Generation and characterisation of acute myeloid leukaemia-reactive CD4 + T cells and their potential to eliminate human leukaemia blasts in NSG mice

Acute myeloid leukaemia (AML) is a cancer of the haematopoietic system, which can in many cases only be cured by haematopoietic stem cell transplantation (HSCT) and donor lymphocyte infusion (DLI) (Burnett et al., 2011). This therapy is associated with the beneficial graft-versus-leukaemia (GvL) effect mediated by transplanted donor T and NK cells that either recognise mismatch HLA molecules or polymorphic peptides, so-called minor histocompatibility antigens, leukaemia-associated or leukaemia-specific antigens in the patient and thus eliminate remaining leukaemic blasts. Nevertheless, the mature donor-derived cells often trigger graft-versus-host disease (GvHD), leading to severe damages in patients’ epithelial tissue, mainly skin, liver and intestine (Bleakley & Riddell, 2004). Therefore, approaches for the selective mediation of strong GvL effects are needed, also in order to prevent relapse after transplantation. One promising opportunity is the in vitro generation of AML-reactive CD4+ T cells for adoptive transfer. CD4+ T cells are advantageous compared to CD8+ T cells, as HLA class II molecules are under non-inflammatory conditions only expressed on haematopoietic cells; a fact that would minimise GvHD (Klein & Sato, 2000). In this study, naive CD4+ T cells were isolated from healthy donors and were successfully stimulated against primary AML blasts in mini-mixed lymphocyte/leukaemia cell cultures (mini-MLLC) in eight patient/donor pairs. After three to seven weekly restimulations, T cells were shown to produce TH1 type cytokines and to be partially of monoclonal origin according to their TCR Vβ chain usage. Furthermore, they exhibited lytic activity towards AML blasts, which was mediated by the release of granzymes A and B and perforin. The patient/donor pairs used in this study were fully HLA-class I matched, except for one pair, and also matched for HLA-DR and -DQ, whereas -DP was mismatched in one or both alleles, reflecting the actual donor selection procedure in the clinic (Begovich et al., 1992). Antibody blocking experiments suggested that the generated CD4+ T cells were directed against the HLA-DP mismatches, which could be confirmed by the recognition of donor-derived lymphoblastoid cell lines (LCLs) electroporated with the mismatched DP alleles. Under non-inflammatory conditions primary fibroblasts did not express HLA-DP and were thus not recognised, supporting the idea of a safer application of CD4+ T cells regarding induction of GvHD. For the assessment of the biological significance of these T cells, they were adoptively transferred into NSG mice engrafted with human AML blasts, where they migrated to the bone marrow and lymphoid tissue and succeeded in eliminating the leukaemic burden after only one week. Therefore, AML-reactive CD4+ T cells expanded from the naive compartment by in vitro stimulation with primary leukaemia blasts appear to be a potent tool for DLI in HSCT patients and promise to mediate specific GvL effects without causing GvHD.

Amphotericin B nano-formulations : development, characterisation and suitability for oral administration

