A proteo-liposome system for the analysis of the intracellular interactome of membrane proteins using amyloid precursor protein as a model

Transmembrane proteins play crucial roles in many important physiological processes. The intracellular domain of membrane proteins is key for their function by interacting with a wide variety of cytosolic proteins. It is therefore important to examine this interaction. A recently developed method to study these interactions, based on the use of liposomes as a model membrane, involves the covalent coupling of the cytoplasmic domains of membrane proteins to the liposome membrane. This allows for the analysis of interaction partners requiring both protein and membrane lipid binding. This thesis further establishes the liposome recruitment system and utilises it to examine the intracellular interactome of the amyloid precursor protein (APP), most well-known for its proteolytic cleavage that results in the production and accumulation of amyloid beta fragments, the main constituent of amyloid plaques in Alzheimer’s disease pathology. Despite this, the physiological function of APP remains largely unclear. Through the use of the proteo-liposome recruitment system two novel interactions of APP’s intracellular domain (AICD) are examined with a view to gaining a greater insight into APP’s physiological function. One of these novel interactions is between AICD and the mTOR complex, a serine/threonine protein kinase that integrates signals from nutrients and growth factors. The kinase domain of mTOR directly binds to AICD and the N-terminal amino acids of AICD are crucial for this interaction. The second novel interaction is between AICD and the endosomal PIKfyve complex, a lipid kinase involved in the production of phosphatidylinositol-3,5-bisphosphate (PI(3,5)P2) from phosphatidylinositol-3-phosphate, which has a role in controlling ensdosome dynamics. The scaffold protein Vac14 of the PIKfyve complex binds directly to AICD and the C-terminus of AICD is important for its interaction with the PIKfyve complex. Using a recently developed intracellular PI(3,5)P2 probe it is shown that APP controls the formation of PI(3,5)P2 positive vesicular structures and that the PIKfyve complex is involved in the trafficking and degradation of APP. Both of these novel APP interactors have important implications of both APP function and Alzheimer’s disease. The proteo-liposome recruitment method is further validated through its use to examine the recruitment and assembly of the AP-2/clathrin coat from purified components to two membrane proteins containing different sorting motifs. Taken together this thesis highlights the proteo-liposome recruitment system as a valuable tool for the study of membrane proteins intracellular interactome. It allows for the mimicking of the protein in its native configuration therefore identifying weaker interactions that are not detected by more conventional methods and also detecting interactions that are mediated by membrane phospholipids.

The impact of pump phase-modulation and filtering on WDM signals in a fibre optical parametric amplifier

WDM signal degradation from pump phase-modulation in a one-pump 20dB net-gain fibre optical parametric amplifier is experimentally and numerically characterised for the first time using 10x59Gb/s QPSK signals.

Comparison of software developed for FHR extraction from PCG signals

A valuable alternative to US cardiotocography, for fetal surveillance, can be offered by phonocardiography, a passive and low cost acoustic recording of fetal heart sounds. A crucial point is the exact recognizing of the fetal heart sounds, associated to each fetal heart beat, and then the estimation of FHR signal. In this work, software for FHR assessment from phonocardiographic signals was developed. To check the reliability of the software, obtained results were compared with those of simultaneously recorded cardiotocographic signals. Results seemed to be satisfying, as provided FHR series were almost all confined within FHR-CTG +/- 3 bpm, where FHR-CTG were FHR series provided by commercial US cardiotocographic devices, currently employed in clinical routine.

An algorithm for FHR estimation from foetal phonocardiographic signals

The long-term foetal surveillance is often to be recommended. Hence, the fully non-invasive acoustic recording, through maternal abdomen, represents a valuable alternative to the ultrasonic cardiotocography. Unfortunately, the recorded heart sound signal is heavily loaded by noise, thus the determination of the foetal heart rate raises serious signal processing issues. In this paper, we present a new algorithm for foetal heart rate estimation from foetal phonocardiographic recordings. A filtering is employed as a first step of the algorithm to reduce the background noise. A block for first heart sounds enhancing is then used to further reduce other components of foetal heart sound signals. A complex logic block, guided by a number of rules concerning foetal heart beat regularity, is proposed as a successive block, for the detection of most probable first heart sounds from several candidates. A final block is used for exact first heart sound timing and in turn foetal heart rate estimation. Filtering and enhancing blocks are actually implemented by means of different techniques, so that different processing paths are proposed. Furthermore, a reliability index is introduced to quantify the consistency of the estimated foetal heart rate and, based on statistic parameters; [,] a software quality index is designed to indicate the most reliable analysis procedure (that is, combining the best processing path and the most accurate time mark of the first heart sound, provides the lowest estimation errors). The algorithm performances have been tested on phonocardiographic signals recorded in a local gynaecology private practice from a sample group of about 50 pregnant women. Phonocardiographic signals have been recorded simultaneously to ultrasonic cardiotocographic signals in order to compare the two foetal heart rate series (the one estimated by our algorithm and the other provided by cardiotocographic device). Our results show that the proposed algorithm, in particular some analysis procedures, provides reliable foetal heart rate signals, very close to the reference cardiotocographic recordings. © 2010 Elsevier Ltd. All rights reserved.

