Preliminary studies for the establishment of a tsunami early detection algorithm to be used in the frame of a Tsunami Warning System

This thesis presents and discusses TEDA, an algorithm for the automatic detection in real-time of tsunamis and large amplitude waves on sea level records. TEDA has been developed in the frame of the Tsunami Research Team of the University of Bologna for coastal tide gauges and it has been calibrated and tested for the tide gauge station of Adak Island, in Alaska. A preliminary study to apply TEDA to offshore buoys in the Pacific Ocean is also presented.

Non Invasive Tools for Early Detection of Autism Spectrum Disorders

Autism Spectrum Disorders (ASDs) describe a set of neurodevelopmental disorders. ASD represents a significant public health problem. Currently, ASDs are not diagnosed before the 2nd year of life but an early identification of ASDs would be crucial as interventions are much more effective than specific therapies starting in later childhood. To this aim, cheap an contact-less automatic approaches recently aroused great clinical interest. Among them, the cry and the movements of the newborn, both involving the central nervous system, are proposed as possible indicators of neurological disorders. This PhD work is a first step towards solving this challenging problem. An integrated system is presented enabling the recording of audio (crying) and video (movements) data of the newborn, their automatic analysis with innovative techniques for the extraction of clinically relevant parameters and their classification with data mining techniques. New robust algorithms were developed for the selection of the voiced parts of the cry signal, the estimation of acoustic parameters based on the wavelet transform and the analysis of the infant’s general movements (GMs) through a new body model for segmentation and 2D reconstruction. In addition to a thorough literature review this thesis presents the state of the art on these topics that shows that no studies exist concerning normative ranges for newborn infant cry in the first 6 months of life nor the correlation between cry and movements. Through the new automatic methods a population of control infants (“low-risk”, LR) was compared to a group of “high-risk” (HR) infants, i.e. siblings of children already diagnosed with ASD. A subset of LR infants clinically diagnosed as newborns with Typical Development (TD) and one affected by ASD were compared. The results show that the selected acoustic parameters allow good differentiation between the two groups. This result provides new perspectives both diagnostic and therapeutic.

Relevance of the pig stomach for the detection of dietary factors and gut maturation and control

In this thesis two approaches were applied to achieve a double general objective. The first chapter was dedicated to the study of the distribution of the expression of genes of several bitter and fat receptor in several gastrointestinal tracts. A set of 7 genes for bitter taste and for 3 genes for fat taste was amplified with real-time PCR from mRNA extracted from 5 gastrointestinal segments of weaned pigs. The presence of gene expression for several chemosensing receptors for bitter and fat taste in different compartments of the stomach confirms that this organ should be considered a player for the early detection of bolus composition. In the second chapter we investigated in young pigs the distribution of butyrate-sensing olfactory receptor (OR51E1) receptor along the GIT, its relation with some endocrine markers, its variation with age, and after interventions affecting the gut environment and intestinal microbiota in piglets and in different tissues. Our results indicate that OR51E1 is strictly related to the normal GIT enteroendocrine activity. In the third chapter we investigated the differential gene expression between oxyntic and pyloric mucosa in seven starter pigs. The obtained data indicate that there is significant differential gene exression between oxintic of the young pig and pyloric mucosa and further functional studies are needed to confirm their physiological importance. In the last chapter, thymol, that has been proposed as an oral alternative to antibiotics in the feed of pigs and broilers, was introduced directly into the stomach of 8 weaned pigs and sampled for gastric oxyntic and pyloric mucosa. The analysis of the whole transcript expression shoes that the stimulation of gastric proliferative activity and the control of digestive activity by thymol can influence positively gastric maturation and function in the weaned pigs.

Sviluppo di un tomografo multi-energy per lo studio pre-clinico di nuove metodiche diagnostiche finalizzate al riconoscimento precoce della patologia tumorale

A new multi-energy CT for small animals is being developed at the Physics Department of the University of Bologna, Italy. The system makes use of a set of quasi-monochromatic X-ray beams, with energy tunable in a range from 26 KeV to 72 KeV. These beams are produced by Bragg diffraction on a Highly Oriented Pyrolytic Graphite crystal. With quasi-monochromatic sources it is possible to perform multi-energy investigation in a more effective way, as compared with conventional X-ray tubes. Multi-energy techniques allow extracting physical information from the materials, such as effective atomic number, mass-thickness, density, that can be used to distinguish and quantitatively characterize the irradiated tissues. The aim of the system is the investigation and the development of new pre-clinic methods for the early detection of the tumors in small animals. An innovative technique, the Triple-Energy Radiography with Contrast Medium (TER), has been successfully implemented on our system. TER consist in combining a set of three quasi-monochromatic images of an object, in order to obtain a corresponding set of three single-tissue images, which are the mass-thickness map of three reference materials. TER can be applied to the quantitative mass-thickness-map reconstruction of a contrast medium, because it is able to remove completely the signal due to other tissues (i.e. the structural background noise). The technique is very sensitive to the contrast medium and is insensitive to the superposition of different materials. The method is a good candidate to the early detection of the tumor angiogenesis in mice. In this work we describe the tomographic system, with a particular focus on the quasi-monochromatic source. Moreover the TER method is presented with some preliminary results about small animal imaging.

