Does ragging play a role in medical student depression - Cause or effect?

Background: Medical students experience a lot of stress what may contribute to symptoms of depression. In this study we set out to look at the environmental factors which may be contributing in one medical school in Brazil. Methods: We assessed depressive symptoms using Beck's Depression Inventory in 465 and 267 medical students in 2001 and 2006 respectively. We explored possible social and environmental causes using qualitative data. Results: Nearly 15% scored above the cut off for depression in both the samples. Males in the pre-clinical stage in 2006 showed an increase in depressive symptoms than males in the same cycle in 2001 (aOR = 7.36 [95% CI = 0.85-63.5] p = 0.07). Qualitative data confirmed that factors such as ragging and low social involvement were correlated with depressive symptoms in pre-clinical stage males. Limitations: The sample size was small both for quantitative and qualitative aspects of the study. Conclusions: It appears that ragging plays an important role in the genesis of depressive symptoms in medical students. (C) 2012 Elsevier B.V. All rights reserved.

Motor Neuron Disease and Acquired Axonal Neuropathy Association in HIV Infection: Case Report and Update

Background: A possible viral etiology has been documented in the genesis of motor neuron disorders and acquired peripheral neuropathies, mainly due to the vulnerability of peripheral nerves and the anterior horn to certain viruses. In recent years, several reports show association of HIV infection with Amyotrophic Lateral Sclerosis Syndrome, Motor Neuron Diseases and peripheral neuropathies. Objective: To report a case of an association between Motor Neuron Disease and Acquired Axonal neuropathy in HIV infection, and describe the findings of neurological examination, cerebrospinal fluid, neuroimaging and electrophysiology. Methods: The patient underwent neurological examination. General medical examinations were performed, including, specific neuromuscular tests, analysis of cerebrospinal fluid, muscle biopsy and imaging studies. Results and Discussion: The initial clinical presentation of our case was marked by cramps and fasciculations with posterior distal paresis and atrophy in the left arm. We found electromyography tracings with deficits in the anterior horn of the spinal cord and peripheral nerves. Dysphagia and release of primitive reflexes were also identified. At the same time, the patient was informed to be HIV positive with high viral load. He received antiretroviral therapy, with load control but with no clinical remission. Conclusion: Motor Neuron disorders and peripheral neuropathy may occur in association with HIV infection. However, a causal relationship remains uncertain. It is noteworthy that the antiretroviral regimen may be implicated in some cases.

Estudo cinético de um episódio agudo de translocação bacteriana e de suas repercussões imunológicas e microcirculatórias em ratos

Introdução: Muitos pacientes ainda morrem devido a infecções. Crescentes relatos da literatura atual têm atribuído ao intestino o papel de agravamento de doenças graves, e/ou a sua participação na gênese da sepse por mecanismo de translocação bacteriana (TB). Objetivo: Avaliar de forma cinética a TB e suas repercussões microcirculatórias e imunológicas. Método: Ratos Wistar-EPM (n=162) foram aleatoriamente distribuídos em grupo Sham (n=72) e grupo TB (n=90), e avaliados nos períodos de 2h, 6h, 24h, 72h, 7 e 14 dias, em relação a índice de translocação bacteriana; microscopia intravital; perfusão tecidual; e componentes celulares e humorais da linfa mesentérica por linfograma, citometria de fluxo e CBA-Flex. Resultado: A TB ocorreu somente no grupo TB e foi expressiva nas primeiras 24 horas tornando-se negativa somente com 7 dias. A conseqüência de um episódio de TB repercutiu na celularidade e citocinas pró e antiinflamatórias da linfa mesentérica associado a lesões da microcirculação e hipoperfusão tecidual de forma local e sistêmica. A citometria de fluxo mostrou que a linfa mesentérica eferente pós TB difere significativamente do grupo Sham quanto a número e subpopulação de linfócitos. Conclusão: Um episódio agudo de TB determinou uma recuperação máxima bacteriana com 6 horas e sua completa depuração entre 3 e 7 dias, além de provocar alterações da microcirculação intestinal e sistêmica associadas à ativação do GALT, principalmente no período de permanência das bactérias translocadas no hospedeiro, sendo a via linfática mesenterial uma importante rota na intercomunicação imunológica entre o ambiente intestinal e sistêmico.

