RNAi, DRD1, and histone methylation actively target developmentally important Non-CG DNA methylation in Arabidopsis

Cytosine DNA methylation protects eukaryotic genomes by silencing transposons and harmful DNAs, but also regulates gene expression during normal development. Loss of CG methylation in the Arabidopsis thaliana met1 and ddm1 mutants causes varied and stochastic developmental defects that are often inherited independently of the original met1 or ddm1 mutation. Loss of non-CG methylation in plants with combined mutations in the DRM and CMT3 genes also causes a suite of developmental defects. We show here that the pleiotropic developmental defects of drm1 drm2 cmt3 triple mutant plants are fully recessive, and unlike phenotypes caused by met1 and ddm1, are not inherited independently of the drm and cmt3 mutations. Developmental phenotypes are also reversed when drm1 drm2 cmt3 plants are transformed with DRM2 or CMT3, implying that non-CG DNA methylation is efficiently re-established by sequence-specific signals. We provide evidence that these signals include RNA silencing though the 24-nucleotide short interfering RNA (siRNA) pathway as well as histone H3K9 methylation, both of which converge on the putative chromatin-remodeling protein DRD1. These signals act in at least three partially intersecting pathways that control the locus-specific patterning of non-CG methylation by the DRM2 and CMT3 methyltransferases. Our results suggest that non-CG DNA methylation that is inherited via a network of persistent targeting signals has been co-opted to regulate developmentally important genes. © 2006 Chan et al.

How to avoid the problems of target-setting

Purpose: Advocates and critics of target-setting in the workplace seem unable to reach beyond their own well-entrenched battle lines. While the advocates of goal-directed behaviour point to what they see as demonstrable advantages, the critics of target-setting highlight equally demonstrable disadvantages. Indeed, the academic literature on this topic is currently mired in controversy, with neither side seemingly capable of envisaging a better way forward. This paper seeks to break the current deadlock and move thinking forward in this important aspect of performance measurement and management by outlining a new, more fruitful approach, based on both theory and practical experience. Design/methodology/approach: The topic was approached in three phases: assembling and reading key academic and other literature on the subject of target-setting and goal-directed behaviour, with a view to understanding, in depth, the arguments advanced by the advocates and critics of target-setting; comparing these published arguments with one's own experiential findings, in order to bring the essence of disagreement into much sharper focus; and then bringing to bear the academic and practical experience to identify the essential elements of a new, more fruitful approach offering all the benefits of goal-directed behaviour with none of the typical disadvantages of target-setting. Findings: The research led to three key findings: the advocates of goal-directed behaviour and critics of target-setting each make valid points, as seen from their own current perspectives; the likelihood of these two communities, left to themselves, ever reaching a new synthesis, seems vanishingly small (with leading thinkers in the goal-directed behaviour community already acknowledging this); and, between the three authors, it was discovered that their unusual combination of academic study and practical experience enabled them to see things differently. Hence, they would like to share their new thinking more widely. Research limitations/implications: The authors fully accept that their paper is informed by extensive practical experience and, as yet, there have been no opportunities to test their findings, conclusions and recommendations through rigorous academic research. However, they hope that the paper will move thinking forward in this arena, thereby informing future academic research. Practical implications: The authors hope that the practical implications of the paper will be significant, as it outlines a novel way for organisations to capture the benefits of goal-directed behaviour with none of the disadvantages typically associated with target-setting. Social implications: Given that increased efficiency and effectiveness in the management of organisations would be good for society, the authors think the paper has interesting social implications. Originality/value: Leading thinkers in the field of goal-directed behaviour, such as Locke and Latham, and leading critics of target-setting, such as Ordóñez et al. continue to argue with one another - much like, at the turn of the nineteenth century, proponents of the "wave theory of light" and proponents of the "particle theory of light" were similarly at loggerheads. Just as this furious scientific debate was ultimately resolved by Taylor's experiment, showing that light could behave both as a particle and wave at the same time, the authors believe that the paper demonstrates that goal-directed behaviour and target-setting can successfully co-exist. © Emerald Group Publishing Limited.

Calcium-permeable acid-sensing ion channel is a molecular target of the neurotoxic metal ion lead

Acid-sensing ion channels (ASICs) are emerging as fundamental players in the regulation of neural plasticity and in pathological conditions. Here we showed that lead (Pb2+), a well known neurotoxic metal ion, reversibly and concentration-dependently inhib

Cloning and expression of zebra fish ferroportin and investigating its influence on anemia

