Differential induction of 12-O-tetradecanoylphorbol 13-acetate sequence gene expression in murine melanocytes and melanoma cells

We previously showed that growth of the nontumorigenic, immortal murine melanocyte line Mel-ab correlates with the depletion of protein kinase C (PKC), whereas quiescence is associated with elevated levels of this enzyme (Brooks G, et al., Cancer Res 51: 3281–3288, 1991). Here we report responses that occur in these cells downstream of PKC activation or downregulation. We examined induction of 12-O-tetradecanoylphorbol-13-acetate (TPA)-inducible sequence (TIS) gene expression in Mel-ab melanocytes and in their transformed counterparts, B16 melanoma cells. Exposure of quiescent Mel-ab cells to the PKC-activating phorbol esters TPA or sapintoxin A at 81 nM for 2 h increased levels of mRNA for six of seven TIS genes examined (twofold to 80-fold increase in steady-state RNA levels for TIS 1, 7, 8, 11, 21, and 28 (c-fos); TIS 10 expression was not affected). No induction of 115 gene expression was observed either in growing Mel-ab cells maintained in 324 nM phorbol 12,13-dibutyrate or in B16 cells previously unexposed to phorbol esters, in which normal PKC levels were endogenously depressed. The cAMP-elevating agents choleratoxin (10 nM) and dibutyryl cyclic AMP (2.5 mM) increased levels of TIS mRNA (with the exception of TIS 10) in both proliferating Mel-ab and B16 cells, suggesting that downregulation of the PKC pathway is specific and not a consequence of a general inhibition of all signalling pathways.

Speed sensorless vector control of induction machines

A novel rotor velocity estimation scheme applicable to vector controlled induction motors has been described. The proposed method will evaluate rotor velocity, ωr, on-line, does not require any extra transducers or injection of any signals, nor does it employ complicated algorithms such as MRAS or Kalman filters. Furthermore, the new scheme will operate at all velocities including zero with very little error. The procedure employs motor model equations, however all differential and integral terms have been eliminated giving a very fast, low-cost, effective and practical alternative to the current available methods. Simulation results verify the operation of the scheme under ideal and PWM conditions.

Differential recognition of Staphylococcus aureus quorum-sensing signals depends on both extracellular loops 1 and 2 of the transmembrane sensor AgrC.

Virulence in Staphylococcus aureus is regulated via agr-dependent quorum sensing in which an autoinducing peptide (AIP) activates AgrC, a histidine protein kinase. AIPs are usually thiolactones containing seven to nine amino acid residues in which the thiol of the central cysteine is linked to the alpha-carboxyl of the C-terminal amino acid residue. The staphylococcal agr locus has diverged such that the AIPs of the four different S. aureus agr groups self-activate but cross-inhibit. Consequently, although the agr system is conserved among the staphylococci, it has undergone significant evolutionary divergence whereby to retain functionality, any changes in the AIP-encoding gene (agrD) that modifies AIP structure must be accompanied by corresponding changes in the AgrC receptor. Since AIP-1 and AIP-4 only differ by a single amino acid, we compared the transmembrane topology of AgrC1 and AgrC4 to identify amino acid residues involved in AIP recognition. As only two of the three predicted extracellular loops exhibited amino acid differences, site-specific mutagenesis was used to exchange the key AgrC1 and AgrC4 amino acid residues in each loop either singly or in combination. A novel lux-based agrP3 reporter gene fusion was constructed to evaluate the response of the mutated AgrC receptors. The data obtained revealed that while differential recognition of AIP-1 and AIP-4 depends primarily on three amino acid residues in loop 2, loop 1 is essential for receptor activation by the cognate AIP. Furthermore, a single mutation in the AgrC1 loop 2 resulted in conversion of (Ala5)AIP-1 from a potent antagonist to an activator, essentially resulting in the forced evolution of a new AIP group. Taken together, our data indicate that loop 2 constitutes the predicted hydrophobic pocket that binds the AIP thiolactone ring while the exocyclic amino acid tail interacts with loop 1 to facilitate receptor activation.