In der Form von Nanokapseln (AmB-HST), Nanoemulsion beziehungsweise multilamellaren Vesikeln (MLV) wurden drei Amphotericin-B-Formulierungen für die orale Applikation entwickelt, charakterisiert und verglichen. Die neuartige homogene Nanokapsel-Formulierung des hydrophoben Polyen-Antimykotikums Amphotericin B wurde in Analogie zu einem für Simvastatin und andere Arzneistoffe etablierten Prozess aus der Reinsubstanz, Lezithin und Gelatine mit Hilfe des HST-Verfahrens hergestellt. Photometrische Untersuchungen zeigten, dass das Endprodukt aus Monomeren aufgebaut ist. Mittels Mikroskopie ließen sich die Aggregate vor der Umhüllung mit Lezithin und Gelatine im Ausgangsmaterial als individuelle kugelförmige Arzneistoffpartikel darstellen. Strukturuntersuchungen mit dynamischer licht streuung (DLS) zeigten eine enge Größenverteilung der verkapselten Partikel von ca. 1 µm. Die Struktur der Hülle der HST-Partikel wurde erstmalig mit Neutronenstreuung unter Verwendung der Deuterium-basierten Lösungsmittel kontrastmethode aufgeklärt. Durch die teilweise Kontrastmaskierung des Partikelkerns bei der Neutronenstreuung konnte die Lezithin-Gelatine-Hülle als eine dünne, 5,64 ± 0.18 nm dicke Schicht aufgelöst werden, welche der biologischen Lipidmembran ähnlich, im Vergleich aber geringfügig größer ist. Dieses Resultat eröffnet Wege für die Optimierung der Formulierung von pharmazeutischen Nanopartikeln, z.B. durch Oberflächenmodifizierungen. Weitere Untersuchungen mittels Kleinwinkelneutronenstreuung unter Verwendung der D-Kontrastvariation deuten darauf hin, dass die Komponenten der Nanokapseln nicht den gleichen Masseschwerpunkt haben, sondern asymmetrisch aufgebaut sind und dass die stärker streuenden Domänen weiter außen liegen. Die Partikel sind im Vergleich zu Liposomen dichter. In-Vitro Freisetzungsstudien belegen das Solubilisierungsvermögen des HST-Systems, wonach die Freisetzung des Arzneistoffes aus der Formulierung zu allen gemessenen Zeitpunkten höher als diejenige der Reinsubstanz war. rnDie Nanoemulsion-Formulierung von Amphotericin B wurde mit einem Öl und Tensid system, jedoch mit unterschiedlichen Co-Solvenzien, erfolgreich entwickelt. Gemäß der Bestimmung der Löslichkeit in verschiedenen Hilfsstoffen erwies sich der Arzneistoff Amphotericin B als nicht-lipophil, gleichzeitig aber auch als nicht-hydrophil. Die zur Ermittlung der für die Emulsionsbildung notwendigen Hilfstoffkonzentrationen erstellten ternären Diagramme veranschaulichten, dass hohe Öl- und Tensidgehalte zu keiner Emulsionsbildung führten. Dementsprechend betrug der höchste Ölgehalt 10%. Die Tröpfchengröße wuchs mit zunehmender Tensidkonzentration, wobei die Co-Solventmenge der Propylenglykol-haltigen Nanoemulsion indirekt verringert wurde. Für die Transcutol®P-haltige Nanoemulsion hingegen wurde das Gegenteil beobachtet, nämlich eine Abnahme der Tröpfchengröße bei steigenden Tensidkonzentrationen. Durch den Einschluss des Arzneistoffes wurde nicht die Viskosität der Formulierung, sondern die Tröpfchengröße beeinflusst. Der Wirkstoffeinschluss führte zu höheren Tröpfchengrößen. Mit zunehmender Propylenglykolkonzentration wurde der Wirkstoffgehalt erhöht, mit zunehmender Transcutol®P-Konzentration dagegen vermindert. UV/VIS-spektroskopische Analysen deuten darauf hin, dass in beiden Formulierungen Amphotericin B als Monomer vorliegt. Allerdings erwiesen sich die Formulierungen Caco-2-Zellen