Teses / dissertações sobre o tema "303.48/3/08"

“Il Presidente del Consiglio dei Ministri nell'ordinamento vigente e nelle proposte di riforma”. La tesi si propone di ricostruire i caratteri propri della figura giuridica del Presidente del Consiglio nell’ordinamento vigente, sino alla analisi delle più recenti proposte di riforma costituzionale in tema. La dimensione analitico-sistematica proposta è stata sviluppata in riferimento alle concrete dinamiche economiche, politiche ed istituzionali, in relazione alle quali si è inteso verificare il tipo di rapporto tra esigenze di stabilizzazione del ciclo economico ed esigenze di stabilizzazione degli esecutivi, dando il dovuto rilievo al processo di integrazione della Unione Europea, come “motore” delle istanze di riforma del sistema economico ed istituzionale. Nel primo capitolo, si sono esplicitate le chiavi metodologiche secondo le quali si è inteso sviluppare il percorso di ricerca. Nel secondo capitolo si è operata una ricostruzione del quadro ordinamentale a base statutaria, come termine iniziale per poter cogliere al meglio la configurazione del ruolo poi assunta nell’ordinamento repubblicano; ciò come premessa necessaria ad una corretta ricostruzione diacronica, di tutti gli elementi rilevanti per poter comprendere la portata delle proposte attuali. Nel terzo capitolo, si è cercato di ricostruire il ruolo del presidente del consiglio, collocandolo nel quadro della dinamica attuazione/inattuazione dell’ordinamento repubblicano e alla luce dei caratteri del sistema politico, in modo da poter coniugare i tratti evolutivi imposti all’inquadramento della figura dalle dinamiche del sistema politico; così da poter conseguentemente cogliere gli elementi genetici di un dibattito politico istituzionale, che genera proposte di riforma organica, mediante l’analisi degli atti delle commissioni Parlamentari per le riforme. Nel quarto capitolo, a partire dalla apertura di una fase di c.d. “transizione”, si ricostruiscono le proposte specificamente finalizzate al riassetto del Presidente del Consiglio prodotte dalle commissioni parlamentari per le riforme, segnando le tappe di un percorso ancora aperto e dall’esito incerto.
The work analyses the institutional characters of the head of the government, in particular appointing the powers about his ministers and the powers of the government in Parliament, in connections with the legislative procedures. The analysis shows the importance of the party-system in conditioning the balance bitwin parliament and government. In particular, a plularistic asset of the party-sistem, based on a proportional electoral law, favourishes the weekness of the government. On this considerations, the istances of reform are oriented to enforce the government and simplify the party-system, in order to give as a result a “deciding democracy”, based on a strength Parliament and a strength Government.

Nel 2018, alcuni professionisti dell’Azienda Sant’Orsola-Malpighi e dell’Università di Bologna hanno costituito un gruppo multidisciplinare con lo scopo di fornire una diagnosi eziologica nei casi di morte cardiaca improvvisa (MCI). Tutti i casi di MCI in soggetti di età > 1 anno e ≤ 55 anni sono stati sottoposti ad uno studio anatomo-patologico del cuore presso l’Unità di Patologia Cardiovascolare del Sant’Orsola-Malpighi. I casi con sospetta malattia genetica e i casi con cuore strutturalmente normale sono stati avviati ad analisi presso la Genetica Medica. In alcuni casi è stato necessario procedere ad indagini tossicologiche o microbiologiche. Ai familiari dei pazienti deceduti è stato successivamente offerto uno screening cardiologico clinico e strumentale. Nei due anni di attività sono giunti all’attenzione dell’Unità di Patologia Cardiovascolare 50 casi di MCI. L’età media dei soggetti è risultata 35±13.6 anni. Il 100% dei casi è stato sottoposto ad esame cardio-patologico completo, il 64% dei casi è stato sottoposto ad analisi genetica, il 44% ad indagini tossicologiche e il 4% ad indagini microbiologiche. L’approccio scelto ha permesso di raggiungere un elevato grado di certezza diagnostica nell’85.5% dei casi e, in particolare, una diagnosi certa è stata posta nel 60.5% dei casi. Nella nostra casistica le malattie non ischemiche del miocardio rappresentano la causa più frequente di MCI (39.9%). Fra queste le miocarditi/malattie infiammatorie del miocardio, sono il 42.3%. Le malattie coronariche sono risultate la causa della MCI nel 23.1% dei casi e la patologia dissecativa delle coronarie costituiva una percentuale consistente di questo sottogruppo (20%). I cuori strutturalmente normali rappresentano il 14.5% della nostra casistica. Le indagini tossicologiche hanno fornito un importante contributo. In 10 famiglie sono state proposte indagini di screening ai parenti di primo grado.
Nel 2018, alcuni professionisti dell’Azienda Sant’Orsola-Malpighi e dell’Università di Bologna hanno costituito un gruppo multidisciplinare con lo scopo di fornire una diagnosi eziologica nei casi di morte cardiaca improvvisa (MCI). Tutti i casi di MCI in soggetti di età > 1 anno e ≤ 55 anni sono stati sottoposti ad uno studio anatomo-patologico del cuore presso l’Unità di Patologia Cardiovascolare del Sant’Orsola-Malpighi. I casi con sospetta malattia genetica e i casi con cuore strutturalmente normale sono stati avviati ad analisi presso la Genetica Medica. In alcuni casi è stato necessario procedere ad indagini tossicologiche o microbiologiche. Ai familiari dei pazienti deceduti è stato successivamente offerto uno screening cardiologico clinico e strumentale. Nei due anni di attività sono giunti all’attenzione dell’Unità di Patologia Cardiovascolare 50 casi di MCI. L’età media dei soggetti è risultata 35±13.6 anni. Il 100% dei casi è stato sottoposto ad esame cardio-patologico completo, il 64% dei casi è stato sottoposto ad analisi genetica, il 44% ad indagini tossicologiche e il 4% ad indagini microbiologiche. L’approccio scelto ha permesso di raggiungere un elevato grado di certezza diagnostica nell’85.5% dei casi e, in particolare, una diagnosi certa è stata posta nel 60.5% dei casi. Nella nostra casistica le malattie non ischemiche del miocardio rappresentano la causa più frequente di MCI (39.9%). Fra queste le miocarditi/malattie infiammatorie del miocardio, sono il 42.3%. Le malattie coronariche sono risultate la causa della MCI nel 23.1% dei casi e la patologia dissecativa delle coronarie costituiva una percentuale consistente di questo sottogruppo (20%). I cuori strutturalmente normali rappresentano il 14.5% della nostra casistica. Le indagini tossicologiche hanno fornito un importante contributo. In 10 famiglie sono state proposte indagini di screening ai parenti di primo grado.

In this thesis, we are interested in approximating, by model order reduction, the solution to large-scale matrix- or tensor-valued semilinear Ordinary Differential Equations (ODEs). Under specific hypotheses on the linear operators and the considered domain, these ODEs often stem from the space discretization on a tensor basis of semilinear Partial Differential Equations (PDEs) with a dimension greater than or equal to two. The bulk of this thesis is devoted to the case where the discrete system is a matrix equation. We consider separately the cases of general Lipschitz continuous nonlinear functions and the Differential Riccati Equation (DRE) with a quadratic nonlinear term. In both settings, we construct a pair of left-right approximation spaces that leads to a reduced semilinear matrix differential equation with the same structure as the original problem, which can be more rapidly integrated with matrix-oriented integrators. For the DRE, under certain assumptions on the data, we show that a reduction process onto rational Krylov subspaces obtains significant computational and memory savings as opposed to current approaches. In the more general setting, a challenging difference lies in selecting and constructing the two approximation bases to handle the nonlinear term effectively. In addition, the nonlinear term also needs to be approximated for efficiency. To this end, in the framework of the Proper Orthogonal Decomposition (POD) methodology and the Discrete Empirical Interpolation Method (DEIM), we derive a novel matrix-oriented reduction process leading to a practical, structure-aware low order approximation of the original problem. In the final part of the thesis, we consider the multidimensional setting. Here we extend the matrix-oriented POD-DEIM algorithm to the tensor setting and illustrate how we can apply it to systems of such equations. Moreover, we discuss how to integrate the reduced-order model and, in particular, how to solve the resulting tensor-valued linear systems.

In these last years a great effort has been put in the development of new techniques for automatic object classification, also due to the consequences in many applications such as medical imaging or driverless cars. To this end, several mathematical models have been developed from logistic regression to neural networks. A crucial aspect of these so called classification algorithms is the use of algebraic tools to represent and approximate the input data. In this thesis, we examine two different models for image classification based on a particular tensor decomposition named Tensor-Train (TT) decomposition. The use of tensor approaches preserves the multidimensional structure of the data and the neighboring relations among pixels. Furthermore the Tensor-Train, differently from other tensor decompositions, does not suffer from the curse of dimensionality making it an extremely powerful strategy when dealing with high-dimensional data. It also allows data compression when combined with truncation strategies that reduce memory requirements without spoiling classification performance. The first model we propose is based on a direct decomposition of the database by means of the TT decomposition to find basis vectors used to classify a new object. The second model is a tensor dictionary learning model, based on the TT decomposition where the terms of the decomposition are estimated using a proximal alternating linearized minimization algorithm with a spectral stepsize.
Negli ultimi anni si è registrato un notevole sviluppo di nuove tecniche per il riconoscimento automatico di oggetti, anche dovuto alle possibili ricadute di tali avanzamenti nel campo medico o automobilistico. A tal fine sono stati sviluppati svariati modelli matematici dai metodi di regressione fino alle reti neurali. Un aspetto cruciale di questi cosiddetti algoritmi di classificazione è l'uso di aspetti algebrici per la rappresentazione e l'approssimazione dei dati in input. In questa tesi esamineremo due diversi modelli per la classificazione di immagini basati sulla decomposizione Tensor-Train (TT). In generale, l'uso di approcci tensoriali è fondamentale per preservare la struttura intrinsecamente multidimensionale dei dati. Inoltre l'occupazione di memoria per la decomposizione Tensor-Train non cresce esponenzialmente all'aumentare dei dati, a differenza di altre decomposizioni tensoriali. Questo la rende particolarmente adatta nel caso di dati di grandi dimensioni. Inoltre permette, attraverso l'uso di opportune strategie di troncamento, di limitare notevolmente l'occupazione di memoria senza ricadute negative sulle performance di classificazione. Il primo modello proposto in questa tesi è basato su una decomposizione diretta del database tramite la decomposizione TT. In questo modo viene determinata una base che verrà di seguito utilizzata nella classificazione di nuove immagini. Il secondo è invece un modello di dictionary learning tensoriale sempre basato sulla decomposizione TT in cui i termini della decomposizione sono determinati utilizzando un nuovo metodo di ottimizzazione alternato con l'utilizzo di passi spettrali.

In heroin fatalities the diagnosis of the cause of death may be particularly difficult because of several reasons, such as the relationship between lethal dose and current individual tolerance, the complexity of heroin metabolism, the presence of systemic dysfunction, and the contemporary use of other drugs of abuse. Thus, a wide variability is present in post-mortem blood concentration of morphine (MOR), the main metabolite of heroin, which is usually the most important analytical result for the interpretation of the cause of death. Recently, increasing interest has grown towards the role of the metabolites morphine-3-β-D-glucuronide (M3G) and morphine-6-β-D-glucuronide (M6G) in mediating heroin effects. The aim of this PhD study has been the development of a LC/MS-MS method for the determination of MOR, M3G and M6G in autopsy blood samples. An ESI-QqQ Mass Spectrometer, operating in positive ionisation and MRM mode, was used. Chromatographic separation was achieved thanks to a Reversed-Phase method, using a C18 column and a gradient elution with a binary mobile phase. SPE technique was employed to extract the analytes from biological samples. After validation, the method was applied to twenty-five blood specimens collected from cases of suspected fatal heroin overdose. The concentrations and the molar ratios of MOR, M3G and M6G were investigated. The influence of some risk factors, such as the contemporary use of alcohol, methadone or cocaine, was also studied.