Efeitos da Prostaglandina E1 na gênese de capilares sanguíneos em músculo esquelético isquêmico de ratos: estudo histológico e ultra-estrutural

Objetivo: o objetivo deste trabalho foi estudar os aspectos morfológicos e ultra-estruturais na gênese de capilares sanguíneos em músculo esquelético do membro caudal de ratos submetidos à isquemia sob a ação da Prostaglandina E1 (PGE1), administrada por via intramuscular ou endovenosa. Métodos: foram utilizados 60 ratos (Rattus norvegicus albinus), linhagem Wistar-UEM, distribuídos aleatoriamente em três grupos de 20, redistribuídos igualmente em dois subgrupos, observados no 7o e 14o dias, sendo um grupo controle onde apenas foi provocada a isquemia no membro, outro com a isquemia e a injeção da PGE1 via intramuscular (IM), e outro com a isquemia e a injeção da PGE1 endovenosa (EV). Para análise dos resultados, foram realizadas a coloração com hematoxilina & eosina (HE), a imuno-histoquímica e a microscopia eletrônica de transmissão (MET). Resultados: constatou-se um aumento estatisticamente significante no número de capilares nos subgrupos com o uso da PGE1 IM e EV, através da contagem nos cortes corados com HE. Houve marcação de capilares e vasos de maior calibre nestes mesmos subgrupos, porém, esta reação não foi eficiente para a quantificação dos capilares. Na MET encontraram-se evidências de formação de novos capilares. Conclusões: a PGE1, administrada por via IM ou EV, promoveu, após 14 dias de observação, um aumento no número de capilares no músculo esquelético de ratos submetido à isquemia, identificáveis histologicamente com a coloração em HE. Na análise ultra-estrutural encontraram-se alterações que sugerem, nos animais sob a ação da PGE1, que a neoformação vascular possa ter ocorrido por angiogênese e vasculogênese. A imuno-coloração, apesar da marcação de capilares e vasos maiores, não permitiu estabelecer uma correlação com o aumento de vasos encontrados na coloração com HE.

Indagine sulla nozione di fatto urbano ne L'architettura della città di Aldo Rossi