Iron is required for many microbes and pathogens for their survival and proliferation including Leishmania which cause leishmaniasis. Leishmaniasis is an increasingly serious infectious disease with a wide spectrum of clinical manifestations. These range from localized cutaneous leishmaniasis (CL) lesions to a lethal visceral form. Certain strains such as BALB/c mice fail to control L. major infection and develop progressive lesions and systemic disease. These mice are thought to be a model of non-healing forms of the human disease such as kala-azar or diffuse cutaneous leishmaniasis. Progression of disease in BALB/c mice has been associated with the anemia, in last days of their survival, the progressive anemia is considered to be one of the reasons of their death. Ferroportin (Fpn), a key regulator of iron homeostasis is a conserved membrane protein that exports iron across the duodenal enterocytes as well as macrophages and hepatocytes into the blood circulation. Fpn has also critical influence on survival and proliferation of many microorganisms whose growth is dependent upon iron, thus preparation of Fpn is needed to study the role of iron in immune responses and pathogenesis of micoorganisms. To prepare and characterize a recombinant ferroportin, total RNA was extracted from Indian zebrafish duodenum, and used to synthesize cDNA by RT-PCR. PCR product was first cloned in Topo TA vector and then subcloned into the GFP expression vector pEGFP–N1. The final resulted plasmid (pEGFP-ZFpn) was used for expression of FPN-EGFP protein in Hek 293T cells. The expression was confirmed by fluorescence microscopy and flow cytometery. Recombinant Fpn was further characterized by submission of its predicted amino acid sequences to the TMHMM V2.0 prediction server (hidden Markov model), NetOGlyc 3.1 server and NetNGlyc 3.1 server. Data emphasised that obtained Fpn from indian zebrafish contained eight transmembrane domains with N- and C-termini inside the cytoplasm and harboured 78 mucin-type glycosylated amino acid. The results indicate that the prepared and characterized recombinant Fpn protein has no membrane topology difference compared to other Fpn described by other researcher. Our next aim was to deliver recombinant plasmid (pEGFP-ZFpn) to entrocyte cells. However, naked therapeutic genes are rapidly degraded by nucleases, showing poor cellular uptake, nonspecificity to the target cells, and low transfection efficiency. The development of safe and efficient gene carriers is one of the prerequisites for the success of gene therapy. Chitosan and alginate 139 polymers were used for oral gene carrier because of their biodegradability, biocompatibility and their mucoadhesive and permeability-enhancing properties in the gut. Nanoparticles comprising Alginate/Chitosan polymers were prepared by pregel preparation method. The resulting nanoparticles had a loading efficiency of 95% and average size of 188 nm as confirmed by PCS method and SEM images had showed spherical particles. BALB/c mice were divided to three groups. The first and second group were fed with chitosan/alginate nanoparticles containing the pEGFP-ZFpn and pEGFP plasmid, respectively (30 μgr/mice) and the third group (control) didn’t get any nanoparticles. The result showed BALB/c mice infected by L.major, resulted in higher hematocryte and iron level in pEGFP-ZFpn fed mice than that in other groups. Consentration of cytokines determined by ELISA showed lower levels of IL-4 and IL-10 and higher levels of IFN-γ/IL-4 and IFN-γ/IL-10 ratios in pEGFP-ZFpn fed mice than that in other groups. Morover more limited increase of footpad thickness and significant reduction of viable parasites in lymph node was seen in pEGFP-ZFpn fed mice. The results showed the first group exhibited a highr hematocryte and iron compared to the other groups. These data strongly suggests the in vivo administration of chitosan/alginate nanoparticles containing pEGFP-ZFpn suppress Th2 response and may be used to control the leishmaniasis .

Identification of triosephosphate isomerase (TIM) genes from Microcystis (Cyanobacteria : Chroococcales) and theoretical analyses of the potential of cyanobacterial TIM as a target for designing specific inhibitors

The genes encoding triosephosphate isomerase (TIM) in three species of Microcystis (M. aeruginosa, M. viridis and M. wesenbergii) were investigated. Reverse transcriptase-polymerase chain reaction indicated that they were transcribed in the cells. Analyses showed that their DNA and deduced amino acid sequences were highly conserved between all the three species, only a single nonsynonymous substitution was seen at position 31, from an Asp in M. aeruginosa and M. viridis to Glu in M. wesenbergii. Sequence alignment of these with 12 other known cyanobacterial TIM sequences showed that all the cyanobacterial TIMs had a very high level of amino acid identity (over 50% between each two). Comparison of the cyanobacterial TIMs with other reported TIMs (from diverse lineages of the three Domains) showed that they possessed common active-site residues and sequence motifs. All cyanobacterial TIMs have two common cysteine residues (Cys127 and Cys176), and the Cys176 is almost cyanobacteria-specific with only one exception in Streptomyces coelicolor. Both secondary structure alignment and comparative modelling of Synechocystis sp. TIM showed that Cys176 was located at the hinge region of the flexible loop-6 and might therefore be critical to the movement of TIM's loop-6, which is important to the function of the enzyme. Thus, the cyanobacterial TIM-specific Cys176 may be a potential site for the discovery of suitable drugs against cyanobacteria, and such drugs may have utility in controlling water blooms due to cyanobacteria.