A simulating software of fetal phonocardiographic signals

A valuable alternative to traditional diagnostic tool to record fetal heart rate, to monitor the general fetal wellbeing, is fetal phonocardiography, a passive and low cost acoustic recording of fetal heart sounds. In this paper, it is presented a simulating software of fetal phonocardiographic signals relative to different fetal physiological states and recording conditions (for example different kinds and levels of noise). This software can be useful to test and assess fetal heart rate extraction algorithms from fetal phonocardiographic recordings and as a teaching tool for demonstration to medical students and others. © 2010 IEEE.

Fibre impairment compensation using artificial neural network equalizer for high-capacity coherent optical OFDM signals

We propose an artificial neural network (ANN) equalizer for transmission performance enhancement of coherent optical OFDM (C-OOFDM) signals. The ANN equalizer showed more efficiency in combating both chromatic dispersion (CD) and single-mode fibre (SMF)-induced non-linearities compared to the least mean square (LMS). The equalizer can offer a 1.5 dB improvement in optical signal-to-noise ratio (OSNR) compared to LMS algorithm for 40 Gbit/s C-OOFDM signals when considering only CD. It is also revealed that ANN can double the transmission distance up to 320 km of SMF compared to the case of LMS, providing a nonlinearity tolerance improvement of ∼0.7 dB OSNR.

All-optical add-drop multiplexing of OFDM signals

We discuss the recently proposed architecture for an all-optical add-drop multiplexer of OFDM signals and we summarize the results of its theoretical design and experimental implementation. © 2015 OSA.

Direction Finding Estimators of Cyclostationary Signals in Array Processing for Microwave Power Transmission

A solar power satellite is paid attention to as a clean, inexhaustible large- scale base-load power supply. The following technology related to beam control is used: A pilot signal is sent from the power receiving site and after direction of arrival estimation the beam is directed back to the earth by same direction. A novel direction-finding algorithm based on linear prediction technique for exploiting cyclostationary statistical information (spatial and temporal) is explored. Many modulated communication signals exhibit a cyclostationarity (or periodic correlation) property, corresponding to the underlying periodicity arising from carrier frequencies or baud rates. The problem was solved by using both cyclic second-order statistics and cyclic higher-order statistics. By evaluating the corresponding cyclic statistics of the received data at certain cycle frequencies, we can extract the cyclic correlations of only signals with the same cycle frequency and null out the cyclic correlations of stationary additive noise and all other co-channel interferences with different cycle frequencies. Thus, the signal detection capability can be significantly improved. The proposed algorithms employ cyclic higher-order statistics of the array output and suppress additive Gaussian noise of unknown spectral content, even when the noise shares common cycle frequencies with the non-Gaussian signals of interest. The proposed method completely exploits temporal information (multiple lag ), and also can correctly estimate direction of arrival of desired signals by suppressing undesired signals. Our approach was generalized over direction of arrival estimation of cyclostationary coherent signals. In this paper, we propose a new approach for exploiting cyclostationarity that seems to be more advanced in comparison with the other existing direction finding algorithms.

All-optical add-drop multiplexer for OFDM signals

We summarize our research work on the design and development of an add-drop multiplexer for spectrally overlapping OFDM signals. The standard node functions of sub-channel drop, extraction and insertion were obtained whilst the signals remained fully in the optical domain. Numerical simulations have been carried out to identify the main sources of degradation and to benchmark the architectural performance against critical design parameters, whereas the experimental demonstration of the scheme has been achieved for both single quadrature and dual quadrature signals. The reported scheme enables a fully flexible node compatible with future terabit per second super-channel transmission.

Accent Recognition for Noisy Audio Signals

It is well established that accent recognition can be as accurate as up to 95% when the signals are noise-free, using feature extraction techniques such as mel-frequency cepstral coefficients and binary classifiers such as discriminant analysis, support vector machine and k-nearest neighbors. In this paper, we demonstrate that the predictive performance can be reduced by as much as 15% when the signals are noisy. Specifically, in this paper we perturb the signals with different levels of white noise, and as the noise become stronger, the out-of-sample predictive performance deteriorates from 95% to 80%, although the in-sample prediction gives overly-optimistic results. ACM Computing Classification System (1998): C.3, C.5.1, H.1.2, H.2.4., G.3.