und humanen roten Blutkörperchen gegenüber als toxisch. Da die Kontrollproben eine höhere Toxizität als die wirkstoffhaltigen Formulierungen zeigten, ist die Toxizität nicht nur auf Amphotericin, sondern auch auf die Hilfsstoffe zurückzuführen. Die solubilisierte Wirkstoffmenge ist in beiden Formulierungen nicht ausreichend im Hinblick auf die eingesetzte Menge an Hilfsstoff nach WHO-Kriterien. Gemäß diesen Untersuchungen erscheinen die Emulsions-Formulierungen für die orale Gabe nicht geeignet. Dennoch sind Tierstudien notwendig, um den Effekt bei Tieren sowie die systemisch verfügbare Wirkstoffmenge zu ermitteln. Dies wird bestandskräftige Schlussfolgerungen bezüglich der Formulierung und Aussagen über mögliche Perspektiven erlauben. Nichtsdestotrotz sind die Präkonzentrate sehr stabil und können bei Raumtemperatur gelagert werden.rnDie multilamellar-vesikulären Formulierungen von Amphotericin B mit ungesättigten und gesättigten neutralen Phospholipiden und Cholesterin wurden erfolgreich entwickelt und enthielten nicht nur Vesikel, sondern auch zusätzliche Strukturen bei zunehmender Cholesterinkonzentration. Mittels Partikelgrößenanalyse wurden bei den Formulierungen mit gesättigten Lipiden Mikropartikel detektiert, was abhängig von der Alkylkettenlänge war. Mit dem ungesättigten Lipid (DOPC) konnten hingegen Nanopartikel mit hinreichender Verkapselung und Partikelgrößenverteilung gebildet werden. Die Ergebnisse der thermischen und FTIR-spektroskopischen Analyse, welche den Einfluss des Arzneistoffes ausschließen ließen, liefern den Nachweis für die mögliche, bereits in der Literatur beschriebene Einlagerung des Wirkstoffs in lipid- und/oder cholesterinreiche Membranen. Mit Hilfe eines linearen Saccharosedichtegradienten konnte die Formulierung in Vesikel und Wirkstoff-Lipid-Komplexe nach bimodaler Verteilung aufgetrennt werden, wobei der Arzneistoff stärker mit den Komplexen als mit den Vesikeln assoziiert ist. Bei den Kleinwinkelneutronenstreu-Experimenten wurde die Methode der Kontrastvariation mit Erfolg angewendet. Dabei konnte gezeigt werden, dass Cholesterol in situ einen Komplex mit Amphotericin B bildet. Diesen Sachverhalt legt unter anderem die beobachtete Differenz in der äquivalenten Streulängendichte der Wirkstoff-Lipid- und Wirkstoff-Lipid-Cholesterin-haltigen kleinen unilamellaren Vesikeln nahe. Das Vorkommen von Bragg-Peaks im Streuprofil weist auf Domänen hin und systematische Untersuchungen zeigten, dass die Anzahl der Domänen mit steigendem Cholesteringehalt zunimmt, ab einem bestimmten Grenzwert jedoch wieder abnimmt. Die Domänen treten vor allem nahe der Außenfläche der Modellmembran auf und bestätigen, dass der Wirkstoff in den Cholesterinreichen Membranen vertikal eingelagert ist. Die Formulierung war sowohl Caco-2-Zellen als auch humanen roten Blutkörperchen gegenüber nicht toxisch und erwies sich unter Berücksichtigung der Aufnahme in Caco-2-Zellen als vielversprechend für die orale Applikation. Die Formulierung zeigt sich somit aussichtsreich und könnte in Tabletten weiterverarbeitet werden. Ein Filmüberzug würde den Wirkstoff gegen die saure Umgebung im Magen schützen. Für die Bestimmung der systemischen Verfügbarkeit der Formulierung sind Tierstudien notwendig. Die entwickelten multilamellaren Formulierungen einschließlich der Wirkstoff-Cholesterin-Komplexe bieten somit gute Aussichten auf die mögliche medizinische Anwendung. rnrn