Introduction: Behavioral lifestyle interventions focused on diet and physical activity are a cornerstone for the treatment of obesity. However, their effects vary substantially across individuals in terms of magnitude and durability. Personalized approaches that target psychological well-being may facilitate healthy behaviors and sustained weight loss. Objectives: This study aimed to explore whether the sequential combination of behavioral lifestyle and well-being intervention (BLI/WBI) may result in more favorable outcomes than behavioral lifestyle intervention alone (BLIA) in promoting weight loss (primary outcome) and improving psychological well-being, distress, dietary behaviors, and physical activity (secondary outcomes). Methods: 83 patients with obesity were randomly assigned to the BLI/WBI group (n=38) or the BLIA group (n=43). Participants in the BLIA group received a 12-week behavioral weight loss program, while those in the BLI/WBI received the same program followed by an additional 4-week well-being intervention. Data were collected at pretreatment (baseline, T1), at the end of BLI/WBI (T2), and at 6-month follow-up (T3). Results: There was a significant weight loss in both treatment groups at the end of BLI/WBI and 6-month follow-up. The BLI/WBI group showed greater improvements in anxiety symptoms (β=-1.92 [-3.66 to -0.18], p< 0.05) at the end of BLI/WBI and in depressive symptoms (β= -2.28[-4.20 to -0.37], p< 0.05) at 6-month follow-up compared to the BLIA group. Conclusions: The WBI showed no additional effect on weight loss. However, the secondary outcomes indicate that the WBI may reduce vulnerability to depressive and anxiety symptoms in obese patients. Future studies are needed to explore whether the psychological benefits of WBI can foster long-term weight loss maintenance.

Tumor heterogeneity and complex resistance mechanisms undermine the therapeutic value of existing anticancer options, generating a pressing demand for alternative accessible and effective leads. In this sense, our group recently introduced 3-chloropiperidines (3-CePs) as a novel class of alkylating agents designed to improve the pharmacological profile of nitrogen mustard (NM) chemotherapeutics. Previous works described the improved alkylating properties of bifunctional 3-chloropiperidines (B-CePs) compared to the reference NM chlorambucil, exploring the effect of different linker structures on their reactivity with DNA and dissecting the mechanism of B-CePs alkylation at guanines. From these premises, this project further investigated the chemical space and antiproliferative value of these anticancer candidates. First, an explorative set of monofunctional analogues (M-CePs) was introduced to study the effect of a single reactive center on the activity of compounds. Notably, despite reduced DNA cleavage relative to the bifunctional compounds, these new agents showed nanomolar cytotoxicity indexes against test cancer cells, along with an unexpected tropism for a pancreatic adenocarcinoma cell line (BxPC-3), demonstrating to be valid alternatives for the future development of this class of compounds. Previous works showed that aromatic linkers decreased the alkylation potency compared to aliphatic analogues. Since classical nitrogen mustards with milder electrophilicity are known to be also less toxic in vivo, we further expanded our library of compounds with a new and enlarged set of aromatic B-CePs. Interestingly, aromatic agents exerted a remarkable activity against 2D and 3D cancer cell cultures as well as an exclusive tropism for BxPC-3 cells, as observed for M-CePs. As a general rule, slower reactivity and enhanced transported-mediated uptake profiles were demonstrated to boost cytotoxicity against the more complex model of three-dimensional spheroids. The tropism for pancreatic adenocarcinoma BxPC-3 cells was further observed for a third set of Lys-ester derivatives bearing appended instead of embedded aromatic linkers. In this other study, an extended set of derivatives bearing Lys-ester/amide linkers with various aromatic and aliphatic side chains was investigated in terms of cellular activity and consumption kinetics due to ring hydroxylation in aqueous solution, a competing reaction which abates the fraction of compound able to alkylate the biological target. Our results highlighted that the lower DNA reactivity of Lys-ester aromatic analogues was compensated by a longer stability to hydroxylation, allowing them to preserve valuable indexes of cytotoxicity. In the last work presented here, we investigated the molecular bases of the observed 3-CePs tropism for the pancreatic adenocarcinoma cell line. Representative mono- and bifunctional compounds were thoroughly analyzed by the use of a multi-omic approach to investigate the molecular determinants of cell susceptibility to our drug candidates. We analyzed transcriptional changes and chromatin status upon treatment in a high- (pancreatic adenocarcinoma BxPC-3) and low-sensitive (colorectal adenocarcinoma HCT-15) cancer cell lines, demonstrating that BxPC-3 cells were unable to control proteostasis and DNA damage response in stress conditions when exposed to our alkylating agents. Furthermore, we derived perturbation-informed signatures predicting compound sensitivity and identified potentially more susceptible target tumor types for the further development of clinical candidates. We believe that our results exemplify the potential of a multi-omic approach in offering a versatile framework to support drug discovery toward precision oncology.
Tumor heterogeneity and complex resistance mechanisms undermine the therapeutic value of existing anticancer options, generating a pressing demand for alternative accessible and effective leads. In this sense, our group recently introduced 3-chloropiperidines (3-CePs) as a novel class of alkylating agents designed to improve the pharmacological profile of nitrogen mustard (NM) chemotherapeutics. Previous works described the improved alkylating properties of bifunctional 3-chloropiperidines (B-CePs) compared to the reference NM chlorambucil, exploring the effect of different linker structures on their reactivity with DNA and dissecting the mechanism of B-CePs alkylation at guanines. From these premises, this project further investigated the chemical space and antiproliferative value of these anticancer candidates. First, an explorative set of monofunctional analogues (M-CePs) was introduced to study the effect of a single reactive center on the activity of compounds. Notably, despite reduced DNA cleavage relative to the bifunctional compounds, these new agents showed nanomolar cytotoxicity indexes against test cancer cells, along with an unexpected tropism for a pancreatic adenocarcinoma cell line (BxPC-3), demonstrating to be valid alternatives for the future development of this class of compounds. Previous works showed that aromatic linkers decreased the alkylation potency compared to aliphatic analogues. Since classical nitrogen mustards with milder electrophilicity are known to be also less toxic in vivo, we further expanded our library of compounds with a new and enlarged set of aromatic B-CePs. Interestingly, aromatic agents exerted a remarkable activity against 2D and 3D cancer cell cultures as well as an exclusive tropism for BxPC-3 cells, as observed for M-CePs. As a general rule, slower reactivity and enhanced transported-mediated uptake profiles were demonstrated to boost cytotoxicity against the more complex model of three-dimensional spheroids. The tropism for pancreatic adenocarcinoma BxPC-3 cells was further observed for a third set of Lys-ester derivatives bearing appended instead of embedded aromatic linkers. In this other study, an extended set of derivatives bearing Lys-ester/amide linkers with various aromatic and aliphatic side chains was investigated in terms of cellular activity and consumption kinetics due to ring hydroxylation in aqueous solution, a competing reaction which abates the fraction of compound able to alkylate the biological target. Our results highlighted that the lower DNA reactivity of Lys-ester aromatic analogues was compensated by a longer stability to hydroxylation, allowing them to preserve valuable indexes of cytotoxicity. In the last work presented here, we investigated the molecular bases of the observed 3-CePs tropism for the pancreatic adenocarcinoma cell line. Representative mono- and bifunctional compounds were thoroughly analyzed by the use of a multi-omic approach to investigate the molecular determinants of cell susceptibility to our drug candidates. We analyzed transcriptional changes and chromatin status upon treatment in a high- (pancreatic adenocarcinoma BxPC-3) and low-sensitive (colorectal adenocarcinoma HCT-15) cancer cell lines, demonstrating that BxPC-3 cells were unable to control proteostasis and DNA damage response in stress conditions when exposed to our alkylating agents. Furthermore, we derived perturbation-informed signatures predicting compound sensitivity and identified potentially more susceptible target tumor types for the further development of clinical candidates. We believe that our results exemplify the potential of a multi-omic approach in offering a versatile framework to support drug discovery toward precision oncology.

Partendo da un'analisi preliminare dei processi improvvisativi che sono all'origine della musica Jazz, considero 3 diverse realtà organizzative del territorio veneziano che si occupano di promuovere e dare impulso a questo genere musicale: Veneto Jazz, Vicenza Jazz e Caligola, con una breve premessa introduttiva, in riferimento ad Umbria Jazz, la madre di tutti i Festival Jazz che si sono succeduti in Italia. L'obiettivo di studio dell'elaborato, è quello di mettere in evidenza per ognuna delle 3 diverse organizzazioni i seguenti elementi: i problemi organizzativi e di gestione, i problemi economico-finanziari, gli aspetti comunicativi e le ricadute extra-artistiche sul territorio (turismo e immagine), la considerazione sociologica (tipo di pubblico che accede a queste manifestazioni, il comportamento degli Enti locali nei confronti di queste organizzazioni artistiche (in che modo vengono finanziati i Festival Jazz in funzione delle politiche turistiche), la promozione dei musicisti italiani oltre ai già affermati musicisti americani.

