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 visible light communications in vehicular networks

The rapid development in the field of lighting and illumination allows low energy consumption and a rapid growth in the use, and development of solid-state sources. As the efficiency of these devices increases and their cost decreases there are predictions that they will become the dominant source for general illumination in the short term. The objective of this thesis is to study, through extensive simulations in realistic scenarios, the feasibility and exploitation of visible light communication (VLC) for vehicular ad hoc networks (VANETs) applications. A brief introduction will introduce the new scenario of smart cities in which visible light communication will become a fundamental enabling technology for the future communication systems. Specifically, this thesis focus on the acquisition of several, frequent, and small data packets from vehicles, exploited as sensors of the environment. The use of vehicles as sensors is a new paradigm to enable an efficient environment monitoring and an improved traffic management. In most cases, the sensed information must be collected at a remote control centre and one of the most challenging aspects is the uplink acquisition of data from vehicles. My thesis discusses the opportunity to take advantage of short range vehicle-to-vehicle (V2V) and vehicle-to-roadside (V2R) communications to offload the cellular networks. More specifically, it discusses the system design and assesses the obtainable cellular resource saving, by considering the impact of the percentage of vehicles equipped with short range communication devices, of the number of deployed road side units, and of the adopted routing protocol. When short range communications are concerned, WAVE/IEEE 802.11p is considered as standard for VANETs. Its use together with VLC will be considered in urban vehicular scenarios to let vehicles communicate without involving the cellular network. The study is conducted by simulation, considering both a simulation platform (SHINE, simulation platform for heterogeneous interworking networks) developed within the Wireless communication Laboratory (Wilab) of the University of Bologna and CNR, and network simulator (NS3). trying to realistically represent all the wireless network communication aspects. Specifically, simulation of vehicular system was performed and introduced in ns-3, creating a new module for the simulator. This module will help to study VLC applications in VANETs. Final observations would enhance and encourage potential research in the area and optimize performance of VLC systems applications in the future.

Integration of ROS for the control of industrial robotic systems in a proprietary environment

The paper deals with the integration of ROS, in the proprietary environment of the Marchesini Group company, for the control of industrial robotic systems. The basic tools of this open-source software are deeply studied to model a full proprietary Pick and Place manipulator inside it, and to develop custom ROS nodes to calculate trajectories; speaking of which, the URDF format is the standard to represent robots in ROS and the motion planning framework MoveIt offers user-friendly high-level methods. The communication between ROS and the Marchesini control architecture is established using the OPC UA standard; the tasks computed are transmitted offline to the PLC, supervisor controller of the physical robot, because the performances of the protocol don’t allow any kind of active control by ROS. Once the data are completely stored at the Marchesini side, the industrial PC makes the real robot execute a trajectory computed by MoveIt, so that it replicates the behaviour of the simulated manipulator in Rviz. Multiple experiments are performed to evaluate in detail the potential of ROS in the planning of movements for the company proprietary robots. The project ends with a small study regarding the use of ROS as a simulation platform. First, it is necessary to understand how a robotic application of the company can be reproduced in the Gazebo real world simulator. Then, a ROS node extracts information and examines the simulated robot behaviour, through the subscription to specific topics.

Graph-based User Scheduling for MIMO LEO-based Satellite Communication Systems

The study of the user scheduling problem in a Low Earth Orbit (LEO) Multi-User MIMO system is the objective of this thesis. With the application of cutting-edge digital beamforming algorithms, a LEO satellite with an antenna array and a large number of antenna elements can provide service to many user terminals (UTs) in full frequency reuse (FFR) schemes. Since the number of UTs on-ground are many more than the transmit antennas on the satellite, user scheduling is necessary. Scheduling can be accomplished by grouping users into different clusters: users within the same cluster are multiplexed and served together via Space Division Multiple Access (SDMA), i.e., digital beamforming or Multi-User MIMO techniques; the different clusters of users are then served on different time slots via Time Division Multiple Access (TDMA). The design of an optimal user grouping strategy is known to be an NP-complete problem which can be solved only through exhaustive search. In this thesis, we provide a graph-based user scheduling and feed space beamforming architecture for the downlink with the aim of reducing user inter-beam interference. The main idea is based on clustering users whose pairwise great-circle distance is as large as possible. First, we create a graph where the users represent the vertices, whereas an edge in the graph between 2 users exists if their great-circle distance is above a certain threshold. In the second step, we develop a low complex greedy user clustering technique and we iteratively search for the maximum clique in the graph, i.e., the largest fully connected subgraph in the graph. Finally, by using the 3 aforementioned power normalization techniques, a Minimum Mean Square Error (MMSE) beamforming matrix is deployed on a cluster basis. The suggested scheduling system is compared with a position-based scheduler, which generates a beam lattice on the ground and randomly selects one user per beam to form a cluster.

Development of iodine and air chemistry models for the simulation of plasma in Helicon Plasma Thrusters

The aim of this work is to analyse the chemistry models of low pressure Helicon discharges fed with iodine and air. In particular the focus of this research is to understand the plasma dynamics in order to predict propulsive performances of iodine and air-breathing Helicon Plasma Thrusters. The two systems have been simulated and analysed with the use of global models, i.e. a 0 dimensional tool to solve the set of governing equations by assuming that all quantities are volume averaged. Furthermore, some strategies have been implemented to improve the accuracy of this approach. A verification have been accomplished on the global models for both iodine and air, comparing results against simulations taken from literature. Moreover, the iodine global model has been validated against the experimental measurements of REGULUS, an helicon plasma thruster developed by the Italian company T4i, with a good agreement. From the analysis of iodine model, it has been found a significantly higher density for atomic positive ions with respect to molecular ions. Negative ions, instead, have shown to have negligible effect on the propulsive results. Also, the influence of reactions between heavy particles has been analysed with the global model. Results have demonstrated that, in the iodine case, chemistry is almost entirely affected by electronic collisions. For what concerns air-breathing results, it has been investigated the effects of the orbital height on propulsive performances. In particular, the global model has shown that at lower height, the values of thrust and specific impulse are lower due a change in atmosphere concentration. Finally, the iodine chemistry model has been introduced in the fluid code 3D-VIRTUS in order to preliminary assess the plasma properties of a Helicon discharge chamber for electric propulsion.