Prediction and measurement of forced response of a composite blade undergoing nonlinear vibration

The main objective of this project is to experimentally demonstrate geometrical nonlinear phenomena due to large displacements during resonant vibration of composite materials and to explain the problem associated with fatigue prediction at resonant conditions. Three different composite blades to be tested were designed and manufactured, being their difference in the composite layup (i.e. unidirectional, cross-ply, and angle-ply layups). Manual envelope bagging technique is explained as applied to the actual manufacturing of the components; problems encountered and their solutions are detailed. Forced response tests of the first flexural, first torsional, and second flexural modes were performed by means of a uniquely contactless excitation system which induced vibration by using a pulsed airflow. Vibration intensity was acquired by means of Polytec LDV system. The first flexural mode is found to be completely linear irrespective of the vibration amplitude. The first torsional mode exhibits a general nonlinear softening behaviour which is interestingly coupled with a hardening behaviour for the unidirectional layup. The second flexural mode has a hardening nonlinear behaviour for either the unidirectional and angle-ply blade, whereas it is slightly softening for the cross-ply layup. By using the same equipment as that used for forced response analyses, free decay tests were performed at different airflow intensities. Discrete Fourier Trasform over the entire decay and Sliding DFT were computed so as to visualise the presence of nonlinear superharmonics in the decay signal and when they were damped out from the vibration over the decay time. Linear modes exhibit an exponential decay, while nonlinearities are associated with a dry-friction damping phenomenon which tends to increase with increasing amplitude. Damping ratio is derived from logarithmic decrement for the exponential branch of the decay.

Dynamic characterisation of four nine-story large-panel R.C. buildings in Bishkek (Kyrgyzstan): A comparison between experimental ambient vibration analysis and numerical finite element modeling

With the outlook of improving seismic vulnerability assessment for the city of Bishkek (Kyrgyzstan), the global dynamic behaviour of four nine-storey r.c. large-panel buildings in elastic regime is studied. The four buildings were built during the Soviet era within a serial production system. Since they all belong to the same series, they have very similar geometries both in plan and in height. Firstly, ambient vibration measurements are performed in the four buildings. The data analysis composed of discrete Fourier transform, modal analysis (frequency domain decomposition) and deconvolution interferometry, yields the modal characteristics and an estimate of the linear impulse response function for the structures of the four buildings. Then, finite element models are set up for all four buildings and the results of the numerical modal analysis are compared with the experimental ones. The numerical models are finally calibrated considering the first three global modes and their results match the experimental ones with an error of less then 20%.

Characterisation of extreme winter precipitation in Mediterranean coastal sites and associated anomalous atmospheric circulation patterns

We present an analysis of daily extreme precipitation events for the extended winter season (October–March) at 20 Mediterranean coastal sites covering the period 1950–2006. The heavy tailed behaviour of precipitation extremes and estimated return levels, including associated uncertainties, are derived applying a procedure based on the Generalized Pareto Distribution, in combination with recently developed methods. Precipitation extremes have an important contribution to make seasonal totals (approximately 60% for all series). Three stations (one in the western Mediterranean and the others in the eastern basin) have a 5-year return level above 100 mm, while the lowest value (estimated for two Italian series) is equal to 58 mm. As for the 50-year return level, an Italian station (Genoa) has the highest value of 264 mm, while the other values range from 82 to 200 mm. Furthermore, six series (from stations located in France, Italy, Greece, and Cyprus) show a significant negative tendency in the probability of observing an extreme event. The relationship between extreme precipitation events and the large scale atmospheric circulation at the upper, mid and low troposphere is investigated by using NCEP/NCAR reanalysis data. A 2-step classification procedure identifies three significant anomaly patterns both for the western-central and eastern part of the Mediterranean basin. In the western Mediterranean, the anomalous southwesterly surface to mid-tropospheric flow is connected with enhanced moisture transport from the Atlantic. During ≥5-year return level events, the subtropical jet stream axis is aligned with the African coastline and interacts with the eddy-driven jet stream. This is connected with enhanced large scale ascending motions, instability and leads to the development of severe precipitation events. For the eastern Mediterranean extreme precipitation events, the identified anomaly patterns suggest warm air advection connected with anomalous ascent motions and an increase of the low- to mid-tropospheric moisture. Furthermore, the jet stream position (during ≥5-year return level events) supports the eastern basin being in a divergence area, where ascent motions are favoured. Our results contribute to an improved understanding of daily precipitation extremes in the cold season and associated large scale atmospheric features.

