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.

Simulation of bistatic radar experiments with deep space missions

L’obiettivo del lavoro esposto nella seguente relazione di tesi ha riguardato lo studio e la simulazione di esperimenti di radar bistatico per missioni di esplorazione planeteria. In particolare, il lavoro si è concentrato sull’uso ed il miglioramento di un simulatore software già realizzato da un consorzio di aziende ed enti di ricerca nell’ambito di uno studio dell’Agenzia Spaziale Europea (European Space Agency – ESA) finanziato nel 2008, e svolto fra il 2009 e 2010. L’azienda spagnola GMV ha coordinato lo studio, al quale presero parte anche gruppi di ricerca dell’Università di Roma “Sapienza” e dell’Università di Bologna. Il lavoro svolto si è incentrato sulla determinazione della causa di alcune inconsistenze negli output relativi alla parte del simulatore, progettato in ambiente MATLAB, finalizzato alla stima delle caratteristiche della superficie di Titano, in particolare la costante dielettrica e la rugosità media della superficie, mediante un esperimento con radar bistatico in modalità downlink eseguito dalla sonda Cassini-Huygens in orbita intorno al Titano stesso. Esperimenti con radar bistatico per lo studio di corpi celesti sono presenti nella storia dell’esplorazione spaziale fin dagli anni ’60, anche se ogni volta le apparecchiature utilizzate e le fasi di missione, durante le quali questi esperimenti erano effettuati, non sono state mai appositamente progettate per lo scopo. Da qui la necessità di progettare un simulatore per studiare varie possibili modalità di esperimenti con radar bistatico in diversi tipi di missione. In una prima fase di approccio al simulatore, il lavoro si è incentrato sullo studio della documentazione in allegato al codice così da avere un’idea generale della sua struttura e funzionamento. È seguita poi una fase di studio dettagliato, determinando lo scopo di ogni linea di codice utilizzata, nonché la verifica in letteratura delle formule e dei modelli utilizzati per la determinazione di diversi parametri. In una seconda fase il lavoro ha previsto l’intervento diretto sul codice con una serie di indagini volte a determinarne la coerenza e l’attendibilità dei risultati. Ogni indagine ha previsto una diminuzione delle ipotesi semplificative imposte al modello utilizzato in modo tale da identificare con maggiore sicurezza la parte del codice responsabile dell’inesattezza degli output del simulatore. I risultati ottenuti hanno permesso la correzione di alcune parti del codice e la determinazione della principale fonte di errore sugli output, circoscrivendo l’oggetto di studio per future indagini mirate.

Development of the Simulation Model for the CoSES Laboratory Test Microgrid in Modelica

Evolution of the traditional consumer in a power system to a prosumer has posed many problems in the traditional uni-directional grid. This evolution in the grid model has made it important to study the behaviour of microgrids. This thesis deals with the laboratory microgrid setup at the Munich School of Engineering, built to assist researchers in studying microgrids. The model is built in Dymola which is a tool for the OpenModelica language. Models for the different components were derived, suiting the purpose of this study. The equivalent parameters were derived from data sheets and other simulation programs such as PSCAD. The parameters were entered into the model grid and tested at steady state, firstly. This yielded satisfactory results that were similar to the reference results from MATPOWER power flow. Furthermore, fault conditions at several buses were simulated to observe the behaviour of the grid under these conditions. Recommendations for further developing this model to include more detailed models for components, such as power electronic converters, were made at the end of the thesis.

Simulation and Implementation of a PMSM Sensorless Control Algorithm for Submarine Applications

In this thesis, the study and the simulation of two advanced sensorless speed control techniques for a surface PMSM are presented. The aim is to implement a sensorless control algorithm for a submarine auxiliary propulsion system. This experimental activity is the result of a project collaboration with L3Harris Calzoni, a leader company in A&D systems for naval handling in military field. A Simulink model of the whole electric drive has been developed. Due to the satisfactory results of the simulations, the sensorless control system has been implemented in C code for STM32 environment. Finally, several tests on a real brushless machine have been carried out while the motor was connected to a mechanical load to simulate the real scenario of the final application. All the experimental results have been recorded through a graphical interface software developed at Calzoni.