Unveiling the components of the signaling pathway between endosomes and the phagocytic uptake machinery in Dictyostelium discoideum

The soil amoebae Dictyostelium discoideum take up particles from their environment in order to obtain nutrition. The particle transits through the cell within a phagosome that fuses with organelles of different molecular compositions, undergoing a gradual degradation by different sets of hydrolytic enzymes. Griffiths’ concept of “phagosome individuality” predicts signaling from phagosomes into the cytoplasm, which might regulate many aspects of cell physiology. The finding that Dictyostelium cells depleted of the lysozyme AlyA or over-expressing the esterase Gp70 exhibit increased uptake of food particles, led to the postulation of a signaling cascade between endocytic compartments and the cytoskeletal uptake machinery at the plasma membrane. Assuming that Gp70 acts downstream of AlyA, gene-expression profiling of both mutants revealed different and overlapping sets of misregulated genes that might participate in this signaling cascade. Based on these results, we analyzed the effects of the artificial misregulation of six candidate genes by over-expression or negative genetic interference, in order to reconstruct at least part of the signaling pathway. SSB420 and SSL793 were chosen as candidates for the first signaling step, as they were up-regulated in AlyA-null cells and remained unaltered in the Gp70 over-expressing cells. The over-expression of SSB420 enhanced phagocytosis and raised the expression levels of Gp70, supporting its involvement in the signaling pathway between AlyA and Gp70 as a positive regulator of phagocytosis. However, this was not the case of cells over-expressing SSL793, as this mutation had no effects on phagocytosis. For the signaling downstream of Gp70, we studied four commonly misregulated genes in AlyA-depleted and Gp70 over-expressing cells. The expression levels of SLB350, SSB389 and TipD were lower in both mutants and therefore these were assumed as possible candidates for the negative regulation of phagocytosis. Cells depleted of SLB350 exhibited an increased phagocytic activity and no effect on Gp70 expression, proving its participation in the signaling pathway downstream of Gp70. Unlike SLB350, the disruption of the genes coding for SSB389 and TipD had no effects on particle uptake, excluding them from the pathway. The fourth candidate was Yipf1, the only gene that was commonly up-regulated in both mutants. Yet, the artificial over-expression of this protein had no effects on phagocytosis, so this candidate is also not included in the signaling pathway. Furthermore, localizing the products of the candidate genes within the cell helped unveiling several cellular organelles that receive signals from the phagosome and transduce them towards the uptake machinery.

Targeting the cell cycle machinery for the treatment of cardiovascular disease

Cardiovascular disease represents a major clinical problem affecting a significant proportion of the world's population and remains the main cause of death in the UK. The majority of therapies currently available for the treatment of cardiovascular disease do not cure the problem but merely treat the symptoms. Furthermore, many cardioactive drugs have serious side effects and have narrow therapeutic windows that can limit their usefulness in the clinic. Thus, the development of more selective and highly effective therapeutic strategies that could cure specific cardiovascular diseases would be of enormous benefit both to the patient and to those countries where healthcare systems are responsible for an increasing number of patients. In this review, we discuss the evidence that suggests that targeting the cell cycle machinery in cardiovascular cells provides a novel strategy for the treatment of certain cardiovascular diseases. Those cell cycle molecules that are important for regulating terminal differentiation of cardiac myocytes and whether they can be targeted to reinitiate cell division and myocardial repair will be discussed as will the molecules that control vascular smooth muscle cell (VSMC) and endothelial cell proliferation in disorders such as atherosclerosis and restenosis. The main approaches currently used to target the cell cycle machinery in cardiovascular disease have employed gene therapy techniques. We will overview the different methods and routes of gene delivery to the cardiovascular system and describe possible future drug therapies for these disorders. Although the majority of the published data comes from animal studies, there are several instances where potential therapies have moved into the clinical setting with promising results.