Dynamic modeling, simulation and control design of a parafoil-payload system for ship launched aerial delivery system (SLADS)

The objective of this research was to develop a high-fidelity dynamic model of a parafoilpayload system with respect to its application for the Ship Launched Aerial Delivery System (SLADS). SLADS is a concept in which cargo can be transfered from ship to shore using a parafoil-payload system. It is accomplished in two phases: An initial towing phase when the glider follows the towing vessel in a passive lift mode and an autonomous gliding phase when the system is guided to the desired point. While many previous researchers have analyzed the parafoil-payload system when it is released from another airborne vehicle, limited work has been done in the area of towing up the system from ground or sea. One of the main contributions of this research was the development of a nonlinear dynamic model of a towed parafoil-payload system. After performing an extensive literature review of the existing methods of modeling a parafoil-payload system, a five degree-of-freedom model was developed. The inertial and geometric properties of the system were investigated to predict accurate results in the simulation environment. Since extensive research has been done in determining the aerodynamic characteristics of a paraglider, an existing aerodynamic model was chosen to incorporate the effects of air flow around the flexible paraglider wing. During the towing phase, it is essential that the parafoil-payload system follow the line of the towing vessel path to prevent an unstable flight condition called ‘lockout’. A detailed study of the causes of lockout, its mathematical representation and the flight conditions and the parameters related to lockout, constitute another contribution of this work. A linearized model of the parafoil-payload system was developed and used to analyze the stability of the system about equilibrium conditions. The relationship between the control surface inputs and the stability was investigated. In addition to stability of flight, one more important objective of SLADS is to tow up the parafoil-payload system as fast as possible. The tension in the tow cable is directly proportional to the rate of ascent of the parafoil-payload system. Lockout instability is more favorable when tow tensions are large. Thus there is a tradeoff between susceptibility to lockout and rapid deployment. Control strategies were also developed for optimal tow up and to maintain stability in the event of disturbances.

Non-invasive method to predict viscosity and size using total internal reflection fluorescence microscopy (TIRFM) system

This study develops an automated analysis tool by combining total internal reflection fluorescence microscopy (TIRFM), an evanescent wave microscopic imaging technique to capture time-sequential images and the corresponding image processing Matlab code to identify movements of single individual particles. The developed code will enable us to examine two dimensional hindered tangential Brownian motion of nanoparticles with a sub-pixel resolution (nanoscale). The measured mean square displacements of nanoparticles are compared with theoretical predictions to estimate particle diameters and fluid viscosity using a nonlinear regression technique. These estimated values will be confirmed by the diameters and viscosities given by manufacturers to validate this analysis tool. Nano-particles used in these experiments are yellow-green polystyrene fluorescent nanospheres (200 nm, 500 nm and 1000 nm in diameter (nominal); 505 nm excitation and 515 nm emission wavelengths). Solutions used in this experiment are de-ionized (DI) water, 10% d-glucose and 10% glycerol. Mean square displacements obtained near the surface shows significant deviation from theoretical predictions which are attributed to DLVO forces in the region but it conforms to theoretical predictions after ~125 nm onwards. The proposed automation analysis tool will be powerfully employed in the bio-application fields needed for examination of single protein (DNA and/or vesicle) tracking, drug delivery, and cyto-toxicity unlike the traditional measurement techniques that require fixing the cells. Furthermore, this tool can be also usefully applied for the microfluidic areas of non-invasive thermometry, particle tracking velocimetry (PTV), and non-invasive viscometry.

Gaze behavior nonlinear dynamics assessed in virtual immersion as a diagnostic index of sexual deviancy: preliminary results

This paper presents preliminary results about the use of virtual characters, penile plethysmography and gaze behaviour dynamics to assess deviant sexual preferences. Pedophile patients’ responses are compared to those of non-deviant subjects while they were immersed with virtual characters depicting relevant sexual features.

Extended kalman filter for estimation of parameters in nonlinear state-space models of biochemical networks.