Making DNA without iron: induction of a manganese-dependent ribonucleotide reductase in response to iron starvation

Ribonucleotide reductases supply cells with their deoxyribonucleotides. Three enzyme types are known, classes I, II and III. Class II enzymes are anaerobic whereas class I enzymes are aerobic, and so class I and II enzymes are often produced by the same organism under opposing oxygen regimes. Escherichia coli contains two types of class I enzyme (Ia and Ib) with the Fe-dependent Ia enzyme (NrdAB) performing the major role aerobically, leaving the purpose of the Ib enzyme (NrdEF) unclear. Several papers have recently focused on the class Ib enzymes showing that they are Mn (rather than Fe) dependent and suggesting that the E. coli NrdEF may function under redox-stress conditions. A paper published in this issue of Molecular Microbiology from James Imlay's group confirms that this unexplained NrdEF Ib enzyme is Mn-dependent, but shows that it does not substitute for NrdAB during redox stress. Instead, a role during iron restriction is demonstrated. Thus, the purpose of NrdEF (and possibly other class Ib enzymes) is to enhance growth under aerobic, low-iron conditions, and to functionally replace the Fe-dependent NrdAB when iron is unavailable. This finding reveals a new mechanism by which bacteria adjust to life under iron deprivation.

Induction of the ferritin gene (ftnA) of Escherichia coli by Fe2+–Fur is mediated by reversal of H-NS silencing and is RyhB independent

FtnA is the major iron-storage protein of Escherichia coli accounting for < or = 50% of total cellular iron. The FtnA gene (ftnA) is induced by iron in an Fe(2+)-Fur-dependent fashion. This effect is reportedly mediated by RyhB, the Fe(2+)-Fur-repressed, small, regulatory RNA. However, results presented here show that ftnA iron induction is independent of RyhB and instead involves direct interaction of Fe(2+)-Fur with an 'extended' Fur binding site (containing five tandem Fur boxes) located upstream (-83) of the ftnA promoter. In addition, H-NS acts as a direct repressor of ftnA transcription by binding at multiple sites (I-VI) within, and upstream of, the ftnA promoter. Fur directly competes with H-NS binding at upstream sites (II-IV) and consequently displaces H-NS from the ftnA promoter (sites V-VI) which in turn leads to derepression of ftnA transcription. It is proposed that H-NS binding within the ftnA promoter is facilitated by H-NS occupation of the upstream sites through H-NS oligomerization-induced DNA looping. Consequently, Fur displacement of H-NS from the upstream sites prevents cooperative H-NS binding at the downstream sites within the promoter, thus allowing access to RNA polymerase. This direct activation of ftnA transcription by Fe(2+)-Fur through H-NS antisilencing represents a new mechanism for iron-induced gene expression.

The role of myeloid receptors on murine plasmacytoid dendritic cells in induction of type I interferon

This study tested the hypothesis that a set of predominantly myeloid restricted receptors (F4/80, CD36, Dectin-1, CD200 receptor and mannan binding lectins) and the broadly expressed CD200 played a role in a key function of plasmacytoid DC (pDC), virally induced type I interferon (IFN) production. The Dectin-1 ligands zymosan, glucan phosphate and the anti-Dectin-1 monoclonal antibody (mAb) 2A11 had no effect on influenza virus induced IFNα/β production by murine splenic pDC. However, mannan, a broad blocking reagent against mannose specific receptors, inhibited IFNα/β production by pDC in response to inactivated influenza virus. Moreover, viral glycoproteins (influenza virus haemagglutinin and HIV-1 gp120) stimulated IFNα/β production by splenocytes in a mannan-inhibitable manner, implicating the function of a lectin in glycoprotein induced IFN production. Lastly, the effect of CD200 on IFN induction was investigated. CD200 knock-out macrophages produced more IFNα than wild-type macrophages in response to polyI:C, a MyD88-independent stimulus, consistent with CD200's known inhibitory effect on myeloid cells. In contrast, blocking CD200 with an anti-CD200 mAb resulted in reduced IFNα production by pDC-containing splenocytes in response to CpG and influenza virus (MyD88-dependent stimuli). This suggests there could be a differential effect of CD200 on MyD88 dependent and independent IFN induction pathways in pDC and macrophages. This study supports the hypothesis that a mannan-inhibitable lectin and CD200 are involved in virally induced type I IFN induction.