Reprogramming the cell cycle machinery to treat cardiovascular disease

Coronary artery disease is one of the most common heart pathologies. Restriction of blood flow to the heart by atherosclerotic lesions, leading to angina pectoris and myocardial infarction, damages the heart, resulting in impaired cardiac function. Damaged myocardium is replaced by scar tissue since surviving cardiomyocytes are unable to proliferate to replace lost heart tissue. Although narrowing of the coronary arteries can be treated successfully using coronary revascularisation procedures, re-occlusion of the treated vessels remains a significant clinical problem. Cell cycle control mechanisms are key in both the impaired cardiac repair by surviving cardiomyocytes and re-narrowing of treated vessels by maladaptive proliferation of vascular smooth muscle cells. Strategies targeting the cell cycle machinery in the heart and vasculature offer promise both for the improvement of cardiac repair following MI and the prevention of restenosis and bypass graft failure following revascularisation procedures.

Multivariable cluster analysis for high-speed industrial machinery

The overall operation and internal complexity of a particular production machinery can be depicted in terms of clusters of multidimensional points which describe the process states, the value in each point dimension representing a measured variable from the machinery. The paper describes a new cluster analysis technique for use with manufacturing processes, to illustrate how machine behaviour can be categorised and how regions of good and poor machine behaviour can be identified. The cluster algorithm presented is the novel mean-tracking algorithm, capable of locating N-dimensional clusters in a large data space in which a considerable amount of noise is present. Implementation of the algorithm on a real-world high-speed machinery application is described, with clusters being formed from machinery data to indicate machinery error regions and error-free regions. This analysis is seen to provide a promising step ahead in the field of multivariable control of manufacturing systems.

Trypanosome Prereplication Machinery Contains a Single Functional Orc1/Cdc6 Protein, Which Is Typical of Archaea

In unicellular eukaryotes, such as Saccharomyces cerevisiae, and in multicellular organisms, the replication origin is recognized by the heterohexamer origin recognition complex (ORC) containing six proteins, Orc1 to Orc6, while in members of the domain Archaea, the replication origin is recognized by just one protein, Orc1/Cdc6; the sequence of Orc1/Cdc6 is highly related to those of Orc1 and Cdc6. Similar to Archaea, trypanosomatid genomes contain only one gene encoding a protein named Orc1. Since trypanosome Orc1 is also homologous to Cdc6, in this study we named the Orc1 protein from trypanosomes Orc1/Cdc6. Here we show that the recombinant Orc1/Cdc6 from Trypanosoma cruzi (TcOrc1/Cdc6) and from Trypanosoma brucei (TbOrc1/Cdc6) present ATPase activity, typical of prereplication machinery components. Also, TcOrc1/Cdc6 and TbOrc1/Cdc6 replaced yeast Cdc6 but not Orc1 in a phenotypic complementation assay. The induction of Orc1/Cdc6 silencing by RNA interference in T. brucei resulted in enucleated cells, strongly suggesting the involvement of Orc1/Cdc6 in DNA replication. Orc1/Cdc6 is expressed during the entire cell cycle in the nuclei of trypanosomes, remaining associated with chromatin in all stages of the cell cycle. These results allowed us to conclude that Orc1/Cdc6 is indeed a member of the trypanosome prereplication machinery and point out that trypanosomes carry a prereplication machinery that is less complex than other eukaryotes and closer to archaea.

State-of-the-art for extrusion and injection moulding FPC: natural Fibre Plastics Composites in Brazil

Environmental and economical concerns are stimulating research of new materials for construction, furniture, packaging and automotive industries. Particularly attractive are the new materials. This paper presents results about the properties of composites of polyhydroxybutyrate (PHB), polypropylene (PP) and High Density Polyethylene (HDPE) reinforced with natural fibres. The composites were prepared through the extrusion with natural fibres, in an ratio of 30% by weight. The quality of samples was tested utilising injection moulded samples, according to ASTM standards. Results showed that natural fibres can act as an excellent reinforcing material, resulting in low cost composites and improving economical and ecological interests.

eIF5A binds to translational machinery components and affects translation in yeast

The putative translation factor eIF5A is essential for cell viability and is highly conserved from archebacteria to mammals. Although this protein was originally identified as a translation initiation factor, subsequent experiments did not support a role for eIF5A in general translation. In this work, we demonstrate that eIF-5A interacts with structural components of the 80S ribosome, as well as with the translation elongation factor 2 (eEF2). Moreover, eIF5A is further shown to cofractionate with monosomes in a translation-dependent manner. Finally, eIF5A mutants show altered polysome profiles and are sensitive to translation inhibitors. Our results re-establish a function for eIF5A in translation and suggest a role for this factor in translation elongation instead of translation initiation. (c) 2006 Elsevier B.V. All rights reserved.