It is system dynamics that determines the function of cells, tissues and organisms. To develop mathematical models and estimate their parameters are an essential issue for studying dynamic behaviors of biological systems which include metabolic networks, genetic regulatory networks and signal transduction pathways, under perturbation of external stimuli. In general, biological dynamic systems are partially observed. Therefore, a natural way to model dynamic biological systems is to employ nonlinear state-space equations. Although statistical methods for parameter estimation of linear models in biological dynamic systems have been developed intensively in the recent years, the estimation of both states and parameters of nonlinear dynamic systems remains a challenging task. In this report, we apply extended Kalman Filter (EKF) to the estimation of both states and parameters of nonlinear state-space models. To evaluate the performance of the EKF for parameter estimation, we apply the EKF to a simulation dataset and two real datasets: JAK-STAT signal transduction pathway and Ras/Raf/MEK/ERK signaling transduction pathways datasets. The preliminary results show that EKF can accurately estimate the parameters and predict states in nonlinear state-space equations for modeling dynamic biochemical networks.

A system design for space-based space surveillance

This paper presents the capabilities of a Space-Based Space Surveillance (SBSS) demonstration mission for Space Surveillance and Tracking (SST) based on a micro- satellite platform. The results have been produced in the frame of ESA’s "As sessment Study for Space Based Space Surveillance Demonstration Mission (Phase A) " performed by the Airbus DS consortium. Space Surveillance and Tracking is part of Space Situational Awareness (SSA) and covers the detection, tracking and cataloguing of spa ce debris and satellites. Derived SST services comprise a catalogue of these man-made objects, collision warning, detection and characterisation of in-orbit fragmentations, sub-catalogue debris characterisation, etc. The assessment of SBSS in an SST system architecture has shown that both an operational SBSS and also already a well - designed space-based demonstrator can provide substantial performance in terms of surveillance and tracking of beyond - LEO objects. Especially the early deployment of a demonstrator, possible by using standard equipment, could boost initial operating capability and create a self-maintained object catalogue. Unlike classical technology demonstration missions, the primary goal is the demonstration and optimisation of the functional elements in a complex end-to-end chain (mission planning, observation strategies, data acquisition, processing and fusion, etc.) until the final products can be offered to the users. The presented SBSS system concept takes the ESA SST System Requirements (derived within the ESA SSA Preparatory Program) into account and aims at fulfilling some of the SST core requirements in a stand-alone manner. The evaluation of the concept has shown that an according solution can be implemented with low technological effort and risk. The paper presents details of the system concept, candidate micro - satellite platforms, the observation strategy and the results of performance simulations for GEO coverage and cataloguing accuracy

Space-based space surveillance and tracking demonstrator: mission and system design

This paper presents the capabilities of a Space-Based Space Surveillance (SBSS) demonstration mission for Space Surveillance and Tracking (SST) based on a micro-satellite platform. The results have been produced in the frame of ESA’s "Assessment Study for Space Based Space Surveillance Demonstration Mission" performed by the Airbus Defence and Space consortium. The assessment of SBSS in an SST system architecture has shown that both an operational SBSS and also already a well- designed space-based demonstrator can provide substantial performance in terms of surveillance and tracking of beyond-LEO objects. Especially the early deployment of a demonstrator, possible by using standard equipment, could boost initial operating capability and create a self-maintained object catalogue. Furthermore, unique statistical information about small-size LEO debris (mm size) can be collected in-situ. Unlike classical technology demonstration missions, the primary goal is the demonstration and optimisation of the functional elements in a complex end-to-end chain (mission planning, observation strategies, data acquisition, processing, etc.) until the final products can be offered to the users and with low technological effort and risk. The SBSS system concept takes the ESA SST System Requirements into account and aims at fulfilling SST core requirements in a stand-alone manner. Additionally, requirements for detection and characterisation of small-sizedLEO debris are considered. The paper presents details of the system concept, candidate micro-satellite platforms, the instrument design and the operational modes. Note that the detailed results of performance simulations for space debris coverage and cataloguing accuracy are presented in a separate paper “Capability of a Space-based Space Surveillance System to Detect and Track Objects in GEO, MEO and LEO Orbits” by J. Silha (AIUB) et al., IAC-14, A6, 1.1x25640.