Sorveglianza dell'influenza aviare: studio di un sistema di rilevazione precoce della circolazione virale in popolazioni di volatili selvatici

The emergency of infection by highly pathogenic avian influenza virus (HPAI) subtype H5N1 has focused the attention of the world scientific community, requiring the prompt provision of effective control systems for early detection of the circulation of low pathogenic influenza H5 viruses (LPAI) in populations of wild birds to prevent outbreaks of highly pathogenic (HPAI) in populations of domestic birds with possible transmission to humans. The project stems from the aim to provide, through a preliminary analysis of data obtained from surveillance in Italy and Europe, a preliminary study about the virus detection rates and the development of mathematical models, an objective assessment of the effectiveness of avian influenza surveillance systems in wild bird populations, and to point out guidelines to support the planning process of the sampling activities. The results obtained from the statistical processing quantify the sampling effort in terms of time and sample size required, and simulating different epidemiological scenarios identify active surveillance as the most suitable for endemic LPAI infection monitoring in wild waterfowl, and passive surveillance as the only really effective tool in early detecting HPAI H5N1 circulation in wild populations. Given the lack of relevant information on H5N1 epidemiology, and the actual finantial and logistic constraints, an approach that makes use of statistical tools to evaluate and predict monitoring activities effectiveness proves to be of primary importance to direct decision-making and make the best use of available resources.

New approach in the diagnosis and therapy of hyperphenylalaninemia

Background. Phenylketonuria is the most prevalent inborn error of aminoacid metabolism. Is an autosomal recessive disorder. It results from mutations in the phenylalanine hydroxilase (PAH) gene. Phenotypes can vary from mild hyperphenylalaninemia to a severe phenylketonuria wich, if untreated, results in severe mental retardation. Thanks to neonatal screening programmes, early detection and promp dietetic intervention (phenylalanine restricted diet lifelong) has allowed to avoid neurocognitive complications. Recently, a new therapy is become widely used: the oral supplementation with the PAH cofactor (BH4), wich can alleviate the diet burden. Genotype-phenotype correlation is a reliable tool to predict metabolic phenotype in order to establish a better tailored diet and to assess the potential responsiveness to BH4 therapy. Aim Molecular analysis of the PAH gene, evaluation of genotype-phenotype correlation and prediction of BH4 responsiveness in a group of HPA patients living in Emilia Romagna. Patients and methods. We studied 48 patients affected by PAH deficiency in regular follow-up to our Metabolic Centre. We performed the molecular analysis of these patients using genomic DNA extracted from peripheral blood samples Results. We obtained a full genotipic characterization of 46 patients. We found 87 mutant alleles and 35 different mutations, being the most frequent IVS10-11 G>A (19.3%), R261Q (9.1%), R158Q (9.1%), R408Q (6.8%) and A403V (5.7%), including 2 new ones (L287, N223Y) ever described previously. Notably, we found 15 mutations already identified in BH4-responsive patients, according to the literature. We found 42 different genotipic combinations, most of them in single patients and involving a BH4-responsive mutation. Conclusion. BH4 responsiveness is shown by a consistent number of PAH deficient hyperphenylalaninemic patients. This treatment, combined with a less restricted diet or as monotherapy, can reduce nutritional complications and improve the quality of life of these patients.

Nanotechnology control of self organized biomolecules and biomaterials for medical research

Nanotechnology entails the manufacturing and manipulation of matter at length scales ranging from single atoms to micron-sized objects. The ability to address properties on the biologically-relevant nanometer scale has made nanotechnology attractive for Nanomedicine. This is perceived as a great opportunity in healthcare especially in diagnostics, therapeutics and more in general to develop personalized medicine. Nanomedicine has the potential to enable early detection and prevention, and to improve diagnosis, mass screening, treatment and follow-up of many diseases. From the biological standpoint, nanomaterials match the typical size of naturally occurring functional units or components of living organisms and, for this reason, enable more effective interaction with biological systems. Nanomaterials have the potential to influence the functionality and cell fate in the regeneration of organs and tissues. To this aim, nanotechnology provides an arsenal of techniques for intervening, fabricate, and modulate the environment where cells live and function. Unconventional micro- and nano-fabrication techniques allow patterning biomolecules and biocompatible materials down to the level of a few nanometer feature size. Patterning is not simply a deterministic placement of a material; in a more extended acception it allows a controlled fabrication of structures and gradients of different nature. Gradients are emerging as one of the key factors guiding cell adhesion, proliferation, migration and even differentiation in the case of stem cells. The main goal of this thesis has been to devise a nanotechnology-based strategy and tools to spatially and temporally control biologically-relevant phenomena in-vitro which are important in some fields of medical research.

