Nef, a multifunctional protein which plays a key role in HIV-1 pathogenesis. We measured Nef-driven replication capacities as well as Nef-mediated CD4 and HLA-I down-modulation capacities of thirty-two different Nef mutants, and tested model predictions against these results. Furthermore, we evaluated the models using 448 patient-derived Nef sequences for which several Nef activities were previously measured. Model predictions correlated significantly with Nef-driven replication and CD4 down-modulation capacities, but not HLA-I down-modulation capacities, of the various Nef mutants. Similarly, in our analysis of patient-derived Nef sequences, CD4 down-modulation capacity correlated the most significantly with model predictions, suggesting that of the tested Nef functions, this is the most important in vivo. Overall, our results highlight how the fitness landscape inferred from patient-derived sequences captures, at least in part, the in vivo functional effects of mutations to Nef. However, the correlation between predictions of the fitness landscape and measured parameters of Nef function is not as accurate as the correlation observed in past studies for other proteins. This may be because of the additional complexity associated with inferring the cost of mutations on the diverse functions of Nef

Distributed allocation of mobile sensing swarms in gyre flows

We address the synthesis of distributed control policies to enable a swarm of homogeneous mobile sensors to maintain a desired spatial distribution in a geophysical flow environment, or workspace. In this article, we assume the mobile sensors (or robots) have a "map" of the environment denoting the locations of the Lagrangian coherent structures or LCS boundaries. Based on this information, we design agent-level hybrid control policies that leverage the surrounding fluid dynamics and inherent environmental noise to enable the team to maintain a desired distribution in the workspace. We establish the stability properties of the ensemble dynamics of the distributed control policies. Since realistic quasi-geostrophic ocean models predict double-gyre flow solutions, we use a wind-driven multi-gyre flow model to verify the feasibility of the proposed distributed control strategy and compare the proposed control strategy with a baseline deterministic allocation strategy. Lastly, we validate the control strategy using actual flow data obtained by our coherent structure experimental testbed.Comment: 10 pages, 14 Figures, added reference

Effect of short time captopril administration on spatial memory in aging rats

Introduction: The brain renin-angiotensin system (RAS) has been reported having a pathological role in age-related impairment in learning and memory. Therefore, angiotensin converting enzyme inhibitors (ACEi) are expected to have positive ef fects on memory. Longtime treatment with captopril (an angiotensin converting enzyme inhibitor) significantly attenuates the age-related impairment in learning and memory. Materials and Methods: In the present study, 24 month old male Wistar rats were divided into four experimental groups (n=8). Captopril treated groups received daily ip injections of captopril at doses of 5, 10, 15 mg/kg/day for one week, the forth group served as control and remained untreated. Learning process was assessed by the reference memory task in the Morris water maze. All rats received water maze training (4 trials/day for 5 days) to assess hippocampal dependent spatial learning and then received a 60-s probe test of spatial memory retention 24 h after the 20th trial. Results: Over 5 days of training, captopril 5, 10, 15 mg/kg/day treatment signi ficantly reduced the latency and path length to finding the escape platform. In probe trails (without platform), on the last day of training, the captopril -treated group spent significantly longer time in the platform quadrant than control animals. Among treated group, 10 /mg/Kg dosage of captopril induced the best rehearsals memory. Conclusion: These results confirm the previous studies that ACEi have a positive influence on memory and it was noticeable that even short time treatment by captopril can improve spatial memory in the aged rats. © 2015, Iranian Society of Physiology and Pharmacology. All rights reserved

Transport of inertial particles by Lagrangian coherent structures : application to predator-prey interaction in jellyfish feeding

We use a dynamical systems approach to identify coherent structures from often chaotic motions of inertial particles in open flows. We show that particle Lagrangian coherent structures (pLCS) act as boundaries between regions in which particles have different kinematics. They provide direct geometric information about the motion of ensembles of inertial particles, which is helpful to understand their transport. As an application, we apply the methodology to a planktonic predator–prey system in which moon jellyfish Aurelia aurita uses its body motion to generate a flow that transports small plankton such as copepods to its vicinity for feeding. With the flow field generated by the jellyfish measured experimentally and the dynamics of plankton described by a modified Maxey–Riley equation, we use the pLCS to identify a capture region in which prey can be captured by the jellyfish. The properties of the pLCS and the capture region enable analysis of the effect of several physiological and mechanical parameters on the predator–prey interaction, such as prey size, escape force, predator perception, etc. The methods developed here are equally applicable to multiphase and granular flows, and can be generalized to any other particle equation of motion, e.g. equations governing the motion of reacting particles or charged particles

Efficacy of a sensory deterrent and pipe modifications in decreasing entrainment of juvenile green sturgeon (Acipenser medirostris) at unscreened water diversions.

Water projects designed to extract fresh water for local urban, industrial and agricultural use throughout rivers and estuaries worldwide have contributed to the fragmentation and degradation of suitable habitat for native fishes. The number of water diversions located throughout the Sacramento-San Joaquin watershed in California's Central Valley exceeds 3300, and the majority of these are unscreened. Many anadromous fish species are susceptible to entrainment into these diversions, potentially impacting population numbers. In the laboratory, juvenile green sturgeon (Acipenser medirostris) have been shown to have high entrainment rates into unscreened diversions compared with those of other native California fish species, which may act as a significant source of mortality for this already-threatened species. Therefore, we tested the efficacy of a sensory deterrent (strobe light) and two structural pipe modifications (terminal pipe plate and upturned pipe configuration) in decreasing the entrainment of juvenile green sturgeon (mean mass ± SEM = 162.9 ± 4.0 g; mean fork length = 39.4 ± 0.3 cm) in a large (>500 kl) outdoor flume fitted with a water-diversion pipe 0.46 m in diameter. While the presence of the strobe light did not affect fish entrainment rates, the terminal pipe plate and upturned pipe modifications significantly decreased the proportion of fish entrained out of the total number tested relative to control conditions (0.13 ± 0.02 and 0.03 ± 0.02 vs. 0.44 ± 0.04, respectively). These data suggest that sensory deterrents using visual stimuli are not an effective means to reduce diversion pipe interactions for green sturgeon, but that structural alterations to diversions can successfully reduce entrainment for this species. Our results are informative for the development of effective management strategies to mitigate the impacts of water diversions on sturgeon populations and suggest that effective restoration strategies that balance agricultural needs with conservation programmes are possible

Ultra-fast escape maneuver of an octopus-inspired robot

We design and test an octopus-inspired flexible hull robot that demonstrates outstanding fast-starting performance. The robot is hyper-inflated with water, and then rapidly deflates to expel the fluid so as to power the escape maneuver. Using this robot we verify for the first time in laboratory testing that rapid size-change can substantially reduce separation in bluff bodies traveling several body lengths, and recover fluid energy which can be employed to improve the propulsive performance. The robot is found to experience speeds over ten body lengths per second, exceeding that of a similarly propelled optimally streamlined rigid rocket. The peak net thrust force on the robot is more than 2.6 times that on an optimal rigid body performing the same maneuver, experimentally demonstrating large energy recovery and enabling acceleration greater than 14 body lengths per second squared. Finally, over 53% of the available energy is converted into payload kinetic energy, a performance that exceeds the estimated energy conversion efficiency of fast-starting fish. The Reynolds number based on final speed and robot length is $Re \approx 700,000$. We use the experimental data to establish a fundamental deflation scaling parameter $\sigma^*$ which characterizes the mechanisms of flow control via shape change. Based on this scaling parameter, we find that the fast-starting performance improves with increasing size.Comment: Submitted July 10th to Bioinspiration & Biomimetic