A Passive Source Localization System for IEEE 802.15.4 Signal

In this work, we provide a passive location monitoring system for IEEE 802.15.4 signal emitters. The system adopts software defined radio techniques to passively overhear IEEE 802.15.4 packets and to extract power information from baseband signals. In our system, we provide a new model based on the nonlinear regression for ranging. After obtaining distance information, a Weighted Centroid (WC) algorithm is adopted to locate users. In WC, each weight is inversely proportional to the nth power of propagation distance, and the degree n is obtained from some initial measurements. We evaluate our system in a 16m-18m area with complex indoor propagation conditions. We are able to achieve a median error of 2:1m with only 4 anchor nodes.

A Passive WiFi Source Localization System based on Fine-grained Power-based Trilateration

Indoor localization systems become more interesting for researchers because of the attractiveness of business cases in various application fields. A WiFi-based passive localization system can provide user location information to third-party providers of positioning services. However, indoor localization techniques are prone to multipath and Non-Line Of Sight (NLOS) propagation, which lead to significant performance degradation. To overcome these problems, we provide a passive localization system for WiFi targets with several improved algorithms for localization. Through Software Defined Radio (SDR) techniques, we extract Channel Impulse Response (CIR) information at the physical layer. CIR is later adopted to mitigate the multipath fading problem. We propose to use a Nonlinear Regression (NLR) method to relate the filtered power information to propagation distances, which significantly improves the ranging accuracy compared to the commonly used log-distance path loss model. To mitigate the influence of ranging errors, a new trilateration algorithm is designed as well by combining Weighted Centroid and Constrained Weighted Least Square (WC-CWLS) algorithms. Experiment results show that our algorithm is robust against ranging errors and outperforms the linear least square algorithm and weighted centroid algorithm.

A characterisation of passive and active interactions and their influence on student's achievement using Moodle LMS Logs

During the last years, there has been much concern about learning management systems' (LMS) effectiveness when compared to traditional learning and about how to assess students' participation during the course. The tracking and monitoring capabilities of most recent LMS have made it possible to analyse every interaction in the system. The issues addressed on this study are: a) Is LMS student's interaction an indicator of academic performance?; b) Are different results in performance expected between distance and in-class LMS-supported education?; c) How can LMS interactions from logs be categorised?; d) May this categorisation detect 'learning witnesses'? To answer these questions, a set of interaction types from Moodle LMS activity record logs has been analysed during two years in online and in-class Master's degrees at the UPM. The results show partial or no evidence of influence between interaction indicators and academic performance, although the proposed categorisation may help detect learning witnesses.

A new approach to fuzzy estimation of Takagi-Sugeno model and its applications to optimal control for nonlinear systems

An efficient approach is presented to improve the local and global approximation and modelling capability of Takagi-Sugeno (T-S) fuzzy model. The main aim is obtaining high function approximation accuracy. The main problem is that T-S identification method cannot be applied when the membership functions are overlapped by pairs. This restricts the use of the T-S method because this type of membership function has been widely used during the last two decades in the stability, controller design and are popular in industrial control applications. The approach developed here can be considered as a generalized version of T-S method with optimized performance in approximating nonlinear functions. A simple approach with few computational effort, based on the well known parameters' weighting method is suggested for tuning T-S parameters to improve the choice of the performance index and minimize it. A global fuzzy controller (FC) based Linear Quadratic Regulator (LQR) is proposed in order to show the effectiveness of the estimation method developed here in control applications. Illustrative examples of an inverted pendulum and Van der Pol system are chosen to evaluate the robustness and remarkable performance of the proposed method and the high accuracy obtained in approximating nonlinear and unstable systems locally and globally in comparison with the original T-S model. Simulation results indicate the potential, simplicity and generality of the algorithm.