Dynamic simulation for closed loops in heating systems with decision-making support

Nowadays utilising the proper HVAC system is essential both in extreme weather conditions and dense buildings design. Hydraulic loops are the most common parts in all air conditioning systems. This article aims to investigate the performance of different hydraulic loop arrangements in variable flow systems. Technical, economic and environmental assessments have been considered in this process. A dynamic system simulation is generated to evaluate the system performance and an economic evaluation is conducted by whole life cost assessment. Moreover, environmental impacts have been studied by considering the whole life energy consumption, CO2 emission, the embodied energy and embodied CO2 of the system components. Finally, decision-making in choosing the most suitable hydraulic system among five well-known alternatives has been proposed.

Steering without Circe : attending to reinforcing loops in social systems

Relating system dynamics to the broad systems movement, the key notion is that reinforcing loops deserve no less attention than balancing loops. Three specific propositions follow. First, since reinforcing loops arise in surprising places, investigations of complex systems must consider their possible existence and potential impact. Second, because the strength of reinforcing loops can be misinferred - we include an example from the field of servomechanisms - computer simulation can be essential. Be it project management, corporate growth or inventory oscillation, simulation helps to assess consequences of reinforcing loops and options for interventions. Third, in social systems the consequences of reinforcing loops are not inevitable. Examples concerning globalization illustrate how difficult it might be to challenge such assumptions. However, system dynamics and ideas from contemporary social theory help to show that even the most complex social systems are, in principle, subject to human influence. In conclusion, by employing these ideas, by attending to reinforcing as well as balancing loops, system dynamics work can improve the understanding of social systems and illuminate our choices when attempting to steer them.

PMCRI: a parallel modular classification rule induction framework

In a world where massive amounts of data are recorded on a large scale we need data mining technologies to gain knowledge from the data in a reasonable time. The Top Down Induction of Decision Trees (TDIDT) algorithm is a very widely used technology to predict the classification of newly recorded data. However alternative technologies have been derived that often produce better rules but do not scale well on large datasets. Such an alternative to TDIDT is the PrismTCS algorithm. PrismTCS performs particularly well on noisy data but does not scale well on large datasets. In this paper we introduce Prism and investigate its scaling behaviour. We describe how we improved the scalability of the serial version of Prism and investigate its limitations. We then describe our work to overcome these limitations by developing a framework to parallelise algorithms of the Prism family and similar algorithms. We also present the scale up results of a first prototype implementation.

Parallel induction of modular classification rules

The Distributed Rule Induction (DRI) project at the University of Portsmouth is concerned with distributed data mining algorithms for automatically generating rules of all kinds. In this paper we present a system architecture and its implementation for inducing modular classification rules in parallel in a local area network using a distributed blackboard system. We present initial results of a prototype implementation based on the Prism algorithm.

Parallel rule induction with information theoretic pre-pruning

In a world where data is captured on a large scale the major challenge for data mining algorithms is to be able to scale up to large datasets. There are two main approaches to inducing classification rules, one is the divide and conquer approach, also known as the top down induction of decision trees; the other approach is called the separate and conquer approach. A considerable amount of work has been done on scaling up the divide and conquer approach. However, very little work has been conducted on scaling up the separate and conquer approach.In this work we describe a parallel framework that allows the parallelisation of a certain family of separate and conquer algorithms, the Prism family. Parallelisation helps the Prism family of algorithms to harvest additional computer resources in a network of computers in order to make the induction of classification rules scale better on large datasets. Our framework also incorporates a pre-pruning facility for parallel Prism algorithms.

P-Prism: a computationally efficient approach to scaling up classification rule induction

Top Down Induction of Decision Trees (TDIDT) is the most commonly used method of constructing a model from a dataset in the form of classification rules to classify previously unseen data. Alternative algorithms have been developed such as the Prism algorithm. Prism constructs modular rules which produce qualitatively better rules than rules induced by TDIDT. However, along with the increasing size of databases, many existing rule learning algorithms have proved to be computational expensive on large datasets. To tackle the problem of scalability, parallel classification rule induction algorithms have been introduced. As TDIDT is the most popular classifier, even though there are strongly competitive alternative algorithms, most parallel approaches to inducing classification rules are based on TDIDT. In this paper we describe work on a distributed classifier that induces classification rules in a parallel manner based on Prism.