Questa tesi di dottorato di ricerca ha come oggetto la nozione di fatto urbano elaborata e presentata da Aldo Rossi nel libro L’architettura della città edito nel 1966. Ne L’architettura della città sono molteplici le definizioni e le forme con cui è enunciata la nozione di fatto urbano. Nel corso della tesi si è indagato come la costruzione nel tempo di questo concetto è stata preceduta da diversi studi giovanili intrapresi dal 1953, poi riorganizzati e sintetizzati a partire dal 1963 in un quaderno manoscritto dal titolo “Manuale di urbanistica”, in diversi appunti e in due quaderni manoscritti. Il lavoro di ricerca ha ricostruito la formulazione della nozione di fatto urbano attraverso gli scritti di Rossi. In questa direzione la rilevazione della partecipazione di Rossi a dibattiti, seminari, riviste, corsi universitari o ricerche accademiche è apparsa di fondamentale importanza, per comprendere la complessità di un lavoro non riconducibile a dei concetti disciplinari, ma alla formazione di una teoria trasmissibile. Il tentativo di comprendere e spiegare la nozione di fatto urbano ha condotto ad esaminare l’accezione con cui Rossi compone L’architettura della città, che egli stesso assimila ad un trattato. L’analisi ha identificato come la composizione del libro non è direttamente riferibile ad un uso classico della stesura editoriale del trattato, la quale ha tra i riferimenti più noti nel passato la promozione di una pratica corretta come nel caso vitruviano o un’impalcatura instauratrice di una nuova categoria come nel caso dell’Alberti. La mancanza di un sistema globale e prescrittivo a differenza dei due libri fondativi e il rimando non immediato alla stesura di un trattato classico è evidente ne L’architettura della città. Tuttavia la possibilità di condurre la ricerca su una serie di documenti inediti ha permesso di rilevare come negli scritti a partire dal 1953, sia maturata una trattazione delle questioni centrali alla nozione di fatto urbano ricca di intuizioni, che aspirano ad un’autonomia, sintetizzate, seppure in modo non sistematico, nella stesura del celebre libro. Si è così cercato di mettere in luce la precisazione nel tempo della nozione di fatto urbano e della sua elaborazione nei molteplici scritti antecedenti la pubblicazione de L’architettura della città, precisando come Rossi, pur costruendo su basi teoriche la nozione di fatto urbano, ne indichi una visione progressiva, ossia un uso operativo sulla città. La ricerca si è proposta come obiettivo di comprendere le radici culturali della nozione di fatto urbano sia tramite un’esplorazione degli interessi di Rossi nel suo percorso formativo sia rispetto alla definizione della struttura materiale del fatto urbano che Rossi individua nelle permanenze e che alimenta nella sua definizione con differenti apporti derivanti da altre discipline. Compito di questa ricerca è stato rileggere criticamente il percorso formativo compiuto da Rossi, a partire dal 1953, sottolinearne gli ambiti innovativi e precisarne i limiti descrittivi che non vedranno mai la determinazione di una nozione esatta, ma piuttosto la strutturazione di una sintesi complessa e ricca di riferimenti ad altri studi. In sintesi la tesi si compone di tre parti: 1. la prima parte, dal titolo “La teoria dei fatti urbani ne L’architettura della città”, analizza il concetto di fatto urbano inserendolo all’interno del più generale contesto teorico contenuto nel libro L’architettura della città. Questo avviene tramite la scomposizione del libro, la concatenazione delle sue argomentazioni e la molteplicità delle fonti esplicitamente citate da Rossi. In questo ambito si precisa la struttura del libro attraverso la rilettura dei riferimenti serviti a Rossi per comporre il suo progetto teorico. Inoltre si ripercorre la sua vita attraverso le varie edizioni, le ristampe, le introduzioni e le illustrazioni. Infine si analizza il ruolo del concetto di fatto urbano nel libro rilevando come sia posto in un rapporto paritetico con il titolo del libro, conseguendone un’accezione di «fatto da osservare» assimilabile all’uso proposto dalla geografia urbana francese dei primi del Novecento. 2. la seconda parte, dal titolo “La formazione della nozione di fatto urbano 1953-66”, è dedicata alla presentazione dell’elaborazione teorica negli scritti di Rossi prima de L’architettura della città, ossia dal 1953 al 1966. Questa parte cerca di descrivere le radici culturali di Rossi, le sue collaborazioni e i suoi interessi ripercorrendo la progressiva definizione della concezione di città nel tempo. Si è analizzato il percorso maturato da Rossi e i documenti scritti fin dagli anni in cui era studente alla Facoltà di Architettura Politecnico di Milano. Emerge un quadro complesso in cui i primi saggi, gli articoli e gli appunti testimoniano una ricerca intellettuale tesa alla costruzione di un sapere sullo sfondo del realismo degli anni Cinquanta. Rossi matura infatti un impegno culturale che lo porta dopo la laurea ad affrontare discorsi più generali sulla città. In particolare la sua importante collaborazione con la rivista Casabella-continuità, con il suo direttore Ernesto Nathan Rogers e