Different responses in bulk density and saturated hydraulic conductivity to soil deformation by logging machinery on a Ferralsol under native forest

Ferralsols have high structural stability, although structural degradation has been observed to result from forest to tillage or pasture conversion. An experimental series of forest skidder passes in an east Amazonian natural forest was performed for testing the effects of mechanical stress during selective logging operations on a clay-rich Ferralsol under both dry and wet soil conditions. Distinct ruts formed up to 25 cm depth only under wet conditions. After nine passes the initially very low surface bulk density of between 0.69 and 0.80 g cm(-3) increased to 1.05 g cm(-3) in the wet soil and 0.92 g cm(-3) in the dry soil. Saturated hydraulic conductivities, initially > 250 mm h(-1), declined to a minimum of around 10 mm h(-1) in the wet soil after the first pass, and in the dry soil more gradually after nine passes. The contrasting response of bulk density and saturated hydraulic conductivity is explained by exposure of subsoil material at the base of the ruts where macrostructure rapidly deteriorated under wet conditions. We attribute the resultant moderately high hydraulic conductivities to the formation of stable microaggregates with fine sand to coarse silt textures. We conclude that the topsoil macrostructure of Ferralsols is subject to similar deterioration to that of Luvisols in temperate zones. The stable microstructure prevents marked compaction and decrease in hydraulic conductivity under wetter and more plastic soil conditions. However, typical tropical storms may regularly exceed the infiltration capacity of the deformed soils. In the deeper ruts water may concentrate and cause surface run-off, even in gently sloping areas. To avoid soil erosion, logging operations in sloping areas should therefore be restricted to dry soil conditions when rut formation is minimal.

Alternatives Panels from Waste Plastics Reinforced With Fibers of Pupunha Palm for Use in Civil Construction and Furniture Industry

In order to cooperate in minimizing the problems of the current and growing volume of waste, this work aims at the production of panels made from industrial waste -thermoplastic (Polypropylene - PP; Polyethylene - PE and Acrylonitrile Butadiene Styrene - ABS) reinforced with agro-industrial waste - pupunha palm waste (shells and sheaths). The properties of the panels were evaluated: density, thickness swelling, water absorption and moisture content. It was used the ASTM D1037; EN 317; and ANSI A208.1 standards regarding particle boards. The best results in physical tests were treatments 1 (100% waste plastic), 6 (60% plastic waste and 40% waste of pupunha) and 7 (70% waste plastic and 30% waste of pupunha). The best results in the mechanical tests were treatments 3 (30% de residuos plasticos e 70% de residuos da pupunha), 4 (40% de residuos plasticos c 60% de residuos da pupunha) and 5 (50% de residuos plasticos e 50% de residuos da pupunha). For mechanical tests it was concluded that the results of modulus of rupture and of modulus of elasticity the best treatments were those with more fibers. In the tensile tests perpendicular to the surface, it is clear that using more waste plastics leads to the best results. It was concluded that the waste can be used as raw material for the production of alternative materials mainly in civil construction and furniture industries, and it can be employed in urban or rural environment, given the concept of eco-efficient products.

Active control in rotating machinery using magnetic actuators with linear matrix inequalities (LMI) approach

The recent years have seen the appearance of innovative system for acoustic and vibration attenuation, most of them integrating new actuator technologies. In this sense, the study of algorithms for active vibrations control in rotating machinery became an area of enormous interest, mainly due to countless demands of an optimal performance of mechanical systems in aircraft, aerospace and automotive structures. In this way, this paper presents an approach that is numerically verified for active vibration control in a rotor using Active Magnetic Bearings (AMB). The control design in a discrete state-space formulation is carried out through feedback technique and Linear Matrix Inequalities (LMI) approach. LMI is useful for system with uncertainties. The AMB uses electromagnetic forces to support a rotor without mechanical contact. By monitoring the position of the shaft and changing the dynamics of the system accordingly, the AMB keeps the rotor in a desired position. This unique feature has broadened for the applications of AMB and now they can be considered not only as a main support bearing in a machine but also as dampers for vibration control and force actuators. © 2009 Society for Experimental Mechanics Inc.