L’epatocarcinoma (EC) è un tumore maligno ad elevata morbilità e mortalità la cui speranza di cura è basata su diagnosi precoce e terapia individualizzata. Ad oggi il trattamento di elezione è rappresentato dalla chirurgia e l’unico farmaco con comprovata efficacia terapeutica è un presidio biologico di recente introduzione ad azione anti-tirosinochinasica (Sorafenib). La gestione ottimale del paziente con EC è sempre più influenzata dalla possibilità di disporre di marcatori molecolari da utilizzarsi nella pratica clinica a significato diagnostico, prognostico e predittivo. Il presente lavoro si propone come contributo all’identificazione e validazione di specifici indicatori biologici nella diagnostica del piccolo EC, predizione della recidiva del tumore resecato e della sensibilità individuale alla risposta al trattamento farmacologico nella forma avanzata. E già stato dimostrato che l’accuratezza della diagnosi precoce di EC su biopsia epatica puo’ essere migliorata in maniera significativa dalla valuatazione dell’espressione combinata di alcuni marcatori recentmente proposti e già utilizzati nella pratica clinica. Abbiamo cercato di ottimizzare questo pannello valutando il contributo aggiuntivo della catena pesante della clatrina (CHC) che è componente essenziale nel sistema di trasporto vescicolare intra ed extracellulare, già segnalato come sovraregolato negli epatociti tumorali maligni. Utilizzando una casistica di EC di piccole dimensioni e di lesioni precancerose di controllo e documentando l’espressione della molecola con tecnica immunocitochimica, abbiamo verificato la sua efficacia in termini di accuratezza diagnostica. In particolare l’introduzione di CHC si è dimostrata in grado di incrementare la sensibilità del pannello dal 46,8% al 63,8% a fronte del mantenimento di una specificità assoluta. E’ noto come il processo di carcinogenesi epatica origini anche sulla base di alterazioni delle vie molecolari che governano i processi di tipo ossidativo, con importante ruolo di protezione svolto dalla famiglia dei citocromi. Alcuni studi di espressione genica hanno già evidenziato che la recidiva di malattia tumorale nel fegato sia influenzata dalla disregolazione di alcuni citocromi, tra cui CYP1A2. Abbiamo inteso verificare e validare il ruolo del citocromo CYP1A2 nella qualità di possibile marcatore ad impatto traslazionale nella recidiva dell’EC attraverso il suo dosaggio immuncitochimico nel parenchima epatico non tumorale. Utilizzando una casistica omogenea di pazienti portatori di EC ad eziologia virale (virus C dell’epatite) e a follow-up noto, il dosaggio di CYP1A2 è risultato associato in maniera statisticamente significativa, sia in analisi univariata che multivariata al tempo libero da recidiva tumorale (p=0.012). In particolare la ridotta espressione del citocromo negli epatociti non tumorali, interpretabile come background più suscettibile alla trasformazione neoplastica “de novo”, risultava in grado di predire una più rapida ripresa della malattia. La terapia farmacologica dell’EC è attualmente somministrata a pazienti portatori di forme avanzate senza tenere conto della reale e individuale suscettibilità all’azione del farmaco. Vi è una forte esigenza clinica di selezionare i pazienti candidabili al trattamento anche per gli elevati costi della terapia, ma nessun marcatore, sia esso tissutale o sierologico, si è rivelato ad oggi capace di soddisfare questi requisiti. Abbiamo inteso esplorare se i miRNA, che costituiscono una famiglia eterogenea ma fondamentale di molecole regolatrici della traduzione, potessero a loro volta candidarsi a possibili marcatori biologici di predizione della sensibilità al trattamento farmacologico dell’EC. A questo scopo, partendo da un pool amplissimo di miRNA, abbiamo proceduto ad una loro progressiva selezione sulla base della loro disregolazione nonchè associazione con il tempo di progressione della malattia dopo trattamento. Lo studio è stato condotto su biopsie epatiche pretrattamento di EC di pazienti omogeneamente trattati con Sorafenib. La selezione delle molecole e la validazione delle stesse è stata condotta in due casistiche distinte di cui la seconda utilizzata come coorte di validazione. Abbiamo cosi potuto individuare un unico miRNA con meccanismo di azione e geni targets poco conosciuti, il cui sovradosaggio valutato con tecnica di real-time PCR risultava fortemente e statisticamente associato ad una prolungata stabilizzazione della malattia tumorale (p=0.009). L’insieme di questi risultati dimostra che la crescente domanda di marcatori di medicina personalizzata per ottimizzare la gestione del paziente oncologico (con EC nel nostro caso), possa essere soddisfatta attraverso l’applicazione di tecniche relativamente semplici e riproducibili quali l’immunocitochimica e l’analisi quantitativa del messaggero, condotte sia su tessuti tumorali che non tumorali. Naturalmente affinchè il dosaggio di una molecola possa entrare nella pratica clinica corrente, candidandosi come un autentico marcatore molecolare, è necessaria, come nei nostri casi, un’ulteriore validazione in studi di tipo prospettico.
Hepatocellular carcinoma (HCC) is a malignant tumor with high morbidity and mortality worldwide. The treatment is based on early diagnosis and individualized therapy. Today surgery is the primary strategy for patients with HCC and Sorafenib, an anti tyrosine kinase drug, represents the only available drug with proven efficacy. The discovery of new molecular markers (diagnostic, prognostic and predictive) that can be translate in the clinical practice represents the improvement in the management of these patients. The present research is aimed at identifying and validating biological markers related to the early HCC diagnosis and to predict HCC relapse. In addition the individual sensibility to pharmacological treatment in patients with advanced HCC is also evaluated. It has been shown that the accuracy of the HCC early diagnosis on the liver biopsy can be significantly improved by combining histological features with molecular markers, recently proposed and applied in the clinical practice. We have optimized this molecular panel for evaluating the contribution of the clathrin heavy chain (CHC), which is an essential component of intra and extracellular vesicular transport, and previously reported as an up-regulated molecule in malignant liver cells. Using a series of small HCC and control precancerous lesions we tested, by immunohistochemical analysis, whether CHC expression contribute to improve the histopathological diagnostic accuracy. We found that the introduction of CHC increases the sensitivity from 46,8% to 63,8% and maintains an absolute specificity. It is now ascertained that hepatocarcinogenesis is based on alterations in a wide variety of molecular pathways controlling oxidative processes and that cytochromes family plays a protective role in this context. Additionally, gene expression profiles have shown that relapse of HCC is caused by some cytochromes deregulations (i.e. CYP1A2). We verified and validated the CYP1A2 role in relation to relapse of disease by immunohistochemistry and its quantification in non-tumoral liver parenchyma. By investigating a homogeneous series of HCC patients with HCV infection and known follow-up, we demonstrated that the CYP1A2 dosage was associated with the recurrence-free survival (p=0,012) in both univariate and multivariate analyses. Moreover, CYP1A2 expression decreased in non-tumoral hepatocytes, the most susceptible background to “de novo” neoplastic transformation, predicted a faster recurrence of disease. The pharmacological therapy of HCC is currently administered to patients with advanced disease, without any discrimination according to the individual drug susceptibility. There is a compelling clinical need to select patients more suitable to underwent pharmacological treatment, because of the elevated cost. Furthemore, there is no still histological and/or serological biomarker, capable to satisfy these requirements. For this reason, we explored whether miRNA, heterogeneous family of regulatory molecules of translation, can be used as new molecular predictive markers of sensitivity to HCC treatment. For this purpose, starting from a wide pool of miRNA, we progressively selected those deregulated and associated with time to progression of disease after treatment. This research was conducted on liver biopsies sampled before treatment in patients treated with Sorafenib. The selection and validation of miRNA were conducted on two distinct series of patients. Among these the second group of patients has been used as a validation set. We identified a single miRNA, with mechanism of action ad genes target unknown, which when appears overexpressed, by real-time PCR analysis, was strongly and statistically associated with prolonged stabilization of tumor disease (p=0,009). The above findings all suggest that the increasing demand for customized markers to optimized the management of HCC patients can be investigated through the application of relatively simple and reproducibly techniques (including immunohistochemistry and real-time PCR) on both tumoral and non-tumoral tissue. Further validation in prospective studies is required to pursue a personalized dosage of new molecular markers to be introduced in the current clinical practice.

L’emersione di nuove scoperte scientifiche e lo sviluppo di nuove tecnologie hanno portato a un ripensamento del tema della prova scientifica nel processo penale in prospettiva interdisciplinare. Se, da un lato, la ricostruzione probatoria dei fatti di reato è sempre più affidata ai risultati della prova tecnico-scientifica, dall’altro agli strumenti tradizionalmente noti se ne stanno affiancando di nuovi, la cui affidabilità e rilevanza è spesso controversa. La presente ricerca nasce dall’interesse verso il recente affacciarsi nel panorama giuridico di strumenti e prospettive scientifiche relativamente nuove, come le neuroscienze cognitive e le tecniche a esse collegate. Se, da un lato, vi è un grande interesse da parte di giuristi e scienziati ad approfondire il tema, dall’altro però l’indagine è ancora disorganica, dispersa tra tematiche diverse, con profili di approfondimento che mutano anche a seconda della prospettiva che si assume: nazionale o internazionale, giuridica o multidisciplinare. L’ambito di riferimento del presente lavoro, è quello dell’utilizzo delle neurotecniche nel processo penale. Si possono rilevare diversi piani sui quali si può condurre l’analisi: a) Un piano di tipo penale-sostanziale, con riferimento all’accertamento dell’imputabilità; b) Un piano di tipo procedurale, con riferimento all’ammissione della prova scientifica nel processo c) Un piano di tipo costituzionale, con riferimento alla protezione dei diritti fondamentali dell’imputato. Questi tre livelli sono, in realtà, strettamente interconnessi e la libertà personale è il presupposto e il punto di arrivo dell’indagine penale: la sua protezione o compressione costituiscono il fil rouge di tutti i piani di analisi, dal momento che la custodia cautelare e la pena detentiva sono una violazione, pur giustificata da esigenze di giustizia e di prevenzione, ma pur sempre violazione, della libertà personale di un essere umano. E va notato come, una volta che si sia assunta l’idea ampia di libertà personale che le corti costituzionali ormai hanno esplicitamente affermato (in Italia, si pensi alla sentenza n. 471 del 1990), potenziali altre violazioni ad essa, nella sua dimensione psicofisica, si possono riscontrare in altri momenti e per altre parti del processo, come per esempio, con riferimento alle modalità di escussione del teste /acquisizione della prova nel corso del processo. Da ciò discende la centralità, nel valutare il modo in cui alcune recenti scoperte scientifiche e le tecniche connesse possano incidere sul diritto penale, della protezione della libertà personale dell ’imputato e delle parti del processo, e di come essa sia regolata nei diversi sistemi e posta in evidenza nella casistica giudiziaria. Queste considerazioni sono centrali nella presente ricerca: partendo dal fatto che la libertà personale costituisce un presupposto condiviso nei contemporanei sistemi giuridici occidentali, si è mirato a osservare il modo in cui si atteggia il rapporto tra tale libertà dell’imputato (e delle altre parti nel processo) e il potere autoritativo dello stato nel processo penale, laddove le nuove tecniche neuroscientifiche hanno applicazione. L’analisi è stata condotta in una prospettiva comparata tra il sistema giuridicocostituzionale italiano e quello statunitense.

2012 - 2013
The aim of this Doctoral work is double: (a) to offer the complete edition of three important distinctiones – the second, the third and the eighth – coming from the Sentence Commentary of the Franciscan John of Ripa, who lectured in Paris in the period 1354-1358. These three distinctiones constitute a homogeneous group of questions dealing with the metaphysical issues of the existence and nature of God (immensitas) in its distinction from the nature of created beings (infinitas/finitas), together with the original mechanism called replicatio unitatis divinae which regulates the generation of all created beings [Distinctio 2], the analogy between God and creatures, with the lucid restriction – consciously elaborated after Duns Scotus – of the univocity in the sole created domain [Distinctio 3], and the formal distinction between all the perfectiones originally contained in the divine essence, which do not introduce any form of multiplicity in the Unity of the supersimplex (immensa) divine essence [Distinctio 8]; (b) to offer an elaborated study of the themes present in these distinctiones, not only regarding to Ripa’s thought, but also through the analysis of related philosophical perspectives (Hugolinus of Orvieto, Pierre Ceffons, John of Mirecourt, Francis de Mayronis, Francis of Appignano, Gregory of Rimini and William of Ockham). This duplex intent is reflected in the two volumes which compose this thesis, which aims to discover the thought of a long-time underestimated philosopher. [edited by author]
XII n.s.

LA TESI MIRA A SPIEGARE DUE PROCESSI COGNITIVI QUALI: L'ANALOGICAL THINKING E IL CONCEPTUAL BLENDING ATTRAVERSO UNA PROSPETTIVA TEORICA E UNA CONSEGUENTE ESPLICAZIONE ATTRAVERSO DUE CASE STUDIES UNO RIGUARDANTE L'OROLOGIO CIFRA 3 DELL'AZIENDA SOLARI E L'ALTRO RELATIVO ALLA MAPPA DELLA METROPOLITANA

With the law August 3, 2007, n. 124 entitled "Information system for the security of the Republic and the new discipline of the secret", as amended by the Law of 7 August 2012, n. 133, the state legislature has completely changed the system of national security, thirty years after his first discipline. What were the reasons for this change? Why has Parliament, in this context, decided to create a new institutional architecture? And above all, was it really offer the maximum possible protection to the value of the salus rei publicae, in the light and the limits of the Constitution? The present study, through a survey of the origins, constitutional compatibility and dynamics of law n. 124 of 2007 aims to provide answers to these problems. In this regard will be examined parliamentary proceedings leading to the adoption of this law and its further editing; will be identified all the issues of constitutional and political institutions emerging from the renewed discipline; will be study the main decisions of the Constitutional Court on the matter. At the end will be also analyzed certain judgments of the European Court of Human Rights, for two reasons: because the law can be an inspiration to the legislature in future unavoidable revisions of the subject and because the Constitutional Court may, by looking at the judge Strasbourg, go back to being the real judge of a state secret. Law no. 124 of 2007, as amended in 2012, is not an unconstitutional law but the original purpose of the legislature to create a system of salus rei publicae compatible with the balance of the parliamentary form of government has failed and that, above all, because of inadequate counter-seater ever after confirmation of the secret, and, therefore, when damage is possible occurred. So, who does control the head of government, which is the only dominus of salus rei publicae? Nobody.

