Plasmodium falciparum glutamate dehydrogenase a is dispensable and not a drug target during erythrocytic development

<p>Background: Plasmodium falciparum contains three genes encoding potential glutamate dehydrogenases. The protein encoded by gdha has previously been biochemically and structurally characterized. It was suggested that it is important for the supply of reducing equivalents during intra-erythrocytic development of Plasmodium and, therefore, a suitable drug target.</p> <p>Methods: The gene encoding the NADP(H)-dependent GDHa has been disrupted by reverse genetics in P. falciparum and the effect on the antioxidant and metabolic capacities of the resulting mutant parasites was investigated.</p> <p>Results: No growth defect under low and elevated oxygen tension, no up-or down-regulation of a number of antioxidant and NADP(H)-generating proteins or mRNAs and no increased levels of GSH were detected in the D10(Delta gdha) parasite lines. Further, the fate of the carbon skeleton of [(13)C] labelled glutamine was assessed by metabolomic studies, revealing no differences in the labelling of a-ketoglutarate and other TCA pathway intermediates between wild type and mutant parasites.</p> <p>Conclusions: First, the data support the conclusion that D10(Delta gdha) parasites are not experiencing enhanced oxidative stress and that GDHa function may not be the provision of NADP(H) for reductive reactions. Second, the results imply that the cytosolic, NADP(H)-dependent GDHa protein is not involved in the oxidative deamination of glutamate but that the protein may play a role in ammonia assimilation as has been described for other NADP(H)dependent GDH from plants and fungi. The lack of an obvious phenotype in the absence of GDHa may point to a regulatory role of the protein providing glutamate (as nitrogen storage molecule) in situations where the parasites experience a limiting supply of carbon sources and, therefore, under in vitro conditions the enzyme is unlikely to be of significant importance. The data imply that the protein is not a suitable target for future drug development against intra-erythrocytic parasite development.</p&gt

High-Level Information Fusion in Visual Sensor Networks

Information fusion techniques combine data from multiple sensors, along with additional information and knowledge, to obtain better estimates of the observed scenario than could be achieved by the use of single sensors or information sources alone. According to the JDL fusion process model, high-level information fusion is concerned with the computation of a scene representation in terms of abstract entities such as activities and threats, as well as estimating the relationships among these entities. Recent experiences confirm that context knowledge plays a key role in the new-generation high-level fusion systems, especially in those involving complex scenarios that cause the failure of classical statistical techniques –as it happens in visual sensor networks. In this chapter, we study the architectural and functional issues of applying context information to improve high-level fusion procedures, with a particular focus on visual data applications. The use of formal knowledge representations (e.g. ontologies) is a promising advance in this direction, but there are still some unresolved questions that must be more extensively researched.The UC3M Team gratefully acknowledges that this research activity is supported in part by Projects CICYT TIN2008-06742-C02-02/TSI, CICYT TEC2008-06732-C02-02/TEC, CAM CONTEXTS (S2009/TIC-1485) and DPS2008-07029-C02-02

Research Opportunities in Contextualized Fusion Systems. The Harbor Surveillance Case

Proceedings of: International Workshop of Intelligent Systems for Context-Based Information Fusion (ISCIF 2011) associated to 11th International Work-Conference on Artificial Neural Networks, IWANN, Torremolinos-Málaga, Spain, June 8-10, 2011.The design of modern Information Fusion (IF) systems involves a complex process to achieve the requirements in the selected applications, especially in domains with a high degree of customization. In general, an advanced fusion system is required to show robust, context-sensitive behavior and efficient performance in real time. It is necessary to exploit all potentially relevant sensor and contextual information in the most appropriate way. Among modern applications for IF technology is the case of surveillance of complex harbor environments that are comprised of large numbers of surface vessels, high-value and dangerous facilities, and many people. The particular conditions and open needs in the harbor scenario are reviewed in this paper, highlighting research opportunities to explore in the development of fusion systems in this area.This work was supported in part by Projects CICYT TIN2008-06742-C02-02/TSI, CICYT TEC2008-06732-C02-02/TEC and CAM CONTEXTS S2009/TIC-1485.Publicad

A role for fast rhythmic bursting neurons in cortical gamma oscillations in vitro

Basic cellular and network mechanisms underlying gamma frequency oscillations (30–80 Hz) have been well characterized in the hippocampus and associated structures. In these regions, gamma rhythms are seen as an emergent property of networks of principal cells and fast-spiking interneurons. In contrast, in the neocortex a number of elegant studies have shown that specific types of principal neuron exist that are capable of generating powerful gamma frequency outputs on the basis of their intrinsic conductances alone. These fast rhythmic bursting (FRB) neurons (sometimes referred to as "chattering" cells) are activated by sensory stimuli and generate multiple action potentials per gamma period. Here, we demonstrate that FRB neurons may function by providing a large-scale input to an axon plexus consisting of gap-junctionally connected axons from both FRB neurons and their anatomically similar counterparts regular spiking neurons. The resulting network gamma oscillation shares all of the properties of gamma oscillations generated in the hippocampus but with the additional critical dependence on multiple spiking in FRB cells

PNAS plus: plasmodium falciparum responds to amino acid starvation by entering into a hibernatory state