Biomimetic Scaffolds for the Controlled Release of Bioactive Molecules for Tissue Engineering Applications

The temporospatial controlled delivery of growth factors is crucial to trigger the desired healing mechanisms in target tissues. The uncontrolled release of growth factors has been demonstrated to cause severe side effects in its surrounding tissues. Thus, the first working hypothesis was to tune and optimize a newly developed multiscale delivery platform based on a nanostructured silicon particle core (pSi) and a poly (dl-lactide-co-glycolide) acid (PLGA) outer shell. In a murine subcutaneous model, the platform was demonstrated to be fully tunable for the temporal and spatial control release of the payload. Secondly, a multiscale approach was followed in a multicompartment collagen scaffold, to selectively integrate different sets of PLGA-pSi loaded with different reporter proteins. The spatial confinement of the microspheres allowed the release of the reporter proteins in each of the layers of the scaffold. Finally, the staged and zero-order release kinetics enabled the temporal biochemical patterning of the scaffold. The last step of this PhD project was to test if by fully embedding PLGA microspheres in a highly structured and fibrous collagen-based scaffold (camouflaging), it was possible to prevent their early detection and clearance by macrophages. It was further studied whether such a camouflaging strategy was efficient in reducing the production of key inflammatory molecules, while preserving the release kinetics of the payload of the PLGA microspheres. Results demonstrated that the camouflaging allowed for a 10-fold decrease in the number of PLGA microspheres internalized by macrophages, suggesting that the 3D scaffold operated by cloaking the PLGA microspheres. When the production of key inflammatory cytokines induced by the scaffold was assessed, macrophages' response to the PLGA microspheres-integrated scaffolds resulted in a response similar to that observed in the control (not functionalized scaffold) and the release kinetic of a reporter protein was preserved.

Sviluppo di mezzi e sistemi di sorveglianza e monitoraggio dei ditteri culicidi di importanza sanitaria

Nel corso degli ultimi anni le problematiche legate al ruolo vettore delle zanzare stanno emergendo sia per quanto riguarda l’uomo che gli animali allevati e selvatici. Diversi arbovirus come West Nile, Chikungunya, Usutu e Dengue, possono facilmente spostarsi a livello planetario ed essere introdotti anche nei nostri territori dove possono dare avvio a episodi epidemici. Le tecniche di monitoraggio e sorveglianza dei Culicidi possono essere convenientemente utilizzate per il rilevamento precoce dell’attività virale sul territorio e per la stima del rischio di epidemie al fine dell’adozione delle opportune azioni di Sanità Pubblica. Io scopo della ricerca del dottorato è inserito nel contesto dei temi di sviluppo del Piano regionale sorveglianza delle malattie trasmesse da vettori in Emilia Romagna. La ricerca condotta è inquadrata prevalentemente sotto l’aspetto entomologico applicativo di utilizzo di dispositivi (trappole) che possano catturare efficacemente possibili insetti vettori. In particolare questa ricerca è stata mirata allo studio comparativo in campo di diversi tipi di trappole per la cattura di adulti di zanzara, cercando di interpretare i dati per capire un potenziale valore di efficacia/efficienza nel rilevamento della circolazione virale e come supporto alla pianificazione della rete di sorveglianza dal punto di vista operativo mediante dispositivi adeguati alle finalità d’indagine. Si è cercato di trovare un dispositivo idoneo, approfondendone gli aspetti operativi/funzionali, ai fini di cattura del vettore principale del West Nile Virus, cioè la zanzara comune, da affiancare all’unica tipologia di trappola usata in precedenza. Le prove saranno svolte sia in campo che presso il laboratorio di Entomologia Medica Veterinaria del Centro Agricoltura Ambiente “G. Nicoli” di Crevalcore, in collaborazione con il Dipartimento di Scienze e Tecnologie Agroambientali della Facoltà di Agraria dell’Università di Bologna.