GATA-3 è posto nel braccio corto del cromosoma 10 ed è un membro della famiglia di fattori di trascrizione GATA(1-6) che legano specifiche sequenze GATA. GATA-3 è coinvolto nella modulazione dell'espressione genica durante l'embriogenesi di molti apparati umani e delle cellule ematopoietiche e risulta altamente conservato nei vertebrati. La delezione terminale del cromosoma 10 causa una sindrome clinica chiamata sindrome HDR caratterizzata da ipoparatiroidismo, malformazioni cardiache e del sistema genito-urinario, immunodeficit e ipoacusia. Nel nostro lavoro abbiamo voluto analizzare l'espressione della proteina GATA3 tramite immunoistochimica (GATA3, Pharmigen, L50-823, 1:150) in una ampia casistica di tessuti normali e neoplastici nell'apparato urinario, della mammella e di tiroide e paratiroide. Abbiamo evidenziato che il GATA3 è espresso nell' urotelio normale e neoplastico della pelvi renale 78%) e della vescica (88%) e l'espressione è correlata al grado di differenziazione delle neoplasie. Nel rene abbiamo notato espressione nei podociti, nel dotto collettore e nei tubuli distali, mentre nella controparte neoplastiche quasi tutte le neoplasie studiate sono risultate negative tranne 4 casi di carcinoma a cellule chiare papillare e 5 casi di carcinoma a cellule chiare con prominente proliferazione leiomuscolare. I casi risultati positivi all'immunoistochimica sono stati poi confermati mediante Western-blot. Abbiamo inoltra valutato il gene VHL dei casi di carcinoma a cellule chiare papillare notando che nessun caso portava mutazioni o metilazione del gene VHL. Nella mammella abbiamo confermato i dati di letteratura che vedono il GATA3 correlare a tumori meglio differenziati e a prognosi benigna (basso grado citologico, basso indice proliferativo, espressione di ER e PgR) ma abbiamo anche evidenziato un'inaspettata positività al marcatore nei gruppo di casi tripli negativi (ER neg, Prg neg e HER 2neg). Dato che il GATA3 risulta fortemente associato all'espressione dei recettori per estrogeni, la nostra scoperta permetterebbe di individuare un sottogruppo di pazienti che potrebbero beneficiare della terapia anti-estrogenica. Il GATA3 risulta importante anche nello sviluppo degli organi endocrini del collo, nel nostro studio abbiamo evidenziato una positività del GATA3 nei solid nest della tiroide, nel parenchima normale ed in quello adenomatoso delle paratiroidi ed è risultato positivo anche in un citologico di adenoma paratiroideo, sottolineando il possibile uso in citologia per la diagnostica differenziale tra le lesioni paratiroidee (GATA3+) e quelle follicolari tiroidee (GATA3 neg).
GATA-3 (10p 14-15) is a member of transcription factor family (GATA1-6) that binds selected GATA sites to regulate specific gene expression. GATA-3 is involved in the modulation of gene expression profiles during embryogenesis of a variety of human tissues as hematopoietic cells Terminal deletions of chromosome 10p result in a DiGeorge-like fenotype characterized by hypoparathyroidism, heart defects, immune deficiecy, deafness and Renal malformation (HDR syndrome). We sought to evaluate the presence of GATA-3 protein by immunohistochemistry among a larger cohort of subtypes of carcinomas and normal tissue of urogenital tract, breast, thyroid and parathyroid, these cases were identified and retrieved from the archives of Verona University and diagnosed between 1993 to 2011. Our series includes: 86 urothelial carcinoma, 23 metastatic localization of urothelial carcinomas, 100 renal cell neoplasms, 20 prostatic adenocarcinoma, 133 normal tissue, 191 breast carcinomas, 1 citologic smear from parathyroid adenoma, 10 non-neoplastic parathyroid parenchima, 10 parathyroid adenomas, 10 solid nests in the thyroid, 10 papillary thyroid carcinoma, 5 follicular thyroid carcinomas, 20 bening hyperplastic nodules and 5 brachial cysts. We preformed an immunohistochemistry staining with mouse monoclonal anti-GATA3 was obtained from Pharmigen (L50-823, dilution 1:150). The pattern of staining was nuclear. We assesed Western blot in renal cell neoplasms wich are GATA-3 positive and in some of these cases VHL sequencing analysis and CpG methylation analysis. Regarding breast cases we had collected personal data and histological diagnosis for each case, wich include: diameter, grade, lymphonode status, estrogen receptor alpha (ER), progesterone receptor (PR), proliferation index (Ki67), HER-2. In normal kidney tissue GATA3 stained podocytes, the epithelium of distal nephron and also collector ducts; bladder urothelial epithelium showed strong GATA3 nuclear positivity; GATA3 stained some prostatic basal cells of glandular structures and basal cell of seminale vescicles. Pelvis urothelial carcinomas were positive in 78% of cases (25/32), furthermore there was evidence that tumors with good-moderate differentiation (88%) were more positive those poor-differentiated (71%). Bladder urothelial carcinomas were positive in 85% of cases (18/21) with correlation from GATA3 expression and differentiation grade of neoplasms (100% low grade vs. 75% high grade). Bladder carcinomas nested variant and micropapillary variant showed GATA3 positivity (1/1). All prostatic localizations of bladder carcinoma had showed positive immunoreaction for GATA3 (6/6). Urothelial metastatic localizations were positive in 57% of case (13/23). None prostatic adenocarcinoma swowed GATA3 positivity. In renal neoplasms we have found GATA-3 positivity in 4 of 5 clear cell-papillary renal cell carcinoma and in 5/5 clear renal cell carcinomas with prominent leiomyomatous proliferation. None other cases of renal carcinomas showed positivity to GATA-3. In 2 clear cell-papillary renal cell carcinoma and in 1 clear renal cell carcinomas with prominent leiomyomatous proliferation Western blot analysis highlighted the presence of the GATA-3 protein in cases positive at immunophenotyical level. No mutation of coding sequence of the VHL gene was found in epithelial and smooth muscle cell components of the tumors . MS-MLPA analysis of FFPE tissue DNA samples showed absent or mild methylation of VHL gene. In normal breast tissue GATA-3 stained duct luminal cell. Overall 147 breast cancer (78%) have shown GATA-3 expression. We have found statistically significance correlation from GATA-3 expression and patient age, grading of neoplasm, ER-PgR status and Ki-67 expression. In our collection all 106 ER+, PG, breast carcinoma were positive for GATA-3. Among ER-,PG-, Her2+ breast carcinoma the presence of GATA-3 was detected in 10/11 cases The most interesting and unexpected evidence of our work is GATA-3 expression in triple 31 cases of negative breast carcinoma (31/74)(41,9%). We observed strong positivity in cytologic sample of parathyroid adenoma, in all solid nests (10/10), parathyroid parenchymas (10/10), parathyroid adenomas (10/10). and in the epithelial lining of branchial cysts (5/5). On the contrary, GATA3 immunostaining was absent in all papillary (0/10) and follicular thyroid carcinomas (0/5) and in all benign hyperplastic thyroid nodules (0/20). Finally this study had highlighted that GATA3 is a good marker to reveal urothelial origin of neoplastic cells and allow to use GATA3 in the differential diagnosis between prostate carcinomas and urothelial tumors; moreover we found that urothelial tumors tend to lose GATA3 expression in low differentiated areas. Among renal cell neoplasms, we interestingly noted that only rare tumours such as clear cell-papillary renal cell carcinomas (4/5) and clear cell renal cell carcinomas with prominent leiomyomatous proliferation (5/5) were positive for GATA-3. GATA3 protein expression is confirmed by using the Western Blot analysis. In our study we found that: clear cell papillary renal cell carcinoma do not show VHL mutation and do not show CpG methylation abnormalities and may be part of this tumors subgroup lacking VHL gene alterations (WT VHL) as indicated in the recent study. Some studies are supporting the evidence that under the name “clear cell renal cell carcinoma” are included a basket of different entities. These rare cancers are difficult to diagnosis to inexperienced pathologist and in the literature reliable data for specific markers for these neoplasm are lacking. So GATA3 antibody merits to be included in the most common routinely available immunohistochemical panels and to be used for the differential diagnosis of renal cell neoplasms. Breast cancer comprises a heterogeneous group of diseases that vary in morphology, biology, behaviour and response to therapy. In our series in normal breast tissue GATA-3 stained duct luminal cell, as previously validated . Overall 147 breast cancer (78%) have showed GATA-3 expression and we demontrated GATA-3 expression statistically is correlated with patient age, ER-PgR status, grading of neoplasm and Ki-67 expression. We found that all 106 ER+/PG+, breast carcinoma were positive for GATA-3. Among ER-/PG-/Her2+ breast carcinoma the presence of GATA-3 was detected in 10/11 cases The most interesting and unexpected evidence of our work is GATA-3 expression in triple negative breast carcinoma (31/74)(41,9%). This percentage of triple negative tumors with positive GATA3 may highlight a new subgroup of tumors that benefit therapy with tamoxifen. In our series we observed strong positivity in all solid nests, parathyroid parenchymas, parathyroid adenomas and in the epithelial lining of branchial cysts. On the contrary, GATA3 immunostaining was absent in all papillary and follicular thyroid carcinomas and in all benign hyperplastic thyroid nodules. A specific use of GATA3 could be in fine needle aspiration (FNA) cytology of parathyroid lesions. No one single cytomorphologic feature is diagnostic and at the present we can use nucler antibod, TTF1, to recognise follicular lesion and a citoplasmatic antibody, PTH, to recognise parathyroid lesion. But in citologic smear it is easy that cells losing the cytoplasmic component and the diagnosis of parathyroid adenoma is possible only in a negative manner. GATA3 is a nucler marker and it can do very useful to identify parathyroid cells.

Synthesis and biological evaluation of new saccharin-based inhibitors of cancer-related carbonic anhydrase IX and XII isoforms Carbonic anhydrases (CAs, EC 4.2.1.1) are widespread metalloenzymes implicated in many cellular and physiological processes requiring bicarbonate as substrate (e.g. electrolyte secretion, pH homoeostasis, respiration etc.). The fifteen human isoforms of carbonic anhydrase (hCAs) differ in cellular localization (cytosol, mitochondria or cell membrane), sensibility to inhibitors and catalytic activity. hCAs are well established therapeutic targets to treat a wide range of disorders. In particular, hCA IX and XII have been recognized as tumor-related isoforms, participating in the complex machinery which regulate the pH of cells in hypoxic tumors. In our previously reported studies we showed that N-alkylated saccharin derivatives effectively inhibited the two cancer-related isoforms. Here we report the design, synthesis and the inhibitory activity of novel compounds based on saccharin scaffold. We used two different approaches in order to obtain two series of new compounds. In the first approach, we performed a reductive ring opening of N-alkylated saccharin derivatives, obtaining molecules endowed with secondary sulfonamide and primary alcohol. In the second approach, a series of tertiary sulfonamides (saccharin/isoxazole and saccharin/isoxazoline derivatives) were obtained by the insertion of isoxazole or isoxazoline moiety as spacer between the saccharin nitrogen and (un)substituted aryl or heteroaryl rings. All the synthesized compounds were tested to evaluate their inhibitory activity towards the ubiquitous off-target isoforms, hCA I and II, and the cancer-related ones, hCA IX and XII, by a stopped-flow, CO2 hydrase assay. All the compounds were inactive against the two cytosolic off-target hCA I and II (Kis > 10 µM), while showed inhibitory activity in the low nanomolar range against the two cancer-related isoforms hCA IX and XII. Computational approach has been performed in order to better understand the binding mode of these inhibitors. Benzo[b]tiophen-3-ol derivatives as effective inhibitors of hMAOs: design, synthesis and biological activity Monoamine oxidases (MAOs; EC 1.4.3.4) are mitochondrial bound flavoenzymes, which catalyse the oxidative degradation of amines. They have been widely recognised as important pharmacological targets for the treatment of mood disorders (anxiety, depression) and neurodegeneration (Parkinson’s disease, PD) as the result of their effects on monoamine metabolism and level. Furthermore, evidences show the probable implication of these enzymes in some tumors (e.g prostatic cancer) and cardiomyopathies. Based on tissue localization, structural homology, active site differences, substrate/inhibitor selectivity and catalytic efficiency, two isoforms have been characterized (MAO-A and MAO-B). Both of them could be the target of selective inhibitors acting as reversible or irreversible agents. An extensive number of natural and synthetic compounds have shown effective inhibition of human MAOs. Indole analogues, aurones and indanone derivatives are examples of MAO inhibitors. They show a common structural feature that is the chalcone moiety, whose ability to bind hMAO enzymes has been reported by our research group. With the aim to expand our knowledge about hMAO inhibtion, we performed the design, synthesis, characterization and in vitro biological activity evaluation of some novel benzo[b]thiophen-3-ol derivatives. This scaffold retained the above cited chalcone system, differing from the already studied inhibitors in some features like the isosteric replacement of aurones’ oxygen with sulphur one, the presence of 1,3-diketone system giving keto-enol tautomerism and the presence of a carbonyl group instead of methylylidene one, bound at position 2 of the bicyclic system. These compounds have been synthesised through a new simple synthetic approach consisting in a one-step reaction, which led to moderate to high yields. The obtained compounds showed activity in the micromolar/low micromolar range against hMAO-B with the best inhibition profile observed for compounds containing phenyl ring substituted in the meta- position with halogens. These compounds have also been tested with cortex synaptosomes in both basal and LPS-induced inflammatory conditions to evaluate their ability to affect the DOPAC/DA ratio as well as LDH activity. DOPAC/DA ratio is an indirect index of MAO-B activity, and all the tested compounds are effective in reducing this value in cortex synaptosomes challenged with LPS, showing outcomes better than the reference drug deprenyl. Furthermore, all the tested molecules inhibited LDH activity in the concentration range 0.1-1 µM, showing potential activity as neuroprotective agents.