tutto il gruppo redazionale segnano il periodo successivo in cui compare l’interesse per la letteratura urbanistica, l’arte, la sociologia, la geografia, l’economia e la filosofia. Seguono poi dal 1963 gli anni di lavoro insieme al gruppo diretto da Carlo Aymonino all’Istituto Universitario di Architettura di Venezia, e in particolare le ricerche sulla tipologia edilizia e la morfologia urbana, che portano Rossi a compiere una sintesi analitica per la fondazione di una teoria della città. Dall’indagine si rileva infatti come gli scritti antecedenti L’architettura della città sviluppano lo studio dei fatti urbani fino ad andare a costituire il nucleo teorico di diversi capitoli del libro. Si racconta così la genesi del libro, la cui scrittura si è svolta nell’arco di due anni, e le aspirazioni che hanno portato quello che era stato concepito come un “manuale d’urbanistica” a divenire quello che Rossi definirà “l’abbozzo di un trattato” per la formulazione di una scienza urbana. 3. la terza parte, dal titolo “La struttura materiale dei fatti urbani: la teoria della permanenza”, indaga monograficamente lo studio della città come un fatto materiale, un manufatto, la cui costruzione è avvenuta nel tempo e del tempo mantiene le tracce. Sul tema della teoria della permanenza è stato importante impostare un confronto con il dibattito vivo negli anni della ricostruzione dopo la guerra intorno ai temi delle preesistenze ambientali nella ricostruzione negli ambienti storici. Sono emersi fin da subito importanti la relazione con Ernesto Nathan Rogers, le discussioni sulle pagine di Casabella-Continuità, la partecipazione ad alcuni dibatti e ricerche. Si è inoltre Rilevato l’uso di diversi termini mutuati dalle tesi filosofiche di alcune personalità come Antonio Banfi e Enzo Paci, poi elaborati dal nucleo redazionale di Casabella-Continuità, di cui faceva parte anche Rossi. Sono così emersi alcuni spostamenti di senso e la formulazione di un vocabolario di termini all’interno della complessa vicenda della cultura architettonica degli anni Cinquanta e Sessanta. 1. Si è poi affrontato questo tema analizzando le forme con cui Rossi presenta la definizione della teoria della permanenza e i contributi desunti da alcuni autori per la costruzione scientifica di una teoria dell’architettura, il cui fine è quello di essere trasmissibile e di offrire strumenti di indagine concreti. Questa ricerca ha permesso di ipotizzare come il lavoro dei geografi francesi della prima metà del XX secolo, e in particolare il contributo più rilevante di Marcel Poëte e di Pierre Lavedan, costituiscono le fonti principali e il campo d’indagine maggiormente esplorato da Rossi per definire la teoria della permanenza e i monumenti. Le permanenze non sono dunque presentate ne L’architettura della città come il “tutto”, ma emergono da un metodo che sceglie di isolare i fatti urbani permanenti, consentendo così di compiere un’ipotesi su “ciò che resta” dopo le trasformazioni continue che operano nella città. Le fonti su cui ho lavorato sono state quelle annunciate da Rossi ne L’architettura della città, e più precisamente i testi nelle edizioni da lui consultate. Anche questo lavoro ha permesso un confronto dei testi che ha fatto emergere ne L’architettura della città l’uso di termini mutuati da linguaggi appartenenti ad altre discipline e quale sia l’uso di concetti estrapolati nella loro interezza. Presupposti metodologici Della formulazione della nozione di fatto urbano si sono indagate l’originalità dell’espressione, le connessioni presunte o contenute negli studi di Rossi sulla città attraverso la raccolta di fonti dirette e indirette che sono andate a formare un notevole corpus di scritti. Le fonti dirette più rilevanti sono state trovare nelle collezioni speciali del Getty Research Institute di Los Angeles in cui sono conservati gli Aldo Rossi Papers, questo archivio comprende materiali inediti dal 1954 al 1988. La natura dei materiali si presenta sotto forma di manoscritti, dattiloscritti, quaderni, documenti ciclostilati, appunti sparsi e una notevole quantità di corrispondenza. Negli Aldo Rossi Papers si trovano anche 32 dei 47 Quaderni Azzurri, le bozze de L’architettura della città e dell’ Autobiografia Scientifica. Per quanto riguarda in particolare L’architettura della città negli Aldo Rossi Papers sono conservati: un quaderno con il titolo “Manuale d’urbanistica, giugno 1963”, chiara prima bozza del libro, degli “Appunti per libro urbanistica estate/inverno 1963”, un quaderno con la copertina rossa datato 20 settembre 1964-8 agosto 1965 e un quaderno con la copertina blu datato 30 agosto 1965-15 dicembre 1965. La possibilità di accedere a questo archivio ha permesso di incrementare la bibliografia relativa agli studi giovanili consentendo di rileggere il percorso culturale in cui Rossi si è formato. E’ così apparsa fondamentale la rivalutazione di alcune questioni relative al realismo socialista che hanno portato a formare un più preciso quadro dei primi scritti di Rossi sullo sfondo di un complesso scenario intellettuale. A questi testi si è affiancata la raccolta delle ricerche universitarie, degli articoli pubblicati su riviste