Manufacture of Multilayered Artificial Cell Membranes through Sequential Bilayer Deposition on Emulsion Templates

Efforts to manufacture artificial cells that replicate the architectures, processes and behaviours of biological cells are rapidly increasing. Perhaps the most commonly reconstructed cellular structure is the membrane, through the use of unilamellar vesicles as models. However, many cellular membranes, including bacterial double membranes, nuclear envelopes, and organelle membranes, are multilamellar. Due to a lack of technologies available for their controlled construction, multilayered membranes are not part of the repertoire of cell-mimetic motifs used in bottom-up synthetic biology. To address this, we developed emulsion-based technologies that allow cell-sized multilayered vesicles to be produced layer-by-layer, with compositional control over each layer, thus enabling studies that would otherwise remain inaccessible. We discovered that bending rigidities scale with the number of layers and demonstrate inter-bilayer registration between coexisting liquid–liquid domains. These technologies will contribute to the exploitation of multilayered membrane structures, paving the way for incorporating protein complexes that span multiple bilayers.     

Maternal N-Acetyl-Cysteine Prevents Neonatal Hypoxia-Induced Brain Injury in a Rat Model

Perinatal hypoxia is a major cause of infant brain damage, lifelong neurological disability, and infant mortality. N-Acetyl-Cysteine (NAC) is a powerful antioxidant that acts directly as a scavenger of free radicals. We hypothesized that maternal-antenatal and offspring-postnatal NAC can protect offspring brains from hypoxic brain damage.Sixty six newborn rats were randomized into four study groups. Group 1: Control (CON) received no hypoxic intervention. Group 2: Hypoxia (HYP)-received hypoxia protocol. Group 3: Hypoxia-NAC (HYP-NAC). received hypoxia protocol and treated with NAC following each hypoxia episode. Group 4: NAC Hypoxia (NAC-HYP) treated with NAC during pregnancy, pups subject to hypoxia protocol. Each group was evaluated for: neurological function (Righting reflex), serum proinflammatory IL-6 protein levels (ELISA), brain protein levels: NF-κB p65, neuronal nitric oxide synthase (nNOS), TNF-α, and IL-6 (Western blot) and neuronal apoptosis (histology evaluation with TUNEL stain). Hypoxia significantly increased pups brain protein levels compared to controls. NAC administration to dams or offspring demonstrated lower brain NF-κB p65, nNOS, TNF-α and IL-6 protein levels compared to hypoxia alone. Hypoxia significantly increased brain apoptosis as evidenced by higher grade of brain TUNEL reaction. NAC administration to dams or offspring significantly reduce this effect. Hypoxia induced acute sensorimotor dysfunction. NAC treatment to dams significantly attenuated hypoxia-induced acute sensorimotor dysfunction. Prophylactic NAC treatment of dams during pregnancy confers long-term protection to offspring with hypoxia associated brain injury, measured by several pathways of injury and correlated markers with pathology and behavior. This implies we may consider prophylactic NAC treatment for patients at risk for hypoxia during labor.     

Molecular Mechanisms Underlying TDP-43 Pathology in Cellular and Animal Models of ALS and FTLD

Amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD) are neurodegenerative disorders that exist on a disease spectrum due to pathological, clinical and genetic overlap. In up to 97% of ALS cases and ~50% of FTLD cases, the primary pathological protein observed in affected tissues is TDP-43, which is hyperphosphorylated, ubiquitinated and cleaved. The TDP-43 is observed in aggregates that are abnormally located in the cytoplasm. The pathogenicity of TDP-43 cytoplasmic aggregates may be linked with both a loss of nuclear function and a gain of toxic functions. The cellular processes involved in ALS and FTLD disease pathogenesis include changes to RNA splicing, abnormal stress granules, mitochondrial dysfunction, impairments to axonal transport and autophagy, abnormal neuromuscular junctions, endoplasmic reticulum stress and the subsequent induction of the unfolded protein response. Here, we review and discuss the evidence for alterations to these processes that have been reported in cellular and animal models of TDP-43 proteinopathy.     

