Modeling and simulation of a synthetic genetic circuit that implements a multicelled behavior in a growing microcolony of E. coli

Synthetic Biology is a relatively new discipline, born at the beginning of the New Millennium, that brings the typical engineering approach (abstraction, modularity and standardization) to biotechnology. These principles aim to tame the extreme complexity of the various components and aid the construction of artificial biological systems with specific functions, usually by means of synthetic genetic circuits implemented in bacteria or simple eukaryotes like yeast. The cell becomes a programmable machine and its low-level programming language is made of strings of DNA. This work was performed in collaboration with researchers of the Department of Electrical Engineering of the University of Washington in Seattle and also with a student of the Corso di Laurea Magistrale in Ingegneria Biomedica at the University of Bologna: Marilisa Cortesi. During the collaboration I contributed to a Synthetic Biology project already started in the Klavins Laboratory. In particular, I modeled and subsequently simulated a synthetic genetic circuit that was ideated for the implementation of a multicelled behavior in a growing bacterial microcolony. In the first chapter the foundations of molecular biology are introduced: structure of the nucleic acids, transcription, translation and methods to regulate gene expression. An introduction to Synthetic Biology completes the section. In the second chapter is described the synthetic genetic circuit that was conceived to make spontaneously emerge, from an isogenic microcolony of bacteria, two different groups of cells, termed leaders and followers. The circuit exploits the intrinsic stochasticity of gene expression and intercellular communication via small molecules to break the symmetry in the phenotype of the microcolony. The four modules of the circuit (coin flipper, sender, receiver and follower) and their interactions are then illustrated. In the third chapter is derived the mathematical representation of the various components of the circuit and the several simplifying assumptions are made explicit. Transcription and translation are modeled as a single step and gene expression is function of the intracellular concentration of the various transcription factors that act on the different promoters of the circuit. A list of the various parameters and a justification for their value closes the chapter. In the fourth chapter are described the main characteristics of the gro simulation environment, developed by the Self Organizing Systems Laboratory of the University of Washington. Then, a sensitivity analysis performed to pinpoint the desirable characteristics of the various genetic components is detailed. The sensitivity analysis makes use of a cost function that is based on the fraction of cells in each one of the different possible states at the end of the simulation and the wanted outcome. Thanks to a particular kind of scatter plot, the parameters are ranked. Starting from an initial condition in which all the parameters assume their nominal value, the ranking suggest which parameter to tune in order to reach the goal. Obtaining a microcolony in which almost all the cells are in the follower state and only a few in the leader state seems to be the most difficult task. A small number of leader cells struggle to produce enough signal to turn the rest of the microcolony in the follower state. It is possible to obtain a microcolony in which the majority of cells are followers by increasing as much as possible the production of signal. Reaching the goal of a microcolony that is split in half between leaders and followers is comparatively easy. The best strategy seems to be increasing slightly the production of the enzyme. To end up with a majority of leaders, instead, it is advisable to increase the basal expression of the coin flipper module. At the end of the chapter, a possible future application of the leader election circuit, the spontaneous formation of spatial patterns in a microcolony, is modeled with the finite state machine formalism. The gro simulations provide insights into the genetic components that are needed to implement the behavior. In particular, since both the examples of pattern formation rely on a local version of Leader Election, a short-range communication system is essential. Moreover, new synthetic components that allow to reliably downregulate the growth rate in specific cells without side effects need to be developed. In the appendix are listed the gro code utilized to simulate the model of the circuit, a script in the Python programming language that was used to split the simulations on a Linux cluster and the Matlab code developed to analyze the data.

Multivariate analysis methods in the search for the Higgs boson produced in association with top pairs at the ATLAS experiment at LHC.