Studi epidemiologici su Pseudomonas syringae pv.actinidiae agente causale del cancro batterico in actinidia

Il cancro batterico dell’actinidia causato da Pseudomonas syringae pv.actinidiae (Psa) suscita grande interesse a livello globale a partire dal 2008. La malattia è comparsa in Giappone e in due anni ha avuto una diffusione epidemica in tutte le aree di coltivazione mondiale di actinidia. Gravi perdite economiche hanno attirato l’attenzione internazionale su questa problematica e grandi sforzi sono stati rivolti allo studio di questo patosistema ancora poco conosciuto. E’ emerso infatti che il patogeno può rimanere in fase latente per lunghi periodi senza causare sintomi caratteristici nelle piante infette, e che dalla comparsa dei sintomi la pianta muore nell’arco di un paio d’anni. Il monitoraggio ed il controllo della situazione è perciò di fondamentale importanza ed è ancora più importante prevenire la comparsa di nuovi focolai di infezione. A questo proposito sarebbe opportuno l’impiego di materiale vegetale di propagazione non infetto, ma in molti casi questo diventa difficile, dal momento che il materiale impiegato è generalmente quello asintomatico, non analizzato precedentemente per la presenza del patogeno. Negli ultimi anni sono state perciò messe a punto molte tecniche molecolari per l’identificazione di Psa direttamente da materiale vegetale. L’obiettivo di questo lavoro è stato quello di studiare l’epidemiologia di Psa in piante adulte infette e di verificare l’efficacia di metodi di diagnosi precoce per prevenire la malattia. A tale scopo il lavoro sperimentale è stato suddiviso in diverse fasi: i) studio della localizzazione, traslocazione e sopravvivenza di Psa nelle piante, a seguito di inoculazione in piante adulte di actinidia di ceppi marcati Psa::gfp; ii) studio della capacità di Psa di essere mantenuto in germogli di actinidia attraverso sette generazioni di micropropagazione dopo l’inoculazione delle piante madri con lo stesso ceppo marcato Psa::gfp; iii) studio ed applicazioni di un nuovo metodo di diagnosi precoce di Psa basato sull’analisi molecolare del “pianto”.

Managing invasive populations of Anoplophora chinensis and A. glabripennis in Lombardy

Two Asian longhorned beetles (Coleoptera: Cerambycidae), commonly known as Citrus Longhorned Beetle (CLB), Anoplophora chinensis (Forster), and Asian Longhorned Beetle (ALB), A. glabripennis (Motschulsky), are considered the most destructive wood borers introduced in Lombardy (northern Italy). This research aimed at (1) improving laboratory rearing methods for the biological control agent Aprostocetus anoplophorae (Hym.: Eulophidae), an egg parasitoid specific to CLB, and defining release techniques allowing its establishment; (2) test the efficacy of the sentinel tree technique for the early detection of CLB; and (3) evaluating the efficacy of traps baited with artificial lures in attracting adults of ALB and possibly CLB. Several problems were faced while rearing the egg parasitoid in laboratory. It appeared that the rate of parasitism of the hosts could depend on the age of the host eggs and/or age of the laying parasitoid females. Data results from the field experiments about A. anoplophorae release-capture showed that the percentage of slits containing a CLB egg was particularly low on most sentinel trees and the percentage of CLB eggs that were killed, because of natural predators, was high. Only one egg amongst those exposed was attacked by the released parasitoid. These negative results were anyway very useful, since they provided evidence and information on the type of host plants to be used, the time necessary for the exposure of the plants to the egg-laying CLB females, the number of laying parasitoid females to be inserted per cage. The sentinel trees technique revealed to be not successful; signs and symptoms of CLB presence were not recorded during the two seasons of field observations (2012-2013). Extremely positive was instead the trial with artificial lures carried out during summer 2013. A total of 32 beetles were captured (4 ALB and 28 CLB) deploying 50 baited traps.

Fault detection in rotating machines by vibration signal processing techniques

Machines with moving parts give rise to vibrations and consequently noise. The setting up and the status of each machine yield to a peculiar vibration signature. Therefore, a change in the vibration signature, due to a change in the machine state, can be used to detect incipient defects before they become critical. This is the goal of condition monitoring, in which the informations obtained from a machine signature are used in order to detect faults at an early stage. There are a large number of signal processing techniques that can be used in order to extract interesting information from a measured vibration signal. This study seeks to detect rotating machine defects using a range of techniques including synchronous time averaging, Hilbert transform-based demodulation, continuous wavelet transform, Wigner-Ville distribution and spectral correlation density function. The detection and the diagnostic capability of these techniques are discussed and compared on the basis of experimental results concerning gear tooth faults, i.e. fatigue crack at the tooth root and tooth spalls of different sizes, as well as assembly faults in diesel engine. Moreover, the sensitivity to fault severity is assessed by the application of these signal processing techniques to gear tooth faults of different sizes.