Steel and macro-synthetic self-compacting fibre reinforced concrete, experimental study on the long-term deformations

Experimental study on the long-term deformations of the fibre reinforced concrete. Steel and macro-synthetic fibers were used to evaluate the shrinkage, creep, mid-span deflection, cracking and rupture analysis of three different types of samples. At the end the main topics of ACI guidelines were analyzed in order to perform an overview of design.

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.

Surface-enhanced vibrational techniques for the detection of synthetic organic colorants

The research project object of this thesis is focused on the development of an advanced analytical system based on the combination of an improved thin layer chromatography (TLC) plate coupled with infrared (FTIR) and Raman microscopies for the detection of synthetic dyes. Indeed, the characterization of organic colorants, which are commonly present in mixtures with other components and in a very limited amount, still represents a challenging task in scientific analyses of cultural heritage materials. The approach provides selective spectral fingerprints for each compound, foreseeing the complementary information obtained by micro ATR-RAIRS-FTIR and SERS-Raman analyses, which can be performed on the same separated spot. In particular, silver iodide (AgI) applied on a gold coated slide is proposed as an efficient stationary phase for the discrimination of complex analyte mixtures, such as dyes present in samples of art-historical interest. The gold-AgI-TLC plate shows high performances related both to the chromatographic separation of analytes and to the spectroscopic detection of components. The use of a mid-IR transparent inorganic salt as the stationary phase avoids interferences of the background absorption in FTIR investigations. Moreover, by ATR microscopy measurements performed on the gold-AgI surface, a considerable enhancement in the intensity of spectra is observed. Complementary information can be obtained by Raman analyses, foreseeing a SERS activity of the AgI substrate. The method has been tested for the characterization of a mixture of three synthetic organic colorants widely used in dyeing processes: Brilliant Green (BG1), Rhodamine B (BV10) and Methylene Blue (BB9).

Synthetic studies towards a new fulvestrant analogue

A study towards the synthesis of a new fulvestrant analogue with improved bioavailability was carried out. In this work a twelve-step synthetic route starting from β-estradiol was optimized and a palladium (Pd)-catalyzed endo-selective Heck reaction for the functionalization of an advanced intermediate was investigated.

Production Lot Detection through Synthetic Generated Data

Correctness of information gathered in production environments is an essential part of quality assurance processes in many industries, this task is often performed by human resources who visually take annotations in various steps of the production flow. Depending on the performed task the correlation between where exactly the information is gathered and what it represents is more than often lost in the process. The lack of labeled data places a great boundary on the application of deep neural networks aimed at object detection tasks, moreover supervised training of deep models requires a great amount of data to be available. Reaching an adequate large collection of labeled images through classic techniques of data annotations is an exhausting and costly task to perform, not always suitable for every scenario. A possible solution is to generate synthetic data that replicates the real one and use it to fine-tune a deep neural network trained on one or more source domains to a different target domain. The purpose of this thesis is to show a real case scenario where the provided data were both in great scarcity and missing the required annotations. Sequentially a possible approach is presented where synthetic data has been generated to address those issues while standing as a training base of deep neural networks for object detection, capable of working on images taken in production-like environments. Lastly, it compares performance on different types of synthetic data and convolutional neural networks used as backbones for the model.

Synthetic data generation for the assessment of antimicrobial resistance through machine learning

As a consequence of the diffusion of next generation sequencing techniques, metagenomics databases have become one of the most promising repositories of information about features and behavior of microorganisms. One of the subjects that can be studied from those data are bacteria populations. Next generation sequencing techniques allow to study the bacteria population within an environment by sampling genetic material directly from it, without the needing of culturing a similar population in vitro and observing its behavior. As a drawback, it is quite complex to extract information from those data and usually there is more than one way to do that; AMR is no exception. In this study we will discuss how the quantified AMR, which regards the genotype of the bacteria, can be related to the bacteria phenotype and its actual level of resistance against the specific substance. In order to have a quantitative information about bacteria genotype, we will evaluate the resistome from the read libraries, aligning them against CARD database. With those data, we will test various machine learning algorithms for predicting the bacteria phenotype. The samples that we exploit should resemble those that could be obtained from a natural context, but are actually produced by a read libraries simulation tool. In this way we are able to design the populations with bacteria of known genotype, so that we can relay on a secure ground truth for training and testing our algorithms.

Bitumen aggregate adhesion and bitumen rheology of hard modified binders

The waterproofing of hydraulic structure is done traiditionally like laying road on dam surface but with specific modified binders. An italian firm recently patented a new method that is re-adaptation of typical surface treatment of roads. The purpose of this study is to find out best aggregate-bitumen mixture that can perform well under service conditions of a large hydraulic structure such as dams. So, 4 different hard modified bitumen were tested with 2 aggregate types i.e. limestone and basalt. The experimental program contained the testing of bitumen aggregate adhesion, using the rolling bottle test and rheology of hard modified binders using multiple stress creep and recovery test and dynamic viscosity test. The results and discussion are presented in detail in this work.

Synthetic generation of amyloid PET images by non-linear dimensionality reduction inversion

Privacy issues and data scarcity in PET field call for efficient methods to expand datasets via synthetic generation of new data that cannot be traced back to real patients and that are also realistic. In this thesis, machine learning techniques were applied to 1001 amyloid-beta PET images, which had undergone a diagnosis of Alzheimer’s disease: the evaluations were 540 positive, 457 negative and 4 unknown. Isomap algorithm was used as a manifold learning method to reduce the dimensions of the PET dataset; a numerical scale-free interpolation method was applied to invert the dimensionality reduction map. The interpolant was tested on the PET images via LOOCV, where the removed images were compared with the reconstructed ones with the mean SSIM index (MSSIM = 0.76 ± 0.06). The effectiveness of this measure is questioned, since it indicated slightly higher performance for a method of comparison using PCA (MSSIM = 0.79 ± 0.06), which gave clearly poor quality reconstructed images with respect to those recovered by the numerical inverse mapping. Ten synthetic PET images were generated and, after having been mixed with ten originals, were sent to a team of clinicians for the visual assessment of their realism; no significant agreements were found either between clinicians and the true image labels or among the clinicians, meaning that original and synthetic images were indistinguishable. The future perspective of this thesis points to the improvement of the amyloid-beta PET research field by increasing available data, overcoming the constraints of data acquisition and privacy issues. Potential improvements can be achieved via refinements of the manifold learning and the inverse mapping stages during the PET image analysis, by exploring different combinations in the choice of algorithm parameters and by applying other non-linear dimensionality reduction algorithms. A final prospect of this work is the search for new methods to assess image reconstruction quality.