Respiratory diseases are a major cause of human morbidity and mortality and are a leading cause of economic loss to livestock producers. The respiratory tract is constantly in contact with dust, bacteria, fungi, viruses and other pathogenic agents that are found in the air. Normally, the body has the ability to clear these foreign particles. However, physiological and environmental stresses can impair airway defense mechanisms resulting in establishment of pulmonary infections. The microbes and their products engage various receptors in the lung to activate epithelium, endothelium, macrophages, neutrophils and other cells. The activation of inflammatory cascade in the lung results in recruitment of neutrophils, damage to air-blood barrier and development of edema. Although there have been significant advances in our understanding of mechanisms of lung inflammation, there have been a lack of any significant advances in the development of new therapeutics to manage lung disease, which may suggest that our understanding of the inflammatory mechanisms is still incomplete. Pentraxin 3 (PTX3) is an innate immune protein which has been implicated in a diverse range of inflammatory processes, such as recruitment of cells and production of cytokines. PTX3 is an acute phase protein, with low or undetectable levels in the circulation of healthy humans and animals, and rapid, dramatic increase in inflammatory diseases. The expression and function of this protein has not been characterized in the lungs of domestic animal species. Because of potential implications of PTX3 in lung inflammation, I studied the expression of PTX3 in normal and inflamed lungs of calves, pigs, horses, foals and humans. Lungs from all of these species showed expression of PTX3 in airway epithelium, alveolar septa, vascular endothelium and inflammatory cells. Western blot performed on homogenates from normal and inflamed lungs from calves and pigs show an increased expression of PTX3 in inflamed lungs (P<0.05). Because protein function is influenced by its location in the cell, I clarified the subcellular expression of PTX3 with immuno-electron microscopy on normal and inflamed calf and horse lungs. PTX3 was localized on pulmonary intravascular macrophages, monocytes, neutrophils and, unexpectedly, platelets. PTX3 was also present in the nuclei of neutrophils, monocytes and pulmonary intravascular macrophages. Neutrophils are critical regulators of acute lung inflammation. Having observed PTX3 in neutrophils, I investigated the effect of E. coli lipopolysaccharide-induced activation on PTX3 in neutrophils in vitro. Neutrophils challenged with E. coli LPS were examined at 30, 60, 90 and 120 minutes after the treatment. Normal peripheral blood neutrophils showed PTX3 expression. Neutrophils activated with LPS appeared ruffled and showed loss of PTX3 expression at 30 minutes followed by recovery of the expression. Western blots performed on normal and activated neutrophil homogenates did not show any differences (P=0.05). Collectively, the data show PTX3 in normal and inflamed lungs across multiple species. PTX3 was also detected in normal and activated neutrophils. While the function of intriguing localization of PTX3 in the nuclei as well as in platelets is not known, the similarity of expression across the species suggest a role for PTX3 in lung inflammation.

Bovine adenovirus (BAdV)-3 is a non-enveloped icosahedral DNA virus, which replicates in the nucleus of infected cells, and is being developed as a vector for vaccination for humans and animals. The genome of BAdV-3 is organized into early, intermediate and late genes and it has thirty three predicted open reading frames (Reddy et al., 1998). The late region of BAdV-3 is divided into seven families (L1-L7) (Reddy et al., 1998). One of the proteins expressed in the L-6 region encodes a protein called pVIII, which is a minor capsid protein connecting the core with the inner surface of the capsid. The objective of the current study was to characterize pVIII protein of BAdV-3 and to examine its role in the life cycle of BAdV-3. Anti-pVIII serum detected a protein of 24 kDa at 12-48 hr post infection and an additional protein of 8 kDa at 24-48 hr post infection. While a 24 kDa protein is detected in empty capsids, only the C-terminal cleaved protein of 8 kDa is detected in the mature virion suggesting that amino acids 147-216 of conserved C- terminus of BAdV-3 pVIII are incorporated in mature virions. The pVIII protein predominantly localizes to the nucleus of BAdV-3 infected cells utilizing the classical importin α /β dependent nuclear import pathway. Analysis of mutant pVIII demonstrated that amino acids 57-72 of the conserved N-terminus bind to importin α-3 with high affinity and are required for the nuclear localization. Detection of hexon associated with both, precursor (24 kDa) and cleaved (8 kDa) form of pVIII suggests that the C-terminus of pVIII interacts with Hexon. Based on yeast II hybrid screening assay, we identified the cellular protein DDX3 as an interacting protein partner of pVIII. Earlier, targeting of DDX3 by few viral proteins has defined its role in mRNA transport (Yedavalli et al., 2004) and induction of interferon production (Schroder et al., 2008; Wang et al., 2009). Here, we provide evidence regarding the involvement of DDX3 in cap dependent cellular mRNA translation and show that targeting of DDX3 by the adenovirus pVIII protein abolishes cap-dependent mRNA translation function of DDX3 in virus infected cells. Adenovirus late protein pVIII interacts with DDX3 in transfected and bovine adenovirus (BAdV-3) infected cells. pVIII inhibited capped mRNA translation in-vitro and in-vivo by limiting the amount of DDX3 and eIF3. Diminished amount of DDX3 and eIFs including eIF3, eIF4E and PABP were present in cap binding complex in BAdV-3 infected or pVIII transfected cells with no trace of pVIII in the cap binding complex. The total amount of eIFs appeared similar in uninfected or BAdV-3 infected cells. The co-immunoprecipitation experiments indicated the absence of direct interaction between pVIII and eIF3, eIF4E or PABP. These data indicate that interaction of pVIII with DDX3 depletes eIF3, eIF4E and PABP from the cap-binding complex. We conclude that DDX3 promotes cap-dependent cellular mRNA translation and BAdV-3 pVIII inhibits translation of capped cellular mRNA by excluding functional cap-binding complex from the capped cellular mRNA. BAdV-3 infection of DDX3 positive cells significantly inhibits cellular protein synthesis at late times post-infection. Interestingly, knockdown of DDX3 resulted in significant reduction in virus yield and expression of BAdV-3 late proteins at late times post-infection. Our results suggest that selective translation of BAdV-3 late mRNAs observed at late time post-infection of DDX3 positive cells is abrogated in DDX3 knock down cells. Moreover, the reduction in the extent of protein synthesis is evidenced by less functional 80S and polysomes in pVIII expressing plasmid transfected cells. Alternatively, DDX3 and pVIII binds to BAdV-3 tripartite leader (TPL) and the translation of mRNAs containing TPL at their 5’ ends is enhanced in the presence of pVIII and DDX3 proteins. From this observation, we concluded that pVIII and DDX-3 might promote the translation of late viral mRNAs by interacting with TPL.

This dissertation describes the approach taken in measuring two neutrino double beta decay of Cd-116 to the ground state of Sn-116 and in searching for the effective Majorana neutrino mass by placing a lower limit on the half-life for neutrinoless double beta decay of Cd-116 using the powerful technique of a combined tracking chamber and calorimeter with the NEMO-3 detector. The description of the detector, its natural background contamination, and the tools used to perform the analysis are discussed. The single electron channel was used to identify source foil contamination from [beta]-emitters and the electron-gamma channel was used to confirm the previous measurements of Tl-208 and Bi-214 contaminations in the source foil. Using these backgrounds, the two neutrino double beta decay half-life of Cd-116 was measured for the single states dominance hypothesis and the higher states dominance hypothesis. The final data set was defined to be data from Phrase One and Phase Two for the medium and low activity regions. Using 1471 days of data, the values of the half-life for the single data dominance hypothesis and the higher states dominance hypothesis were found.
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Most LEED (Leadership in Energy and Environmental Design) certified buildings are commercial office buildings and multi-use buildings. As of October 2009, 35,000 projects were registered in the LEED system, "comprising over 4.5 billion square feet of construction space in all 50 states and 91 countries." However, as of April 30, 2009, only 43 healthcare projects had achieved LEED certification. Currently, most studies focus on the economic benefits and energy consumption of LEED certified buildings, rather than human factors. A small gain in productivity can result in a heftier financial gain. Even modest improvements in productivity and absenteeism can substantially outweigh the energy cost. This study surveyed 164 staff in the two healthcare settings for case study, and 146 staff in the six LEED certified healthcare settings for the main study in climate zone 2 and 3. Telephone interviews with the six facility managers were used to verify the survey results and further examine the healthcare facilities? performance and the effectiveness of the LEED strategies from facility managers' perspectives. Independent t-test was used to examine the difference between the LEED and Non-LEED hospitals in one healthcare system and results showed that building performance were rated higher by staff in LEED certified hospital than Non-LEED hospital. MANOVA was conducted to compare the staff's ratings between Silver and Gold certification levels, male and female, and also explore the possibility of interaction effect. Multilevel regression modeling was used to test how the building performance variables affect the overall comfort and productivity. Study results showed that staff in the Gold certified hospital had significant higher ratings in most the performance variables. Gold certified healthcare settings were significant better in rated building overall, overall comfort and controllability than Silver certified healthcare settings. And males felt more comfortable in temperature than females in healthcare facilities. Regarding the overall comfort and productivity, building design, efficiency of the space use, temperature comfort and controllability over building system were significant predictors for staff overall comfort; and lighting comfort, temperature comfort and controllability over building system had significant positive relationship with perceived productivity. LEED certified healthcare settings appear to have a good environment and building performance for occupants. Controllability, lighting, temperature, use of space, building design were important factors in staff comfort and productivity.

Technology scaling of CMOS devices has made the integrated circuits vulnerable to single event radiation effects. Scaling of CMOS Static RAM (SRAM) has led to denser packing architectures by reducing the size and spacing of diffusion nodes. However, this trend has led to the increase in charge collection and sharing effects between devices during an ion strike, making the circuit even more vulnerable to a specific single event effect called the single event multiple-node upset (SEMU). In nanometer technologies, SEMU can easily disrupt the data stored in the memory and can be more hazardous than a single event single-node upset. During the last decade, most of the research efforts were mainly focused on improving the single event single-node upset tolerance of SRAM cells by using novel circuit techniques, but recent studies relating to angular radiation sensitivity has revealed the importance of SEMU and Multi Bit Upset (MBU) tolerance for SRAM cells. The research focuses on improving SEMU tolerance of CMOS SRAM cells by using novel circuit and layout level techniques. A novel SRAM cell circuit & layout technique is proposed to improve the SEMU tolerance of 6T SRAM cells with decreasing feature size, making it an ideal candidate for future technologies. The layout is based on strategically positioning diffusion nodes in such a way as to provide charge cancellation among nodes during SEMU radiation strikes, instead of charge build-up. The new design & layout technique can improve the SEMU tolerance levels by up to 20 times without sacrificing on area overhead and hence is suitable for high density SRAM designs in commercial applications. Finally, laser testing of SRAM based configuration memory of a Xilinx Virtex-5 FPGA is performed to analyze the behavior of SRAM based systems towards radiation strikes.

Reverse Auction Bidding is a recently developed auction method. In this form of bidding process, the roles of the bidders and the owner are interchanged in terms of the form of the economic transaction. The owner's objective is to drive the unit rates down and the bidder's objective is to maintain an acceptable profit level. A study into Reverse Auction Bidding commenced at Texas A&M University in 2004 and continues to this time, with this the eighteenth study in the series. This study is the second multi-group study in the research. In this study, a multiple group comparison was made between different numbers of bidders, with Games One, Two and Three having three, four and ten bidders respectively. All participants were faculty and students from the Department of Construction Science. The critical requirement for the participants is that they should have no prior experience using the Reverse Auction Bidding system. The eighteen studies have concentrated on new players, with future studies planned for repeat participants. A number of the recent case studies have shown personality has an impact on the performance of the bidders. However, this work was not controlled for personality, as the research objective was to determine the impact of a different number of bidders in a game. The Keirsey Temperament Sorter test was completed by all participants so that the results could be understood in terms of personality impact on the level of return to each participant. The results showed the number of bidders has a significant impact on the individual returns confirming the earlier work on varying the number of bidders. An increase in the number of bidders was shown to lead to a more competitive economic environment, which given usual economic circumstances lead to a reduction in the number of firms interested in bidding, for the self-evident economic reasons. This work points to the need to investigate a bidding group size of five or six, which is likely to be the self-constrained upper limit in a real economic system. Some interesting observations on the personality types suggest that further work is required in this area.