The human malaria parasite Plasmodium falciparum is auxotrophic for most amino acids. Its amino acid needs are met largely through the degradation of host erythrocyte hemoglobin; however the parasite must acquire isoleucine exogenously, because this amino acid is not present in adult human hemoglobin. We report that when isoleucine is withdrawn from the culture medium of intraerythrocytic P. falciparum, the parasite slows its metabolism and progresses through its developmental cycle at a reduced rate. Isoleucine-starved parasites remain viable for 72 h and resume rapid growth upon resupplementation. Protein degradation during starvation is important for maintenance of this hibernatory state. Microarray analysis of starved parasites revealed a 60% decrease in the rate of progression through the normal transcriptional program but no other apparent stress response. Plasmodium parasites do not possess a TOR nutrient-sensing pathway and have only a rudimentary amino acid starvation-sensing eukaryotic initiation factor 2α (eIF2α) stress response. Isoleucine deprivation results in GCN2-mediated phosphorylation of eIF2α, but kinase-knockout clones still are able to hibernate and recover, indicating that this pathway does not directly promote survival during isoleucine starvation. We conclude that P. falciparum, in the absence of canonical eukaryotic nutrient stress-response pathways, can cope with an inconsistent bloodstream amino acid supply by hibernating and waiting for more nutrient to be provided

Stimulus - response curves of a neuronal model for noisy subthreshold oscillations and related spike generation

We investigate the stimulus-dependent tuning properties of a noisy ionic conductance model for intrinsic subthreshold oscillations in membrane potential and associated spike generation. On depolarization by an applied current, the model exhibits subthreshold oscillatory activity with occasional spike generation when oscillations reach the spike threshold. We consider how the amount of applied current, the noise intensity, variation of maximum conductance values and scaling to different temperature ranges alter the responses of the model with respect to voltage traces, interspike intervals and their statistics and the mean spike frequency curves. We demonstrate that subthreshold oscillatory neurons in the presence of noise can sensitively and also selectively be tuned by stimulus-dependent variation of model parameters.Comment: 19 pages, 7 figure

Strategies and Techniques for Use and Exploitation of Contextual Information in High-Level Fusion Architectures

Proceedings of: 13th Conference on Information Fusion (FUSION 2010): Edinburgh, UK. 26-29 July 2010.Contextual Information is proving to be not only an additional exploitable information source for improving entity and situational estimates in certain Information Fusion systems, but can also be the entire focus of estimation for such systems as those directed to Ambient Intelligence (AI) and Context-Aware(CA) applications. This paper will discuss the role(s) of Contextual Information (CI) in a wide variety of IF applications to include AI, CA, Defense, and Cyber-security among possible others, the issues involved in designing strategies and techniques for CI use and exploitation, provide some exemplars of evolving CI use/exploitation designs on our current projects, and describe some general frameworks that are evolving in various application domains where CI is proving critical.The UC3M Team gratefully acknowledge that this research activity is supported in part by Projects CICYT TIN2008-06742-C02-02/TSI, CICYT TEC2008-06732- C02-02/TEC, CAM CONTEXTS (S2009/TIC-1485) and DPS2008-07029-C02-02. UC3M also thanks Prof. James Llinas for his helpful comments during his stay, which has been supported by the collaboration agreement ‘Chairs of Excellence’ between University Carlos III and Banco Santander. The US/UB Team gratefully acknowledge that this research activity is supported by a Multidisciplinary University Research Initiative (MURI) grant (Number W911NF-09-1-0392) for “Unified Research on Networkbased Hard/Soft Information Fusion”, issued by the US Army Research Office (ARO) under the program management of Dr. John LaveryPublicad

3',5'-Cyclic Adenosine Monophosphate- and Ca2+-Calmodulin-Dependent Endogenous Protein Phosphorylation Activity in Membranes of the Bovine Chromaffin Secretory Vesicles: Identification of Two Phosphorylated Components as Tyrosine Hydroxylase and Protein Kinase Regulatory Subunit Type II

Abstract: Membranes of the secretory vesicles from bovine adrenal medulla were investigated for the presence of the endogenous protein phosphorylation activity. Seven phosphoprotein bands in the molecular weight range of 250,000 to 30,000 were observed by means of the sodium dodecyl sulphate electrophoresis and autoradiography. On the basis of the criteria of molecular weight, selective stimulation of the phosphorylation by cyclic AMP (as compared with cyclic GMP) and immunoprecipitation by specific antibodies, band 5 (molecular weight 60,300) was found to represent the phosphorylated form of the secretory vesicle-bound tyrosine hydroxylase. The electrophoretic mobility, the stimulatory and inhibitory effects of cyclic AMP in presence of Mg2+ and Zn,2+ respectively, and immunoreactivity toward antibodies showed band 6 to contain two forms of the regulatory subunits of the type II cyclic AMP-dependent protein kinase, distinguishable by their molecular weights (56,000 and 52,000, respectively). Phosphorylation of band 7 (molecular weight 29,800) was stimulated about 2 to 3 times by Ca2+ and calmodulin in the concentration range of both agents believed to occur in the secretory tissues under physiological conditions

The Case for Dynamic Models of Learners' Ontologies in Physics

In a series of well-known papers, Chi and Slotta (Chi, 1992; Chi & Slotta, 1993; Chi, Slotta & de Leeuw, 1994; Slotta, Chi & Joram, 1995; Chi, 2005; Slotta & Chi, 2006) have contended that a reason for students' difficulties in learning physics is that they think about concepts as things rather than as processes, and that there is a significant barrier between these two ontological categories. We contest this view, arguing that expert and novice reasoning often and productively traverses ontological categories. We cite examples from everyday, classroom, and professional contexts to illustrate this. We agree with Chi and Slotta that instruction should attend to learners' ontologies; but we find these ontologies are better understood as dynamic and context-dependent, rather than as static constraints. To promote one ontological description in physics instruction, as suggested by Slotta and Chi, could undermine novices' access to productive cognitive resources they bring to their studies and inhibit their transition to the dynamic ontological flexibility required of experts.Comment: The Journal of the Learning Sciences (In Press