Phospholipase A2 activity and calpactin I levels in rat lymphokine-activated killer cells: correlation with the cytotoxic activity

The interaction of carnitine with human placental brush-border membrane vesicles was investigated. Carnitine was found to associate with the membrane vesicles in a Na(+)-dependent manner. The time course of this association did not exhibit an overshoot, which is typical of a Na+ gradient-driven transport process. The absolute requirement for Na+ was noticeable whether the association of carnitine with the vesicles was measured with a short time incubation or under equilibrium conditions, indicating Na(+)-dependent binding of carnitine to the human placental brush-border membranes. The binding was saturable and was of a high-affinity type with a dissociation constant of 1.37 +/- 0.03 microM. Anions had little or no influence on the binding process. The binding process was specific for carnitine and its acyl derivatives. Betaine also competed for the binding process, but other structurally related compounds did not. Kinetic analyses revealed that Na+ increased the affinity of the binding process for carnitine and the Na+/carnitine coupling ratio for the binding process was 1. The dissociation constant for the interaction of Na+ with the binding of carnitine was 24 +/- 4 mM. This constitutes the first report on the identification of Na(+)-dependent high-affinity carnitine binding in the plasma membrane of a mammalian cell. Studies with purified rat renal brush-border membrane vesicles demonstrated the presence of Na+ gradient-driven carnitine transport but no Na(+)-dependent carnitine binding in these membrane vesicles. In contrast, purified intestinal brush-border membrane vesicles posses neither Na+ gradient-driven carnitine transport nor Na(+)-dependent carnitine binding.     

Aerobic degradation by white‐rot fungi of trichloroethylene (TCE) and mixtures of TCE and perchloroethylene (PCE)

BACKGROUND: Trichloroethylene (TCE) and perchloroethylene (PCE) are considered among the most important groundwater pollutants around the world. These compounds are usually found together in polluted environments but little is known about the ability of microorganisms to simultaneously degrade TCE and PCE. RESULTS: Data showed that several species of white‐rot fungi, including Trametes versicolor, Ganoderma lucidum, and Irpex lacteus, degrade substantial levels of TCE in pure culture. T. versicolor was chosen for further study since it degraded higher levels of TCE than the other organisms. Initial glucose concentration and reoxygenation of samples increased the amount of TCE dechlorination, but no significant difference in percentage TCE degradation was observed. T. versicolor was able to degrade 34.1 and 47.7% of PCE and TCE added as mixtures (containing 5 and 10 mg L⁻¹, respectively). CONCLUSIONS: The degradation ability of TCE was extended to other species of white‐rot fungi. Percentage degradation as well as chloride release from mixtures of TCE and PCE showed that T. versicolor degrades mixtures of TCE and PCE almost as well as its ability to degrade individually added TCE or PCE. The results suggest the potential promise of T. versicolor for bioremediation of TCE and PCE in the environment. Copyright © 2008 Society of Chemical Industry     

External and internal electrostatic potentials of cholinesterase models

The electrostatic potentials for the three-dimensional structures of cholinesterases from various species were calculated, using the Delphi algorithm, on the basis of the Poisson–Boltzmann equation. We used structures for Torpedo californica and mouse acetylcholinesterase, and built homology models of the human, Bungarus fasciatus, and Drosophila melanogaster acetylcholinesterases and human butyrylcholinesterase. All these structures reveal a negative external surface potential, in the area around the entrance to the active-site gorge, that becomes more negative as the rim of the gorge is approached. Moreover, in all cases, the potential becomes increasingly more negative along the central axis running down the gorge, and is largest at the base of the gorge, near the active site. Ten key acidic residues conserved in the sequence alignments of AChE from various species, both in the surface area near the entrance of the active-site gorge and at its base, appear to be primarily responsible for these potentials. The potentials are highly correlated among the structures examined, down to sequence identities as low as 35%. This indicates that they are a conserved property of the cholinesterase family, could serve to attract the positively charged substrate into and down the gorge to the active site, and may play other roles important for cholinesterase function.     

Human-Induced Neural and Mesenchymal Stem Cell Therapy Combined with a Curcumin Nanoconjugate as a Spinal Cord Injury Treatment

We currently lack effective treatments for the devastating loss of neural function associated with spinal cord injury (SCI). In this study, we evaluated a combination therapy comprising human neural stem cells derived from induced pluripotent stem cells (iPSC-NSC), human mesenchymal stem cells (MSC), and a pH-responsive polyacetal–curcumin nanoconjugate (PA-C) that allows the sustained release of curcumin. In vitro analysis demonstrated that PA-C treatment protected iPSC-NSC from oxidative damage in vitro, while MSC co-culture prevented lipopolysaccharide-induced activation of nuclear factor-κB (NF-κB) in iPSC-NSC. Then, we evaluated the combination of PA-C delivery into the intrathecal space in a rat model of contusive SCI with stem cell transplantation. While we failed to observe significant improvements in locomotor function (BBB scale) in treated animals, histological analysis revealed that PA-C-treated or PA-C and iPSC-NSC + MSC-treated animals displayed significantly smaller scars, while PA-C and iPSC-NSC + MSC treatment induced the preservation of β-III Tubulin-positive axons. iPSC-NSC + MSC transplantation fostered the preservation of motoneurons and myelinated tracts, while PA-C treatment polarized microglia into an anti-inflammatory phenotype. Overall, the combination of stem cell transplantation and PA-C treatment confers higher neuroprotective effects compared to individual treatments.     