The purpose of the study was to provide a comprehensive framework that explains how consumer experiences within new, innovative media affect advertising effectiveness. Several concerns about previous advertising models motivated this study. For instance, advertising models traditionally have focused on message recipients’ characteristics and information processes, ignoring the significant role of media in understanding advertising effectiveness. In addition, recently developed advertising models dealing with the impact of media have been narrowly applied to a specific medium, the Internet, and have focused largely on interactivity. The proposed model and our findings highlighted the prominent roles of media novelty and presence in enhancing advertising effectiveness in an innovative, new medium that emphasizes vividness, stereoscopic 3-D. The novelty effect, created by the newness of the medium, had the power to attract viewers’ attention and the increased attention enhanced their sense of presence, the experience of being plunged into a new virtual world that advertisers constructed. The findings demonstrated that these sequential relationships can result in positive measures of advertising effectiveness, such as improved product knowledge and increased enjoyment, and ultimately more favorable attitudes toward the ad. Also our findings revealed that an irritation, such as cybersickness resulting from the stereoscopic 3-D, can hinder ad viewers’ communication processes and reduce their attention to the ad and their enjoyment of it. The model predicted that user characteristics, such as innovativeness, curiosity, and previous experience with the medium, would affect the process, but no effects were found. The current research provided advertising practitioners and researchers with opportunities to consider the significant role of media, especially innovative media, in assessing overall advertising effectiveness. For managers, it highlighted the potential of stereoscopic 3-D technology as an emerging advertising vehicle. Given the rapid changes in the media environment, it is increasingly important to understand the important roles that media play in advertising effectiveness.
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The AGC group of protein kinases (named for protein kinases A, G, and C) is found in all eukaryotes studied so far, and its members coordinate essential cellular processes including translation, metabolism, hormone response, growth, and survival. AGC kinases are intensively studied in mammals because of their connection with human diseases like cancer, diabetes, and neurological disorders. Some aspects of AGC kinase function are organism-specific, but others are conserved in highly divergent species. Several AGC kinases are regulated by the conserved 3-phosphoinositide dependent protein kinase-1 (PDK1), which is itself an AGC kinase. PDK1 regulates its substrates through phosphorylation at a conserved site in their activation loop. Here, I identify and characterize a PDK1 homologue from the moss Physcomitrella patens (PpPDK1). I show PpPDK1 phosphorylates plant AGC kinases in the activation loop, but unexpectedly lacks a lipid-binding domain, suggesting that its regulation differs from other species. In contrast to mammalian cells, PpPDK1 is not an essential gene, suggesting that AGC kinase pathways in P. patens are sufficient for survival even in the absence of activation by PpPDK1. I analyze putative PDK1 sequences from 100 different eukaryotic species, finding that many PDK1s differ from the "conventional" PDK1 found in humans. Phylogenetic analysis of these sequences suggests a complicated evolutionary history for PDK1, with the potential for unexpected functional and regulatory features. I also investigate the regulation of Adi3, an AGC kinase from tomato, through phosphorylation by PDK1. I identify a novel putative PDK1 phosphorylation site outside the kinase domain, which appears to increase Adi3 activity on a substrate. Finally, I produce a mutant version of Adi3 that can selectively utilize bulky ATP analogues. This analogue-sensitive protein may be used in a future search for direct Adi3 substrates. Together, my experiments provide insight into two members of the AGC group of protein kinases, one (PDK1) that is conserved in all eukaryotes and one (Adi3) that appears to be present only in plants. These experiments give a new perspective in our view of plant AGC kinase function and regulation.

Modeling of wind farms to determine their short circuit contribution in response to faults is a crucial part of system impact studies performed by power utilities. Short circuit calculations are necessary to determine protective relay settings, equipment ratings and to provide data for protection coordination. The plethora of different factors that influence the response of wind farms to short circuits makes short circuit modeling of wind farms an interesting, complex, and challenging task. Low voltage ride through (LVRT) requirements make it necessary for the latest generation of wind generators to be capable of providing reactive power support without disconnecting from the grid during and after voltage sags. If the wind generator must stay connected to the grid, a facility has to be provided to by-pass the high rotor current that occurs during voltage sags and prevent damage of the rotor side power electronic circuits. This is done through crowbar circuits which are of two types, namely active and passive crowbars, based on the power electronic device used in the crowbar triggering circuit. Power electronics-based converters and controls have become an integral part of wind generator systems like the Type 3 doubly fed induction generator based wind generators. The proprietary nature of the design of these power electronics makes it difficult to obtain the necessary information from the manufacturer to model them accurately. Also, the use of power electronic controllers has led to phenomena such as sub-synchronous control interactions (SSCI) in series compensated Type 3 wind farms which are characterized by non-fundamental frequency oscillations. SSCI affects fault current magnitude significantly and is a crucial factor that cannot be ignored while modeling series compensated Type 3 wind farms. These factors have led to disagreement and inconsistencies about which techniques are appropriate for short circuit modeling of wind farms. Fundamental frequency models like voltage behind transient reactance model are incapable of representing the majority of critical wind generator fault characteristics such as sub-synchronous interactions. The Detailed time domain models, though accurate, demand high levels of computation and modeling expertise. Voltage dependent current source modeling based on look up tables are not stand-alone models and provide only a black-box type of solution. The short circuit modeling methodology developed in this research work for representing a series compensated Type 3 wind farm is based on the generalized averaging theory, where the system variables are represented as time varying Fourier coefficients known as dynamic phasors. The modeling technique is also known as dynamic phasor modeling. The Type 3 wind generator has become the most popular type of wind generator, making it an ideal candidate for such a modeling method to be developed. The dynamic phasor model provides a generic model and achieves a middle ground between the conventional electromechanical models and the cumbersome electromagnetic time domain models. The essence of this scheme to model a periodically driven system, such as power converter circuits, is to retain only particular Fourier coefficients based on the behavior of interest of the system under study making it computationally efficient and inclusive of the required frequency components, even if non-fundamental in nature. The capability to model non-fundamental frequency components is critical for representing sub-synchronous interactions. A 450 MW Type 3 wind farm consisting of 150 generator units was modeled using the proposed approach. The method is shown to be highly accurate for representing faults at the point of interconnection of the wind farm to the grid for balanced and unbalanced faults as well as for non-fundamental frequency components present in fault currents during sub-synchronous interactions. Further, the model is shown to be accurate also for different degrees of transmission line compensation and different transformer configurations used in the test system.

Humans impact rivers in many ways that modify ecological processes yielding ecosystem services. In order to mitigate anthropogenic impacts, scientists are challenged to understand interactions among physicochemical factors affecting large river food webs. An understanding of socioeconomic factors also is critical for ecosystem management. In this dissertation, I explore spatiotemporal patterns in floodplain river food webs and political barriers to management of environmental flows, an important factor influencing river ecology. In Chapter II, I reviewed the scientific literature to test conceptual models of river food webs and predictions of environmental factors that might produce variation in basal production sources supporting consumer biomass. My review indicates that algae are the predominant production source for large rivers worldwide, but consumers assimilate C3 plants in rivers 1) with high sediment loads and low transparency during high flow pulses, 2) with high dissolved organic matter concentrations, and 3) following periods of high discharge or leaf litter fall that increase the amount of terrestrial material in the particulate organic matter pool. In Chapter III, I descrobe field research conducted to examine relationships among hydrology, nutrient concentrations, turbidity, and algal primary production and biomass in the littoral zone of five rivers in Texas, Peru, and Venezuela differing in physicochemical conditions. I used stable isotope signatures to estimate contributions of algal-versus terrestrial-based production sources to consumers during different hydrologic periods. My research indicates that during flow pulses in floodplain rivers, a decrease in algal biomass and productivity, combined with increased inputs of terrestrial organic matter, can result in increased terrestrial support of metazoan consumers in the aquatic food web. In 2007, Texas Senate Bill 3 directed that environmental flow recommendations be developed for river basins. Despite emphasis on use of the "best available science" to develop environmental flow regimes and "stakeholder involvement" to address needs of all water users, for the first two basins to complete the SB3 process, final environmental flow rules did not mimic a natural flow regime. In Chapter IV, I reviewed this process, concluding that incentives for river authorities to increase compromise with diverse stakeholders should result in more sustainable management of freshwater.

This dissertation reports the development of a new multi-band multi-output synthesizer, 1/2 dual-injection locked divider, 1/3 injection-locked divider with phase-tuning, and 1/3 injection-locked divider with self-injection using 0.18-micrometer CMOS technology. The synthesizer is used for a multi-band multi-polarization radar system operating in the K- and Ka-band. The synthesizer is a fully integrated concurrent tri-band, tri-output phase-locked loop (PLL) with divide-by-3 injection locked frequency divider (ILFD). A new locking mechanism for the ILFD based on the gain control of the feedback amplifier is utilized to enable tunable and enhanced locking range which facilitates the attainment of stable locking states. The PLL has three concurrent multiband outputs: 3.47-4.313 GHz, 6.94-8.626 GHz and 19.44-21.42-GHz. High second-order harmonic suppression of 62.2 dBc is achieved without using a filter through optimization of the balance between the differential outputs. The proposed technique enables the use of an integer-N architecture for multi-band and microwave systems, while maintaining the benefit of the integer-N architecture; an optimal performance in area and power consumption. The 1/2 dual-ILFD with wide locking range and low-power consumption is analyzed and designed together with a divide-by-2 current mode logic (CML) divider. The 1/2 dual-ILFD enhances the locking range with low-power consumption through optimized load quality factor (QL) and output current amplitude (iOSC) simultaneously. The 1/2 dual-ILFD achieves a locking range of 692 MHz between 7.512 and 8.204 GHz. The new 1/2 dual-ILFD is especially attractive for microwave phase-locked loops and frequency synthesizers requiring low power and wide locking range. The 3.5-GHz divide-by-3 (1/3) ILFD consists of an internal 10.5-GHz Voltage Controlled Oscillator (VCO) functioning as an injection source, 1/3 ILFD core, and output inverter buffer. A phase tuner implemented on an asymmetric inductor is proposed to increase the locking range. The other divide-by-3 ILFD utilizes self-injection technique. The self-injection technique substantially enhances the locking range and phase noise, and reduces the minimum power of the injection signal needed for the 1/3 ILFD. The locking range is increased by 47.8 % and the phase noise is reduced by 14.77 dBc/Hz at 1-MHz offset.

The biological function of RNA is often linked to an ability to adopt one or more mutually exclusive conformational states or isomers, a characteristic that distinguishes this biomolecule from proteins. Two examples of conformationally inconverting RNAs were structurally investigated. The first is found in the 3' untranslated region (UTR) of the coronavirus mouse hepatitis virus (MHV). A proposed molecular switch between mutually exclusive stable stem loop and pseudoknot conformations was investigated using thermal unfolding methods, NMR spectroscopy, sedimentation velocity ultracentrifugation and fluorescence resonance energy transfer (FRET) spectroscopy. Utilizing a "divide and conquer" approach we establish that the independent subdomains are folded as predicted by the proposed model and that a pseudoknotted conformation is accessible. Using the subdomains as spectral markers for the investigation of the intact 3' UTR RNA, we show that the 3' UTR is indeed a superposition of a double stem conformation and a pseudoknotted conformation in the presence of KCl and MgCl2. In the absence of added salt however, the 3' UTR adopts exclusively the double stem conformation. Analysis of the pseudoknotted stem reveals only a marginally stable folded state (deltaG25 = 0.5 kcal mol-1, tm = 31 oC) which makes it likely that a viral or host encoded protein(s) is required to stabilize the pseudoknotted conformation. A second conformationally interconverting RNA system investigated is an RNA element that stimulates -1 programmed ribosomal frameshifting in the human Ma3 gene. Structural analysis of the frameshifting element reveals a dynamic equilibrium between a functionally inactive double stem loop conformation and the active pseudoknotted conformation. Thermal melting and NMR spectroscopy reveal that the double stem loop is the predominant conformation in the absence of added KCl or MgCl2. The addition of KCl and MgCl2 results in the formation of a pseudoknot conformation. This conformation is dominant in solution only when the competing double stem loop conformation is abrogated by mutation. Functional studies of the Ma3 pseudoknot reveal that abrogation of double stem conformation increases frameshift stimulation by 2-fold and indicates that the pseudoknot is the active conformation.

