Shaping Giant Membrane Vesicles in 3D-Printed Protein Hydrogel Cages

Giant unilamellar phospholipid vesicles are attractive starting points for constructing minimal living cells from the bottom-up. Their membranes are compatible with many physiologically functional modules and act as selective barriers, while retaining a high morphological flexibility. However, their spherical shape renders them rather inappropriate to study phenomena that are based on distinct cell shape and polarity, such as cell division. Here, a microscale device based on 3D printed protein hydrogel is introduced to induce pH-stimulated reversible shape changes in trapped vesicles without compromising their free-standing membranes. Deformations of spheres to at least twice their aspect ratio, but also toward unusual quadratic or triangular shapes can be accomplished. Mechanical force induced by the cages to phase-separated membrane vesicles can lead to spontaneous shape deformations, from the recurrent formation of dumbbells with curved necks between domains to full budding of membrane domains as separate vesicles. Moreover, shape-tunable vesicles are particularly desirable when reconstituting geometry-sensitive protein networks, such as reaction-diffusion systems. In particular, vesicle shape changes allow to switch between different modes of self-organized protein oscillations within, and thus, to influence reaction networks directly by external mechanical cues

TrustedCI: The NSF Cybersecurity Center of Excellence Globus Auth First Principles Vulnerability Assessment

The final report of the Globus Auth First Principles Vulnerability Assessment.NSF Grant # 1920430Ope

Real-scale investigation of the kinematic response of a rockfall protection embankment

This paper addresses the response of rockfall protection embankments when exposed to a rock impact. For this purpose, real-scale impact experiments were conducted with impact energies ranging from 200 to 2200 kJ. The structure was composed of a 4m high cellular wall leaned against a levee. The wall was a double-layer sandwich made from gabion cages filled with either stones or a sand–schreddedtyre mixture. For the first time, sensors were placed in different locations within the structure to measure real-time accelerations and displacements. The test conditions, measurement methods and results are presented in detail. The structure’s response is discussed in a descriptive and phenomenological approach and compared with previous real-scale experiments on other types of embankments

Geochemistry and cosmochemistry of fullerenes 3: Reaction of C60 and C70 with ozone

C60 and C70 dissolved in toluene were treated with O2 gas containing 2.6 volume percent ozone and with O3-free oxygen. No reaction products were detected for 0.1 mole of O2 passed through the solution, but destruction of C60 was clearly detectable for a dose of 10(exp -6) moles of O3. C70 was destroyed more slowly than C60. Among the substances remaining in solution, we identified C60O, C70O, C60O2, C60O3, and C60O4. C60 crystals exposed to O3 at room temperature became less soluble in toluene in a matter of days, but oxides were apparently not formed

Membrane-coated 3D architectures for bottom-up synthetic biology dagger

One of the great challenges of bottom-up synthetic biology is to recreate the cellular geometry and surface functionality required for biological reactions. Of particular interest are lipid membrane interfaces where many protein functions take place. However, cellular 3D geometries are often complex, and custom-shaping stable lipid membranes on relevant spatial scales in the micrometer range has been hard to accomplish reproducibly. Here, we use two-photon direct laser writing to 3D print microenvironments with length scales relevant to cellular processes and reactions. We formed lipid bilayers on the surfaces of these printed structures, and we evaluated multiple combinatorial scenarios, where physiologically relevant membrane compositions were generated on several different polymer surfaces. Functional dynamic protein systems were reconstituted in vitro and their self-organization was observed in response to the 3D geometry. This method proves very useful to template biological membranes with an additional spatial dimension, and thus allows a better understanding of protein function in relation to the complex morphology of cells and organelles.We also thank the Biochemistry Core Facility of the Max Planck Institute of Biochemistry for assistance with protein purification, and the Imaging Core Facility of the same institution for assistance on the 4D image visualisation

Vulnerability Assessment Enhancement for Middleware for Computing and Informatics

Security on Grid computing is often an afterthought. However assessing security of middleware systems is of the utmost importance because they manage critical resources owned by different organizations. To fulfill this objective we use First Principles Vulnerability Assessment (FPVA), an innovative analystic-centric (manual) methodology that goes beyond current automated vulnerability tools. FPVA involves several stages for characterizing the analyzed system and its components. Based on the evaluation of several middleware systems, we have found that there is a gap between the initial and the last stages of FPVA, which is filled with the security practitioner expertise. We claim that this expertise is likely to be systematically codified in order to be able to automatically indicate which, and why, components should be assessed. In this paper we introduce key elements of our approach: Vulnerability graphs, Vulnerability Graph Analyzer, and a Knowledge Base of security configurations

Unstable Attractors: Existence and Robustness in Networks of Oscillators With Delayed Pulse Coupling

We consider unstable attractors; Milnor attractors $A$ such that, for some neighbourhood $U$ of $A$, almost all initial conditions leave $U$. Previous research strongly suggests that unstable attractors exist and even occur robustly (i.e. for open sets of parameter values) in a system modelling biological phenomena, namely in globally coupled oscillators with delayed pulse interactions. In the first part of this paper we give a rigorous definition of unstable attractors for general dynamical systems. We classify unstable attractors into two types, depending on whether or not there is a neighbourhood of the attractor that intersects the basin in a set of positive measure. We give examples of both types of unstable attractor; these examples have non-invertible dynamics that collapse certain open sets onto stable manifolds of saddle orbits. In the second part we give the first rigorous demonstration of existence and robust occurrence of unstable attractors in a network of oscillators with delayed pulse coupling. Although such systems are technically hybrid systems of delay differential equations with discontinuous `firing' events, we show that their dynamics reduces to a finite dimensional hybrid system system after a finite time and hence we can discuss Milnor attractors for this reduced finite dimensional system. We prove that for an open set of phase resetting functions there are saddle periodic orbits that are unstable attractors.Comment: 29 pages, 8 figures,submitted to Nonlinearit