Developing a technology intelligence strategy at kodak european research: Scan & target

OVERVIEW: Kodak European Research (KER) developed a strategy for technology intelligence based on a theoretical model developed by Kerr et al. (2006). KER scouts designed and implemented a four-step approach to identify relevant technologies and research centers across Europe, Africa and the Middle East. The approach provides clear guidance for integrating web searches, scouting trips, networking and interactions with intermediaries. KER's example illustrates how companies can organize themselves to look outside corporate boundaries in search of technologies relevant for their business. The approach may be useful to those in other companies who have been asked to start a technology intelligence activity. © 2010 Industrial Research Institute, Inc.

TMVA, a novel GPIb-binding protein, significantly prevents platelet microthrombi formation and prolongs discordant cardiac xenograft survival

In xenotransplantation, donor endothelium is the first target of immunological attack. Activation of the endothelial cell by preformed natural antibodies leads to platelet binding via the interaction of the glycoprotein (GP) Ib and von Willebrand factor (vWF). TMVA is a novel GPIb-binding protein purified from the venom of Trimeresurus mucrosquamatus. In this study, the inhibitory effect of TMVA on platelet aggregation in rats and the effect on discordant guinea pig-to-rat cardiac xenograft survival were investigated. Three doses (8, 20 or 40 mug/kg) of TMVA were infused intravenously to 30 rats respectively. Platelet aggregation rate was assayed 0.5, 12, and 24 h after TMVA administration. Wister rats underwent guinea pig cardiac cervical heterotopic transplantation using single dosing of TMVA (20 mug/kg, i.v., 0.5 h before reperfusion). Additionally, levels of TXB2 and 6-keto-PGF(1alpha) within rejected graft tissues were determined by radioimmunoassay. Treatment with TMVA at a dose of 20 or 40 mug/kg resulted in complete inhibition of platelet aggregation 0.5 h after TMVA administration. Rats receiving guinea pig cardiac xenografts after TMVA therapy had significantly prolonged xenograft survival. Histologic and immunopathologic analysis of cardiac xenografts in TMVA treatment group showed no intragraft platelet microthrombi formation and fibrin deposition. Additionally, the ratio of 6-keto-PGF(1alpha) to TXB2 in TMVA treatment group was significantly higher than those in control group. We conclude that the use of this novel GPIb-binding protein was very effective in preventing platelet microthrombi formation and fibrin deposition in a guinea pig-to-rat model and resulted in prolongation of xenograft survival. The increased ratio of PGI(2)/TXA(2) in TMVA treatment group may protect xenografts from the endothelial cell activation and contribute to the prolongation of xenograft survival.

A Monte Carlo expectation maximisation algorithm for multiple target tracking

In this paper, we present an expectation-maximisation (EM) algorithm for maximum likelihood estimation in multiple target models (MTT) with Gaussian linear state-space dynamics. We show that estimation of sufficient statistics for EM in a single Gaussian linear state-space model can be extended to the MTT case along with a Monte Carlo approximation for inference of unknown associations of targets. The stochastic approximation EM algorithm that we present here can be used along with any Monte Carlo method which has been developed for tracking in MTT models, such as Markov chain Monte Carlo and sequential Monte Carlo methods. We demonstrate the performance of the algorithm with a simulation. © 2012 ISIF (Intl Society of Information Fusi).

A preliminary study of target multiplicity for ADSRs

The Accelerator Driven Subcritical Reactor (ADSR) concept is based on the coupling of a particle accelerator to a subcritical reactor core by means of a neutron spallation target interface. This paper investigates the benefits of multiple spallation targets in ADSRs. The motivation behind this is, firstly, to improve the overall reliability of the accelerator-reactor system, and, secondly, to evaluate other potential advantages such as lower beam power requirements. The results show that a system containing two or three spallation targets, coupled to independent accelerators, offers better neutronic performance. This is demonstrated through the increased effective multiplication factor (keff) in the two- and three-target configurations and a more uniform neutron flux distribution. A multiple-target ADSR also proves effective in mitigating the impact of frequent beam interruptions, a pressing issue that needs to be addressed for the ADSR concept to advance. Assuming no simultaneous beam shutdowns, the two- and three-target configurations reduce the risk of fuel cladding failure due to thermal cyclic fatigue. © 2013 Elsevier B.V. All rights reserved.

User target intention recognition from cursor position using kalman filter

This paper discusses user target intention recognition algorithms for pointing - clicking tasks to reduce users' pointing time and difficulty. Predicting targets by comparing the bearing angles to targets proposed as one of the first algorithms [1] is compared with a Kalman Filter prediction algorithm. Accuracy and sensitivity of prediction are used as performance criteria. The outcomes of a standard point and click experiment are used for performance comparison, collected from both able-bodied and impaired users. © 2013 Springer-Verlag Berlin Heidelberg.