The Prominin-1-Derived Peptide Improves Cardiac Function Following Ischemia

Myocardial infarction (MI) remains the leading cause of death in the western world. Despite advancements in interventional revascularization technologies, many patients are not candidates for them due to comorbidities or lack of local resources. Non-invasive approaches to accelerate revascularization within ischemic tissues through angiogenesis by providing Vascular Endothelial Growth Factor (VEGF) in protein or gene form has been effective in animal models but not in humans likely due to its short half-life and systemic toxicity. Here, we tested the hypothesis that PR1P, a small VEGF binding peptide that we developed, which stabilizes and upregulates endogenous VEGF, could be used to improve outcome from MI in rodents. To test this hypothesis, we induced MI in mice and rats via left coronary artery ligation and then treated animals with every other day intraperitoneal PR1P or scrambled peptide for 14 days. Hemodynamic monitoring and echocardiography in mice and echocardiography in rats at 14 days showed PR1P significantly improved multiple functional markers of heart function, including stroke volume and cardiac output. Furthermore, molecular biology and histological analyses of tissue samples showed that systemic PR1P targeted, stabilized and upregulated endogenous VEGF within ischemic myocardium. We conclude that PR1P is a potential non-invasive candidate therapeutic for MI.     

Novel algorithms for the network lifetime problem in wireless settings

One of the major concerns in wireless ad-hoc networks design is energy efficiency. Wireless devices are typically equipped with a limited energy supply sufficient only for a limited amount of time which is reversely proportional to the transmission power of the device. The network lifetime is defined as the time the first device runs out of its initial energy charge. In this paper we study the maximum network lifetime problem for broadcast and data gathering in wireless settings. We provide polynomial time approximation algorithms, with guaranteed performance bounds while considering omnidirectional and unidirectional transmissions. We also consider an extended variant of the maximum lifetime problem, which simultaneously satisfies additional constraints, such as bounded hop-diameter and degree of the routing tree, and minimizing the total energy used in a single transmission. Finally, we evaluate the performance of some of our algorithms through simulations.     

Delayed yielding of epoxy resin under tension,compression, and flexure. I. Behavior under constant strain rate

Series of loading tests were carried out on epoxy resin specimens, at varying constant strain rates, under tension, compression, and flexure. The stress–strain relationship revealed a distinct yielding stage followed shortly by a post-yielding region of decreasing load. In all cases, results indicate linearity between yield stress and log strain rate, in accordance with Eyring's theory of viscous flow. For specimens unloaded close to the yield point, photoelastic observations revealed a residual pattern parallel to the theoretical principal shear stresses. These results, supported by additional data from other works, indicate a viscoplastic deviatoric stress-biased diffusional mechanism as the dominant factor in the yielding of an amorphous crosslinked epoxy system.     

Is there more to complicated grief than depression and posttraumatic stress disorder? A test of incremental validity.

There is growing interest in complicated grief reactions as a possible new diagnostic category for inclusion in the Diagnostic and Statistical Manual of Mental Disorders. However, no research has yet shown that complicated grief has incremental validity (i.e., predicts unique variance in functioning). The authors addressed this issue in 2 studies by comparing grief, depression, and posttraumatic stress disorder (PTSD) symptoms with different measures of functioning (interviewer ratings, friend ratings, self-report, and autonomic arousal). The 1st study (N = 73) used longitudinal data collected at 4 and 18 months postloss, and the 2nd study (N = 447) used cross-sectional data collected 2.5-3.5 years postloss. With depression and PTSD controlled, grief emerged as a unique predictor of functioning, both cross-sectionally and prospectively. The findings provide convergent support for the incremental validity of complicated grief as an independent marker of bereavement-related psychopathology. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Modeling the Heat Wave Generated during Microwave Heating of Powdered Zinc Oxide in a Nitrogen Atmosphere

A theoretical investigation of previous experiments on the microwave heating of cylindrical samples of powdered ZnO in nitrogen and air environments has been presented. Unlike heating in air, which showed an exponentially saturating temperature behavior, a large discontinuity in the measured temperature was observed in nitrogen. Energy considerations suggested that the microwave power was mostly absorbed in a narrow, localized region that moved radially outward as a propagating heat wave. It was assumed that this behavior was caused by an anomaly in the temperature dependence of the permittivity, and the present simulations verify this assumption. The phenomenon has been investigated using a newly developed two-dimensional code, and the results are in good agreement with the experimental observations. The calculated width of the microwave absorbing region was ∼1 mm, and the radial velocity of the electromagnetically driven heat wave was inversely proportional to the square of the distance from the center of the sample, with a maximum speed of ∼20 mm/min.     

CoRAL: A transparent fault-tolerant web service

The Web is increasingly used for critical applications and services. We present a client-transparent mechanism, called CoRAL, that provides high reliability and availability for Web service. CoRAL provides fault tolerance even for requests being processed at the time of server failure. The scheme does not require deterministic servers and can thus handle dynamic content. CoRAL actively replicates the TCP connection state while maintaining logs of HTTP requests and replies. In the event of a primary server failure, active client connections fail over to a spare, where their processing continues seamlessly. We describe key aspects of the design and implementation as well as several performance optimizations. Measurements of system overhead, failover performance, and preliminary validation using fault injection are presented.