Characterisation of the putative effector interaction site of the regulatory HbpR protein from Pseudomonas azelaica by site-directed mutagenesis.

Bacterial transcription activators of the XylR/DmpR subfamily exert their expression control via σ(54)-dependent RNA polymerase upon stimulation by a chemical effector, typically an aromatic compound. Where the chemical effector interacts with the transcription regulator protein to achieve activation is still largely unknown. Here we focus on the HbpR protein from Pseudomonas azelaica, which is a member of the XylR/DmpR subfamily and responds to biaromatic effectors such as 2-hydroxybiphenyl. We use protein structure modeling to predict folding of the effector recognition domain of HbpR and molecular docking to identify the region where 2-hydroxybiphenyl may interact with HbpR. A large number of site-directed HbpR mutants of residues in- and outside the predicted interaction area was created and their potential to induce reporter gene expression in Escherichia coli from the cognate P(C) promoter upon activation with 2-hydroxybiphenyl was studied. Mutant proteins were purified to study their conformation. Critical residues for effector stimulation indeed grouped near the predicted area, some of which are conserved among XylR/DmpR subfamily members in spite of displaying different effector specificities. This suggests that they are important for the process of effector activation, but not necessarily for effector specificity recognition.

Tumor targeting with newly designed biparatopic antibodies directed against two different epitopes of the carcinoembryonic antigen (CEA).

In an attempt to improve tumor targeting and tumor retention time of monoclonal antibodies (MAbs), we prepared biparatopic antibodies (BpAbs) having the capability of binding 2 different non-overlapping epitopes on the same target antigen molecule, namely, the carcinoembryonic antigen (CEA). Six BpAbs were constructed by coupling 2 different Fab' fragments from 4 different specific anti-CEA MAbs recognizing 4 CEA epitopes (Gold 1-4). Demonstration of the double paratopic binding of these antibodies for CEA was confirmed in vitro by inhibition radioimmunoassay and cross-inhibition analysis by surface plasmon resonance (SPR; BIACORE) technology. Using the latter technique, the affinity constants for CEA immobilized onto the sensor chip were found to range from 0.37 to 1.54 x 10(9) M(-1) for the 4 parental F(ab')2 fragments and from 1.88 to 10.14 x 10(9) M(-1) for the BpAbs, demonstrating the advantage of biparatopic binding over conventional F(ab')2 binding. The Ka improvement was particularly high for BpAb F6/35A7 and BpAb F6/B17 with a 9.5- and 8.1-fold increase, respectively, as compared with the parental F(ab')2. In vivo, the 6 BpAbs were compared with their 2 respective parental F(ab')2 by injection of 131I-BpAb/125I-F(ab')2 parental fragments into nude mice xenografted with the human colon carcinoma T380. Dissection 72 hr post-injection demonstrated that BpAb B17/CE25 and BpAb F6/B17 gave higher tumor uptake than that of their parental F(ab')2. This finding is particularly interesting for BpAb F6/B17, which compared favorably with the F6 F(ab')2, one of the best parental F(ab')2 fragments used in our study.

Chaotic systems with a null conditional Lyapunov exponent under nonlinear driving.

Control of a chaotic system by homogeneous nonlinear driving, when a conditional Lyapunov exponent is zero, may give rise to special and interesting synchronizationlike behaviors in which the response evolves in perfect correlation with the drive. Among them, there are the amplification of the drive attractor and the shift of it to a different region of phase space. In this paper, these synchronizationlike behaviors are discussed, and demonstrated by computer simulation of the Lorentz model [E. N. Lorenz, J. Atmos. Sci. 20 130 (1963)] and the double scroll [T. Matsumoto, L. O. Chua, and M. Komuro, IEEE Trans. CAS CAS-32, 798 (1985)].

Alternative tests for correct specification of conditional predictive densities

We propose new methods for evaluating predictive densities that focus on the models' actual predictive ability in finite samples. The tests offer a simple way of evaluatingthe correct specification of predictive densities, either parametric or non-parametric.The results indicate that our tests are well sized and have good power in detecting mis-specification in predictive densities. An empirical application to the Survey ofProfessional Forecasters and a baseline Dynamic Stochastic General Equilibrium modelshows the usefulness of our methodology.

Scheduling an European basketball competition. An application of graph theory

The object of this project is to schedule a ctitious European basketball competition with many teams situated a long distances. The schedule must be fair, feasible and economical, which means that the total distance trav- eled by every team must be the minimal possible. First, we de ne the sport competition terminology and study di erent competition systems, focusing on the NBA and the Euroleague systems. Then we de ne concepts of graph theory and spherical distance that will be needed. Next we propose a com- petition system, explaining where will be allocated the teams and how will be the scheduling. Then there is a description of the programs that have been implemented, and, nally, the complete schedule is displayed, and some possible improvements are mentioned.