Modern engine technologies are subject to increasingly tighter emission standards and recent number-based regulations have become a new challenge, since historically only a mass-based regulation needed to be met. This evolution derives from the need to control the emissions of very fine particles, that are believed to cause more damage than larger ones. The aim of the present work is to provide further guidance in understanding the mechanisms of particle emission processes in Spark-Ignition (SI) engines. By means of both numerical and experimental investigations, it tries to answer some still open questions related to this complex topic. Different fuels are considered, such as gasoline and other promising cleaner alternatives for the future, including natural gas. 3-D Computational Fluid Dynamics simulation are used as useful additional tool to investigate the fuel-related soot emissions and help explain the experimental-derived results. The modified version of the KIVA-3V code, developed at the Engine Research Center (ERC) of the University of Wisconsin-Madison, is used for the present modeling work. It includes improvements in its ignition, combustion and emission models. In particular, a semi-detailed soot model and a chemical kinetic model, including Poly-Aromatic Hydrocarbon formation, are coupled with a SI model and the G equation flame propagation model for the engine simulations and for predictions of soot mass and particulate number density. The present work improves and extends the laminar flame speed correlations for several fuels of practical use in order to assure the correct prediction of combustion phasing and in-cylinder pressure evolution. The effects of a load increase achieved by pure oxygen addition in gasoline SI engines, as well as, the influence of natural gas composition on combustion are investigated. Furthermore, additional extensive experimental investigations provide more insights about the effects of lubricant oil on particle emissions from both gasoline and natural gas SI engines. In this last case both Port Fuel and Direct Injection mode are considered. The experimental tests were performed at the �Istituto Motori CNR�, Italy.

Morphological transitions in Wangiella dermatitidis, a causative agent of human phaeohyphomycosis, influence virulence processes in this polymorphic fungus. My project first involved the cloning and characterizion of the β(1→3) glucan synthase gene WdFKS1, which encodes the enzyme's catalytic subunit, followed by cloning and characterizing the WdRHO1 gene, which encodes its regulatory subunit. To better understand the Rho-type GTPase-mediated regulation of cell polarity and its role in fungal morphological transitions, a homologue of WdRAC1 from a W. dermatitidis was subsequently identified by degenerate PCR and gene walking. Gene deletions of WdFKS1 and WdRHO1 in haploid W. dermatitidis were lethal, whereas the deletion of WdRAC1 was not. RNA interference on WdFKS1 mRNA expression resulted in incomplete septa and damaged cell wall integrity, as well as slow growth rate in W. dermatitidis. Overexpression studies, after site-specific integrations of WdRHO1 and WdRAC1 alleles under control of the glaA promoter into the nonessential WdPKS1 locus, showed the different alleles had different effects on the cell morphological development. For example, whereas overexpression of the wdrho1⁺ allele did not affect the growth rate of W. dermatitidis, the overexpression of wdrho1[superscript G14V], a constitutively active mutation, slowed growth and repressed true filamentous hyphal growth by promoting pseudohyphal growth. While the deletion of WdRAC1 did not affect growth, its loss retarded polarized hyphal growth in a hyphal-inducing minimal medium. Moreover, three new phenotypes of a previously derived WdCDC42 deletion mutant were discovered during this study: in the first, the wdcdc42[Delta] mutant displayed cell lysis when incubated in YPMaltose medium at 37°C; in the second, a dark budding neck abnormality was found after Calcoflour staining; and in the third, the wdcdc42[Delta] mutant displayed no branching during true hyphal growth. Interestingly, the overexpression of wdrac1[superscript G16V] complemented the second and the third phenotypes caused by the WdCDC42 deletion. In addition, the wdcdc42[Delta]/wdrac1[superscript G16V] double mutant unexpectedly displayed an interrupted carotenogenesis pathway. These results support that in W. dermatitidis, Rho-type GTPases play essential roles in growth rate determination and cellular morphogenesis, especially while producing polarized hyphal growth during its many morphological transitions.
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The Pezizomycotina, commonly known as the filamentous fungi, are a diverse group of organisms that have a major impact on human life. The filamentous fungi diverged from a common ancestor approximately 200 – 700 million years ago. Because of the diversity and the wealth of biological and genomic tools for the filamentous fungi it is possible to track the evolutionary history of genes and gene networks in these organisms. In this dissertation I focus on the evolution of two genes (lolC and lolD) in the LOL secondary metabolite gene cluster in Epichloë and Neotyphodium genera, the evolution of the MAP kinase-signaling cascade in the filamentous fungi, the regulation of the gene networks involved in asexual development in Neurospora crassa, and the identification of two genes in the N. crassa asexual development gene network, acon-2 and acon-3. I find that lolC and lolD originated as an ancient duplication in the ancestor of the filamentous fungi, which were later recruited in the LOL gene cluster in the fungal endophyte lineage. In the MAP kinase-signaling cascade, I find that the MAPK component is the most central gene in the gene network. I also find that the MAPK signaling cascade originated as three copies in the ancestor to eukaryotes, an arrangement that is maintained in filamentous fungi. My observations of gene expression profiling during N. crassa asexual development show tissue specific expression of genes. Both the vegetative mycelium and the aerial hyphae contribute to the formation of macroconidiophores. Also, with the help of genomic tools recently developed by researchers in the filamentous fungal community, I identified NCU00478 and NCU07617 as the genes with mutations responsible for two aconidial strains of N. crassa, acon-2 and acon-3 respectively.

Coaches in Canada are trained through the National Coaching Certification Program (NCCP) administered by The Coaching Association of Canada. The NCCP is the national standard and recently shifted its educational emphasis from the transfer of knowledge to the development of coaching competency. As a result, coaches are required to demonstrate competency in a specified sport and level prior to being awarded a coaching certification. The purpose of this study was to establish validity and reliability of an NCCP coaching competency assessment. Gymnastics Canada agreed to have their Community Sport coaching competency instrument tested for content validity, face validity, inter-rater reliability and intra-rater reliability. Their original assessment instrument was revised during content validity testing based on recommendations from five coaching context experts. Removing six items, adding one item, and revising four items for clarity modified the original 48-item instrument. An assessment of overall competency was added to the instrument to rate the coach as Beginner, Competent, Proficient, or Expert. Three certified coaches confirmed the face validity of the modified instrument. Reliability tests were conducted on the ratings provided by ten experts who observed a coach’s lesson on video. The result was a moderate level of inter-rater reliability, displayed by an Agreement Coefficient (AC1) of 0.43 and a Percent of Agreement (PA) of 72%. Nine of the ten raters assessed the coach’s performance as Competent, Proficient or Expert, while one of the ten rated the coach as Beginner. After a repeat observation of the same coaching performance, the intra-rater reliability of five raters resulted in agreement levels of Moderate (AC1 = 0.45; PA = 67%), Substantial (AC1 = 0.75, 0.77; PA = 81%, 89%), and Almost Perfect (AC1 = 0.87, 0.82; PA = 93%, 90%). The modified instrument has content and face validity. However, its usefulness is impacted by variability in rater preferences. The inter-rater reliability results attest to concerns about the consistency in assessment of a coaching certificate for Community Sport Artistic Gymnastics coaches. Suggestions to improve the reliability of the instrument include training raters to classify the coach’s competency on a standardized scale of sport-specific expertise.

Previous research has shown how home and parental characteristics support or hinder the development of children’s self-regulation in the family context. There have only been limited attempts to understand these mechanisms in early childhood education settings. This study used the NICHD Study of Early Child Care (when participating children were 36-months old) to examine the relations among various aspects of the early childhood education setting, the interactions in the setting, and children’s self-regulation in center-based and home-based settings. Structural equation modeling was used to test a model proposing the deconstruction of early childhood education quality into structural (i.e., environmental and caregiver characteristics) and process quality components (i.e., positive and negative interactions) and to examine these as predictors of three-years old children’s self-regulation abilities. A meditational model was tested in which positive and negative interactions in the classroom mediated the relations between the structural characteristics and self-regulation. There were three important findings. First, although there were no consistent patterns of associations between structural features and self-regulation across the two types of care, there were more significant relationships in home-based care compared to center-based care. These findings showed that the home-based caregiver characteristics were more closely tied to the processes in the classroom than those characteristics of caregivers in center care. Second, both positive and negative caregiving were associated with children’s compliance, which suggested that compliance may have been influenced differently by process quality compared to other self-regulation measures, such as self-control and emotion-, behavior-, and attention-regulation. It may be that high rates of compliance may be markers of highly restrictive caregiving rather than the result of good quality caregiving. Third, there were very few significant relationships between process quality measures and children’s self-regulation measures, which suggested that commonly used process quality measures may not be capturing the processes that are most important for the development of self-regulation.
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It is impossible to remove methylmercury (MeHg) from biological systems because MeHg is found throughout our environment in many fresh and salt water fish. The consumption of fish is important to human nutrition and health. The mechanism of MeHg neurotoxicity must be understood to minimize adverse exposure consequences. The dissertation objective was to: 1) compare mechanisms of MeHg neurotoxicity between animals exposed as adults and those exposed during gestation, and 2) develop an in vitro test model of in vivo MeHg exposure. Total mercury (Hg) levels in tissue / cells were determined by combustion / trapping / atomic absorption. Cell death was determined by Fluoro-Jade histochemical staining and activated caspase 3 immunohistochemistry for in vivo studies, and Trypan blue exclusion, lactate dehydrogenase activity, and cytotoxicity assays for in vitro studies. Mitochondrial membrane potential (MMP), intracellular calcium ion concentration ([Ca2+]i), and production of reactive oxygen species (ROS) were determined using fluorescence microscopy or microplate reader assays. Young adult C57Bl/6 mice were exposed to a total dose of 0, 1.0, or 5.0 mg/kg body weight MeHg divided over postnatal days (P)35 to 39. Pregnant female mice were exposed to a total does of 0, 0.1, or 1.0 mg/kg body weight MeHg divided over gestational days (G)8 to 18. SY5Y cells were exposed to 0, 0.01, 0.1, or 1.0 ?M MeHg or HgCl2 for 24, 48, or 72 hours. Total Hg in brains of young adult mice, mouse pups, and SY5Y cells accumulated in a dose-dependent manner. Cell death increased in SY5Y cells exposed to the highest concentrations of MeHg and HgCl2 used in this study. Cell death increased in the molecular and granule cerebellar cell layers of young adult mice exposed to the highest doses of MeHg used in this study. P0 mouse pups showed no increase in cell death within the cerebellum following MeHg exposure. Cerebella of mice at P10 exhibited decreased dying cells only in the external germinal layer. Low concentrations of MeHg affected MMP in both in vivo and in vitro studies, but did not result in decreased MMP typically associated with higher MeHg concentrations. [Ca2+]i was increased throughout the in vivo experiments in an age- , sexand brain region-dependent manner. Generation of ROS was decreased in both in vivo and in vitro studies with both the MeHg and HgCl2 (in vitro) treatments. In summary, low and moderate MeHg exposure, both in vivo and in vitro, altered mitochondrial function, Ca2+ homeostasis, and ROS differently than what is reported in the literature for higher MeHg exposure concentrations. SY5Y cells were sensitive to low-levels of MeHg and HgCl2 and responded similarly to cells in the whole animal studies, thus making SY5Y cells realistic candidates for mechanistic MeHg studies. Cell culture and whole animal neuronal functional studies at chronic low-level MeHg exposure are limited. These data suggest that low-levels of MeHg may affect neuronal function. Therefore, further chronic low-level MeHg neuronal functional studies are warranted.