Enhancing decentralized service discovery in open service-oriented multi-agent systems

Service-oriented multi-agent systems are dynamic systems that are populated by heterogeneous agents. These agents model their functionality as services in order to allow heterogeneous agents or other entities to interact with each other in a standardized way. Furthermore, due to the large-scale and adaptative needs of the system, traditional directory facilitators or middle-agents are not suitable for the management of agent services. This article proposes the introduction of homophily in service-oriented multi-agent systems to create efficient decentralized and self-organized structures where agents have a greater probability of establishing links with similar agents than with dissimilar ones. This similarity is based on two social dimensions: the set of services that an agent provides and the organizational roles that it plays. A second contribution is an algorithm for service discovery that it is carried out taking into account the local information that is related to the homophily between agents. The experiments compare our proposal with other proposals in distributed environments. The results show that the proposed structure and algorithm offer desirable features for service discovery in decentralized environments. Specifically, these features provide short paths and a high success rate in the service discovery process and resilience under deliberate failures.     

Self-managed topologies in P2P networks

The problem of efficient resource location is an important open issue in P2P systems. This paper introduces DANTE, a self-adapting P2P system that changes its peer links to form topologies where resources are located in an efficient manner via random walks. Additionally, this same self-adaptation capacity makes DANTE capable of reacting to events like changes in the system load or attacks on well-connected nodes by adjusting the topology to the new scenario. This adaptive behavior emerges as the global result of the individual work of nodes, without the intervention of any central entity or the need for global knowledge. Simulations show that this adaptation process makes the system scalable, resilient to attacks, and tolerant to a high transitivity of peers. Simulations are also used to compare this solution with other well-known self-adapting P2P system. From these results it can be concluded that the topologies achieved by DANTE offer better performance.     

Interaction of miR-155 with Human Serum Albumin: An Atomic Force Spectroscopy,Fluorescence, FRET,and Computational Modelling Evidence

This study investigated the interaction between Human Serum Albumin (HSA) and microRNA 155 (miR-155) through spectroscopic, nanoscopic and computational methods. Atomic force spectroscopy together with static and time-resolved fluorescence demonstrated the formation of an HSA/miR-155 complex characterized by a moderate affinity constant (K_A in the order of 10⁴ M⁻¹). Förster Resonance Energy Transfer (FRET) experiments allowed us to measure a distance of (3.9 ± 0.2) nm between the lone HSA Trp214 and an acceptor dye bound to miR-155 within such a complex. This structural parameter, combined with computational docking and binding free energy calculations, led us to identify two possible models for the structure of the complex, both characterized by a topography in which miR-155 is located within two positively charged pockets of HSA. These results align with the interaction found for HSA and miR-4749, reinforcing the thesis that native HSA is a suitable miRNA carrier under physiological conditions for delivering to appropriate targets.     

Reversible Photocontrol of Dopaminergic Transmission in Wild-Type Animals

Understanding the dopaminergic system is a priority in neurobiology and neuropharmacology. Dopamine receptors are involved in the modulation of fundamental physiological functions, and dysregulation of dopaminergic transmission is associated with major neurological disorders. However, the available tools to dissect the endogenous dopaminergic circuits have limited specificity, reversibility, resolution, or require genetic manipulation. Here, we introduce azodopa, a novel photoswitchable ligand that enables reversible spatiotemporal control of dopaminergic transmission. We demonstrate that azodopa activates D₁-like receptors in vitro in a light-dependent manner. Moreover, it enables reversibly photocontrolling zebrafish motility on a timescale of seconds and allows separating the retinal component of dopaminergic neurotransmission. Azodopa increases the overall neural activity in the cortex of anesthetized mice and displays illumination-dependent activity in individual cells. Azodopa is the first photoswitchable dopamine agonist with demonstrated efficacy in wild-type animals and opens the way to remotely controlling dopaminergic neurotransmission for fundamental and therapeutic purposes.     

Superconducting Vortex Lattice Configurations on Periodic Potentials: Simulation and Experiment

Nb films have been fabricated on top of array of Ni nanodots. The array of periodic pinning potentials modifies the vortex lattice for specific values of the external applied magnetic field. By means of an implemented code developed from scratch, computer simulations based only on the vortex?Cvortex and the vortex?Cnanodot interactions provide the total interaction between vortices and pinning sites as well as the position of the vortices in the array unit cell. This simulation approach could be performed on square, rectangular or triangular arrays of nanodefects of different size.