Numerical investigation of all-optical add-drop multiplexing for spectrally overlapping OFDM signals

We propose a novel architecture for all-optical add-drop multiplexing of OFDM signals. Sub-channel extraction is achieved by means of waveform replication and coherent subtraction from the OFDM super-channel. Numerical simulations have been carried out to benchmark the performance of the architecture against critical design parameters.

Two cascaded SOAs used as intensity modulators for adaptively modulated optical OFDM signals in optical access networks

Detailed theoretical and numerical investigations of the transmission performance of adaptively modulated optical orthogonal frequency division multiplexed (AMOOFDM) signals are undertaken, for the first time, in optical amplification and chromatic dispersion (CD) compensation free single mode fiber (SMF) intensity-modulated and directdetection (IMDD) systems using two cascaded semiconductor optical amplifiers in a counterpropagating configuration as an intensity modulator (TC-SOA-CC-IM). A theoretical model describing the characteristics of this configuration is developed. Extensive performance comparisons are also made between the TC-SOA-CC and the single SOA intensity modulators. It is shown that, the TC-SOA-CC reaches its strongly saturated region using a lower input optical power much faster than the single SOA resulting in significantly reduced effective carrier lifetime and thus wide TC-SOA-CC bandwidths. It is shown that at low input optical power, we can increase the signal line rate almost 115% which will be more than twice the transmission performance offered by single SOA. In addition, the TCSOA-CC-IM is capable of supporting signal line rates higher than corresponding to the SOA-IM by using 10dB lower input optical powers. For long transmission distance, the TC-SOA-CC-IM has much stronger CD compensation capability compared to the SOA-IM. In addition the use of TC-SOA-CC-IM is more effective regarding the capability to benefit from the CD compensation for shorter distances starting at 60km SMF, whilst for the SOA-IM starting at 90km. © 2014 Optical Society of America.

The development of attenuation compensation models of fluorescence spectroscopy signals

This study examines the effect of blood absorption on the endogenous fluorescence signal intensity of biological tissues. Experimental studies were conducted to identify these effects. To register the fluorescence intensity, the fluorescence spectroscopy method was employed. The intensity of the blood flow was measured by laser Doppler flowmetry. We proposed one possible implementation of the Monte Carlo method for the theoretical analysis of the effect of blood on the fluorescence signals. The simulation is constructed as a four-layer skin optical model based on the known optical parameters of the skin with different levels of blood supply. With the help of the simulation, we demonstrate how the level of blood supply can affect the appearance of the fluorescence spectra. In addition, to describe the properties of biological tissue, which may affect the fluorescence spectra, we turned to the method of diffuse reflectance spectroscopy (DRS). Using the spectral data provided by the DRS, the tissue attenuation effect can be extracted and used to correct the fluorescence spectra.

Teaching Sorting in ICT

This article is aimed at considering how an algorithmic problem - more precisely a sorting problem - can be used in an informatics class in primary and secondary education to make students mobilize the largest possible amount of their intellectual skills in the problem solving process. We will be outlining a method which essentially forces students to utilize their mathematical knowledge besides algorithmization in order to provide an efficient solution. What is more, they are expected to use efficiently a tool that has so far not been associated with creative thinking. Sorting is meant to be just an example, through which our thoughts can easily be demonstrated, but - of course the method of education outlined can be linked to several other algorithmic problems, as well.

Signal processing of physiological signals for automated assessment of stress in computer users

The need to provide computers with the ability to distinguish the affective state of their users is a major requirement for the practical implementation of affective computing concepts. This dissertation proposes the application of signal processing methods on physiological signals to extract from them features that can be processed by learning pattern recognition systems to provide cues about a person's affective state. In particular, combining physiological information sensed from a user's left hand in a non-invasive way with the pupil diameter information from an eye-tracking system may provide a computer with an awareness of its user's affective responses in the course of human-computer interactions. In this study an integrated hardware-software setup was developed to achieve automatic assessment of the affective status of a computer user. A computer-based "Paced Stroop Test" was designed as a stimulus to elicit emotional stress in the subject during the experiment. Four signals: the Galvanic Skin Response (GSR), the Blood Volume Pulse (BVP), the Skin Temperature (ST) and the Pupil Diameter (PD), were monitored and analyzed to differentiate affective states in the user. Several signal processing techniques were applied on the collected signals to extract their most relevant features. These features were analyzed with learning classification systems, to accomplish the affective state identification. Three learning algorithms: Naïve Bayes, Decision Tree and Support Vector Machine were applied to this identification process and their levels of classification accuracy were compared. The results achieved indicate that the physiological signals monitored do, in fact, have a strong correlation with the changes in the emotional states of the experimental subjects. These results also revealed that the inclusion of pupil diameter information significantly improved the performance of the emotion recognition system. ^