There has recently been growing interest in using physical layer network coding techniques to facilitate information transfer in wireless relay networks. The physical layer network coding technique takes advantage of the additive nature of wireless signals by allowing two terminals to transmit simultaneously to the relay node. This technique has several performance benefits, such as improving utilization and throughput of wireless channels and reducing delay. In this thesis, we present an algorithm for joint decoding of two unsynchronized transmitters to a modulo-2 sum of their transmitted messages. We address the problems that arise when the boundaries of the signals do not align with each other and when their phases are not identical. Our approach uses a state-based Viterbi decoding scheme that takes into account the timing offsets between the interfering signals. As a future research plan, we plan to utilize software-defined radios (SDRs) as a testbed to show the practicality of our approach and to verify its performance. Our simulation studies show that the decoder performs well with the only degrading factor being the noise level in the channel.

Il presente documento intende studiare in maniera approfondita tecniche e proporre metodologie atte a ricucire la “dicotomia” tra forma architettonica e performance strutturale con l’ausilio e il riconoscimento dell’architettura di San Paolo (o Scuola Paulista – o ancora Scuola Paulista Brutalista), esaminando in maniera critica il periodo tra il 1945 e il 1970, attraverso due sezioni differenti e non immediatamente complementari. Tale difficoltà è rappresentata dalla differenza delle tematiche trattate nei successivi capitoli, all’interno dei quali ci si vuole focalizzare sui concetti pragmatici che conducono alla creazione di determinate soluzioni architettoniche, a loro volta guidate da rappresentazioni “ideali” fondamentali dell’architettura della Scuola di São Paulo. Il documento è suddiviso in due differenti macro-sezioni che consentono al lettore di essere guidato nella ricerca condotta, partendo dalle motivazioni per il quale determinate architetture si sono prestate più di altre nel fungere come “background” della ricerca, approfondendo allo stesso tempo eventi storici, punti salienti, caratteri distintivi e pratici nella definizione dell’architettura paulista. La prima sezione si propone di fornire alcune indagini bibliografiche, con approccio il più possibile critico. Al contempo, vengono raccolti riferimenti sparsi e sconnessi sull'Architettura Paulista, permettendo di indagare principalmente sull’incidenza della tematica forma-struttura in riferimento a specifiche opere esistenti e/o esistite nel panorama della Escola Paulista, le quali sono in grado di riflettere il valore intrinseco del linguaggio architettonico accostato con la stabilità e l’innovazione strutturale. In particolare, viene concentrata l’attenzione su specifiche opere dei maestri Vilanova Artigas e Paulo Mendes da Rocha – pionieri e fondatori della Scuola di San Paolo, il cui studio ha permesso di riconoscere le caratteristiche estetiche ed etiche dell'architettura Brutalista Paulista, sottolineando la fusione di caratteri morfologici e strutturali che caratterizzano tali opere. La seconda sezione esamina sistematicamente le caratteristiche dell’Architettura Paulista e, in special modo, di specifici casi di studio caratterizzati dall’ampia campata. Vengono così introdotte specifiche metodologie di Ottimizzazione Strutturale (SO) utili nella progettazione formale delle strutture - o parte di esse. Le metodologie studiate nel campo della SO offrono la possibilità al progettista di poter controllare (e dunque decidere) la forma finale di organismi architettonici e delle sue componenti, avendo allo stesso tempo la certezza di ottenere strutture performanti. I criteri di ottimizzazione proposti e sperimentati - riguardanti maggiormente casi di trave a sezione variabile - saranno applicati sui casi di studio scelti, dimostrando le potenzialità degli algoritmi sviluppati, i quali fungono da punto di connessione tra la progettazione della forma architettonica e quella della performance strutturale. La documentazione sviluppata – grazie anche agli approfondimenti condotti sui casi di studio – ha consentito di introdurre le prime osservazioni sull’argomento riguardante l’Ottimizzazione Strutturale e sull’importanza che riveste in ambito architettonico, costruttivo e sulla tematica del rapporto forma-struttura. Alla presentazione di metodologie tradizionali di SO segue l’introduzione di metodologie sperimentali applicate sull’elemento strutturale trave - oggetto di sperimentazioni - riportando le principali teorie che la caratterizzano. La valutazione della precisione dei metodi proposti è oggetto di studio nel quale, oltre a constatare l’errore effettivo commesso nell’utilizzo dei metodi riportati dalla letteratura classica sull’argomento, verrà effettuato un primo studio di SO nell’ambito di uno o più casi studio esistenti estrapolati dalle architetture della Scuola Paulista. L’attenzione è dunque focalizzata verso quei criteri di ottimizzazione strutturale che hanno le potenzialità di fungere da elemento di contatto tra una struttura correttamente realizzata e l’equivalente formale che ne deriva, procedendo tramite l’applicazione di criteri sperimentali di ottimizzazione volti alla determinazione di sezioni ottimali valutando differenti combinazioni di carico. Si tratterà dunque, nel corso della presente dissertazione, la determinazione di sviluppo ottimale di travi secondo la trattazione proposta da Timoshenko (il quale fa proprie le teorie di Eulero – Bernoulli, utilizzando la componente di sforzo data dal momento flettente agente per la determinazione della condizione di ottimo per lo sviluppo longitudinale della trave) e di studio e sviluppo di elementi strutturali dalla funzione autoportante (Modelli Comparativi - Appendice). In dettaglio, Nel Capitolo I saranno esaminati i caratteri fondanti della Escola Paulista nel tentativo introdurre le prime considerazioni sulla tematica forma-struttura. La finalità di tali analisi consiste nel voler comprendere la rilevanza del rapporto tra caratteri formali e strutturali dell’architettura in ambito linguistico e costruttivo, fissando le linee guida per la lettura dell’organismo architettonico attraverso la sua decostruzione nelle varie discipline/ragioni che ne concorrono alla formazione. Il Capitolo II descrive criticamente, attraverso il ridisegno e la rielaborazione della documentazione consultata, i casi di studio considerati nella ricerca, anche in questo caso rimarcando il ruolo che gli elementi strutturali hanno rappresentato nella creazione dell’organismo architettonico e di come nello stesso elemento strutturale sia racchiuso il linguaggio dell’architettura e la volontà di denunciare specifiche ideologie. La seconda parte del documento si sviluppa in due capitoli. Essi rappresentano il fulcro della ricerca in quanto vengono presentati i principali criteri sperimentali di ottimizzazione strutturale testati su travi a sezione variabile con geometrie vincolate attraverso l’applicazione di tecniche computazionali (geometria computazionale) accostate all’uso di algoritmi genetici evolutivi per la risoluzione del problema (Capitolo III). Le metodologie proposte verranno valutate attraverso la loro applicazione nel Capitolo IV, all’interno del quale viene discussa la validità dei processi e l’utilità in fase di concepimento di un’opera architettonica, sottolineando l’importanza e l’impatto che gli algoritmi presentati potrebbero avere nella definizione di un nuovo metodo progettuale che possa guidare simultaneamente la progettazione architettonica e quella strutturale. I risultati ottenuti nel capitolo finale e argomentati nella sezione dedicata alle conclusioni, non hanno la prepotenza di contestare la forma delle strutture esistenti, ma vogliono essere testimonianza di come sia possibile avvicinare due mondi, quelli dell’ingegneria e dell’architettura, che hanno subito una scissione nel tempo, ma che restano profondamente connessi. Inoltre, le indagini condotte nella dissertazione desiderano ardentemente sollevare alcuni argomenti iniziali, piuttosto che arrivare a conclusioni definitive, ancora ampiamente lontane data l’estrema complessità di opere che non sono ancora del tutto state raccontate, svelate e demistificate dal punto di vista pratico dell’architettura.
This document intends to study in-depth techniques and propose methodologies to mend the "dichotomy" between architectural shape and structural performance with the help and recognition of the architecture of São Paulo (or Paulist School - or Paulist Brutalist School), by critically examining the period between 1945 and 1970, through two different and not immediately complementary sections. This difficulty is represented by the difference in the themes dealt with in the following chapters, within which we want to focus on the pragmatic concepts that lead to the creation of specific architectural solutions, in turn, guided by fundamental "ideal" representations of the architecture of the School of São Paulo. The document is divided into two different macro-sections, allowing the reader to be guided in the research conducted. Starting from the reasons why specific architectures have lent themselves more than others in representing the "background" of the investigation, the dissertation also deepens historical events, salient points, distinctive and practical characters in the definition of Paulist architecture. The first section aims to provide bibliographic investigations with an approach that is as critical as possible. At the same time, scattered and disconnected references on Paulista Architecture are collected. This allows investigating mainly the incidence of the shape-structure theme concerning specific current and existing works in the panorama of the Escola Paulista, able to reflect the intrinsic value of the architectural language combined with stability and structural innovation. Attention is focused on specific works by the masters Vilanova Artigas and Paulo Mendes da Rocha - pioneers and founders of the School of São Paulo, whose study has allowed us to recognize the aesthetic and ethical characteristics of Paulist Brutalist architecture, underlining the fusion of morphological and structural characters that characterize these works. The second section systematically examines the characteristics of Paulista Architecture and of specific case studies characterized by wide spans. Specific Structural Optimization (OS) methodologies are thus introduced, useful in the shape-design of structures - or part of them. The methods developed in the OS field offer the possibility for the designer to control (and therefore decide) the final shape of architectural organisms and their components while having the certainty of obtaining performing structures. The proposed and tested optimization criteria - mainly concerning cases of variable section beams - will be applied to the selected case studies, demonstrating the developed algorithms' potential, representing the linking point between the design of the architectural shape and that of the structural performance. Thanks to the in-depth analysis conducted on the case studies, the developed documentation made possible the introduction to the first observations on the topic concerning Structural Optimization and its importance in architectural design, construction, and the theme of the shape-structure relationship. The presentation of traditional SO methodologies is followed by introducing experimental methods applied to the structural beam element - subject to experimentation - reporting the main theories that characterize it. The evaluation of the precision of the proposed methods is the subject of study. In fact, in addition to ascertaining the actual error committed to using the methods reported in the classical literature on the subject, one first study of OS will be carried out in the context of one or more case studies extrapolated from the architecture of the Paulista School. Attention is therefore focused on those structural optimization criteria that potentially represent a contact element between a correctly constructed structure and the resulting shape-equivalent, proceeding through the application of experimental optimization criteria to determine optimal sections by evaluating different load combinations. Therefore, in this dissertation, we will determine the optimal development of beams according to the treatment proposed by Timoshenko and the development of structural elements with a self-supporting function (Comparative Models - Appendix). In detail, in Chapter 1, the founding characteristics of the Escola Paulista, to introduce the first considerations on the form-structure theme, will be examined. The purpose of these analyses is to understand the relevance of the relationship between shape features and structural architecture characteristics in the linguistic and constructive field, setting the guidelines for reading the architectural organism through its deconstruction in the various disciplines/reasons that concur to its creation. Through the redraws and elaboration of the consulted documentation, Chapter 2 critically describes the case studies considered in the research, underlining the role of specific structural elements played in the creation of the architectural organism and how in the structural parts itself, the architectural language, and the will to denounce specific ideologies, are contained. The second part of the document is developed in two chapters. They represent the core of the research as the main experimental criteria of structural optimization tested on variable section beams with constrained geometries are presented through the application of computational techniques (computational geometry) combined with the use of evolutionary genetic algorithms for solving the problem. (Chapter III). The proposed methodologies will be evaluated through their application in Chapter IV. First, the validity of the processes and the usefulness in the conception phase of architectural work are discussed. Likewise, the impact of the algorithms presented in defining a new design method that can guide architectural and structural design simultaneously is debated. The results obtained in the final chapter and then discussed in the section dedicated to the conclusions do not have the pride to challenge the architectural shape of the case studies. Still, they want to testify how it is possible to bring two worlds, engineering, and architecture, which have undergone a split over time but remain deeply connected. Furthermore, the conducted investigations are eager to raise some initial arguments rather than reach definitive conclusions, still widely distant given the extreme complexity of works that have not yet been fully narrated, revealed and demystified from the practical architectural point of view.