Conditional equilibrium constants in multicomponent heterogeneous adsorption: The conditional affinity spectrum

The concept of conditional stability constant is extended to the competitive binding of small molecules to heterogeneous surfaces or macromolecules via the introduction of the conditional affinity spectrum (CAS). The CAS describes the distribution of effective binding energies experienced by one complexing agent at a fixed concentration of the rest. We show that, when the multicomponent system can be described in terms of an underlying affinity spectrum [integral equation (IE) approach], the system can always be characterized by means of a CAS. The thermodynamic properties of the CAS and its dependence on the concentration of the rest of components are discussed. In the context of metal/proton competition, analytical expressions for the mean (conditional average affinity) and the variance (conditional heterogeneity) of the CAS as functions of pH are reported and their physical interpretation discussed. Furthermore, we show that the dependence of the CAS variance on pH allows for the analytical determination of the correlation coefficient between the binding energies of the metal and the proton. Nonideal competitive adsorption isotherm and Frumkin isotherms are used to illustrate the results of this work. Finally, the possibility of using CAS when the IE approach does not apply (for instance, when multidentate binding is present) is explored. © 2006 American Institute of Physics.

An open source browser-based software tool for graph drawing and visualisation

In this research work we searched for open source libraries which supports graph drawing and visualisation and can run in a browser. Subsequent these libraries were evaluated to find out which one is the best for this task. The result was the d3.js is that library which has the greatest functionality, flexibility and customisability. Afterwards we developed an open source software tool where d3.js was included and which was written in JavaScript so that it can run browser-based.

WebGraphEd, an open source graph drawing editor for the web

WebGraphEd is an open source software for graph visualization and manipulation. It is especially designed to work for the web platform through a web browser. The web application has been written in JavaScript and compacted later, which makes it a very lightweight software. There is no need of additional software, and the only requirement is to have an HTML5 compliant browser. WebGraphEd works with scalable vector graphics (SVG), which it makes possible to create lossless graph drawings.

A note on the periodic orbits and topological entropy of graph maps

This paper deals with the relationship between the periodic orbits of continuous maps on graphs and the topological entropy of the map. We show that the topological entropy of a graph map can be approximated by the entropy of its periodic orbits.

Laajakaistaverkon topologian visualisointi

Viimeisten vuosien aikana laajakaistaoperaattoreiden laajakaistaverkot ovat nopeiden ja kiinteähintaisten laajakaistaliittymien johdosta kasvaneet suuriksi kokonaisuuksiksi. Kokonaisuuksia hallitaan erilaisilla verkonhallintatyökaluilla. Verkonhallintatyökalut sisältävät suuren määrän eri tasoista tietoa laitteista ja laitteiden välisistä suhteista. Kokonaisuuksien hahmottaminen ilman tiedoista rakennettua kuvaa on vaikeaa ja hidasta. Laajakaistaverkon topologian visualisoinnissa muodostetaan kuva laitteista ja niiden välisistä suhteista. Visualisoitua kuvaa voidaan käyttää osana verkonhallintatyökalua, jolloin käyttäjälle muodostuu nopeasti näkymä verkon laitteista ja rakenteesta eli topologiasta. Visualisoinnissa kuvan piirto-ongelma täytyy muuttaa graafin piirto-ongelmaksi. Graafin piirto-ongelmassa verkon rakennetta käsitellään graafina, joka mahdollistaa kuvan muodostamisen automaattisia piirtomenetelmiä hyväksikäyttäen. Halutunlainen ulkoasu kuvalle muodostetaan automaattisilla piirtomenetelmillä, joilla laitteiden ja laitteiden välisten suhteiden esitystapoja voidaan muuttaa. Esitystavoilla voidaan muuttaa esimerkiksi laitteiden muotoa, väriä ja kokoa. Esitystapojen lisäksi piirtomenetelmien tärkein tehtävä on laskea laitteiden sijaintien koordinaattien arvot, jotka loppujen lopuksi määräävät koko kuvan rakenteen. Koordinaattien arvot lasketaan piirtoalgoritmeilla, joista voimiin perustuvat algoritmit sopivat parhaiten laajakaistaverkkojen laitteiden sijaintien laskemiseen. Tämän diplomityön käytännön työssä toteutettiin laajakaistaverkon topologian visualisointityökalu.

Social network extraction and analysis based on multimodal dyadic interaction

Social interactions are a very important component in people"s lives. Social network analysis has become a common technique used to model and quantify the properties of social interactions. In this paper, we propose an integrated framework to explore the characteristics of a social network extracted from multimodal dyadic interactions. For our study, we used a set of videos belonging to New York Times" Blogging Heads opinion blog. The Social Network is represented as an oriented graph, whose directed links are determined by the Influence Model. The links" weights are a measure of the"influence" a person has over the other. The states of the Influence Model encode automatically extracted audio/visual features from our videos using state-of-the art algorithms. Our results are reported in terms of accuracy of audio/visual data fusion for speaker segmentation and centrality measures used to characterize the extracted social network.