The Large Hadron Collider, located at the CERN laboratories in Geneva, is the largest particle accelerator in the world. One of the main research fields at LHC is the study of the Higgs boson, the latest particle discovered at the ATLAS and CMS experiments. Due to the small production cross section for the Higgs boson, only a substantial statistics can offer the chance to study this particle properties. In order to perform these searches it is desirable to avoid the contamination of the signal signature by the number and variety of the background processes produced in pp collisions at LHC. Much account assumes the study of multivariate methods which, compared to the standard cut-based analysis, can enhance the signal selection of a Higgs boson produced in association with a top quark pair through a dileptonic final state (ttH channel). The statistics collected up to 2012 is not sufficient to supply a significant number of ttH events; however, the methods applied in this thesis will provide a powerful tool for the increasing statistics that will be collected during the next LHC data taking.

Small polarons in spin-orbit coupled osmates

Small polarons (SP) have been thoroughly investigated in 3d transition metal oxides and they have been found to play a crucial role in physical phenomena such as charge transport, colossal magnetoresistance and surface reactivity. However, our knowledge about these quasi-particles in 5d systems remains very limited, since the more delocalised nature of the 5d orbitals reduces the strength of the Electronic Correlation (EC), making SP formation in these compounds rather unexpected. Nevertheless, the Spin-Orbit coupled Dirac-Mott insulator Ba2NaOsO6 (BNOO) represents a good candidate for enabling polaron formation in a relativistic background, due to the relatively large EC (U ∼ 3 eV) and Jahn-Teller activity. Moreover, anomalous peaks in Nuclear Magnetic Resonance (NMR) spectroscopy experiments suggest the presence of thermally activated SP dynamics when BNOO is doped with Ca atoms. We investigate SP formation in BNOO both from an electronic and structural point of view by means of fully relativistic first principles calculations. Our numerical simulations predict a stable SP ground state and agree on the value of 810 K for the dynamical process peak found by NMR experiments.

Data-driven Signal Processing per Jamming Detection in Sistemi Satellitari

In questo elaborato vengono analizzate differenti tecniche per la detection di jammer attivi e costanti in una comunicazione satellitare in uplink. Osservando un numero limitato di campioni ricevuti si vuole identificare la presenza di un jammer. A tal fine sono stati implementati i seguenti classificatori binari: support vector machine (SVM), multilayer perceptron (MLP), spectrum guarding e autoencoder. Questi algoritmi di apprendimento automatico dipendono dalle features che ricevono in ingresso, per questo motivo è stata posta particolare attenzione alla loro scelta. A tal fine, sono state confrontate le accuratezze ottenute dai detector addestrati utilizzando differenti tipologie di informazione come: i segnali grezzi nel tempo, le statistical features, le trasformate wavelet e lo spettro ciclico. I pattern prodotti dall’estrazione di queste features dai segnali satellitari possono avere dimensioni elevate, quindi, prima della detection, vengono utilizzati i seguenti algoritmi per la riduzione della dimensionalità: principal component analysis (PCA) e linear discriminant analysis (LDA). Lo scopo di tale processo non è quello di eliminare le features meno rilevanti, ma combinarle in modo da preservare al massimo l’informazione, evitando problemi di overfitting e underfitting. Le simulazioni numeriche effettuate hanno evidenziato come lo spettro ciclico sia in grado di fornire le features migliori per la detection producendo però pattern di dimensioni elevate, per questo motivo è stato necessario l’utilizzo di algoritmi di riduzione della dimensionalità. In particolare, l'algoritmo PCA è stato in grado di estrarre delle informazioni migliori rispetto a LDA, le cui accuratezze risentivano troppo del tipo di jammer utilizzato nella fase di addestramento. Infine, l’algoritmo che ha fornito le prestazioni migliori è stato il Multilayer Perceptron che ha richiesto tempi di addestramento contenuti e dei valori di accuratezza elevati.

Development of a state-variable filter for a motion control application.

In collaboration with G.D. SpA I attended an internship with the purpose of developing a filter for the position control of industrial machines during testing and maintenance operations. The filter elaborates a signal in position provided by an electonic handwheel, in order to enable the application to be controlled with a signal in velocity with arbitrarily dynamics chosen during the design phase. Limiting the dynamics of the filter provide a more stable and less demanding reference trajectory which reduce the vibrations and tracking errors of the motor controlled by it. It also prevents misusages of the handwheel from the technician which could end up in harmful interferences between the mechanical parts moved by the handwheel.