specializzate e degli interventi a dibattiti e seminari. A proposito de L’architettura della città si è raccolta un’ampia letteratura critica riferita sia al testo in specifico che ad una sua collocazione nella storia dell’architettura, mettendo in discussione alcune osservazioni che pongono L’architettura della città come un libro risolutivo e definitivo. Per quanto riguarda il capitolo sulla teoria della permanenza l’analisi è stata svolta a partire dai testi che Rossi stesso indicava ne L’architettura della città rivelando i diversi apporti della letteratura urbanistica francese, e permettendo alla ricerca di precisare le relazioni con alcuni scritti centrali e al contempo colti da Rossi come opportunità per intraprendere l’elaborazione dell’idea di tipo. Per quest’ultima parte si può precisare come Rossi formuli la sua idea di tipo in un contesto culturale dove l’interesse per questo tema era fondamentale. Dunque le fonti che hanno assunto maggior rilievo in quest’ultima fase emergono da un ricco panorama in cui Rossi compie diverse ricerche sia con il gruppo redazionale di Casabella-continuità, sia all’interno della scuola veneziana negli anni Sessanta, ma anche negli studi per l’ILSES e per l’Istituto Nazionale d’Urbanistica. RESEARCH ON THE NOTION OF URBAN ARTIFACT IN THE ARCHITECTURE OF THE CITY BY ALDO ROSSI. Doctoral candidate: Letizia Biondi Tutor: Valter Balducci The present doctoral dissertation deals with the notion of urban artifact that was formulated and presented by Aldo Rossi in his book The Architecture of the City, published in 1966. In The Architecture of the City, the notion of urban artifact is enunciated through a wide range of definitions and forms. In this thesis, a research was done on how the construction of this concept over time was preceded by various studies started in 1953 during the author’s youth, then re-organized and synthesized since 1963 in a manuscript titled “Manual of urban planning” and in two more manuscripts later on. The work of research re-constructed the formulation of the notion of urban artifact through Rossi’s writings. In this sense, the examination of Rossi’s participation in debates, seminars, reviews, university courses or academic researches was of fundamental importance to understand the complexity of a work which is not to be attributed to disciplinary concepts, but to the formulation of a communicable theory. The effort to understand and to explain the notion of urban artifact led to an examination of the meaning used by Rossi to compose The Architecture of the City, which he defines as similar to a treatise. Through this analysis, it emerged that the composition of the book is not directly ascribable to the classical use of editorial writing of a treatise, whose most famous references in the past are the promotion of a correct practice as in the case of Vitruvio’s treatise, or the use of a structure that introduces a new category as in the Alberti case. Contrary to the two founding books, the lack of a global and prescriptive system and the not immediate reference to the writing of a classical treatise are evident in The Architecture of the City. However, the possibility of researching on some unpublished documents allowed to discover that in the writings starting from 1953 the analysis of the questions that are at the core of the notion of urban artifact is rich of intuitions, that aim to autonomy and that would be synthesized, even though not in a systematic way, in his famous book. The attempt was that of highlighting the specification over time of the notion of urban artifact and its elaboration in the various writings preceding the publication of The Architecture of the City. It was also specified that, despite building on theoretical grounds, Rossi indicates a progressive version of the notion of urban artifact, that is a performing use in the city. The present research aims to understand the cultural roots of the notion of urban artifact in two main directions: analyzing, firstly, Rossi’s interests along his formation path and, secondly, the definition of material structure of an urban artifact identified by Rossi in the permanences and enriched by various contributions from other disciplines. The purpose of the present research is to revise the formation path made by Rossi in a critical way, starting by 1953, underlining its innovative aspects and identifying its describing limits, which will never lead to the formulation of an exact notion, but rather to the elaboration of a complex synthesis, enriched by references to other studies. In brief, the thesis is composed of three parts: 1. The first part, titled “The Theory of urban artifacts in The Architecture of the City”, analyzes the concept of urban artifact in the more general theoretical context of the book The Architecture of the City. Such analysis is done by “disassembling” the book, and by linking together the argumentations and the multiplicity of the sources which are explicitly quoted by Rossi. In this context, the book’s structure is defined more precisely through the revision of the references used by Rossi to compose his theoretical project. Moreover, the author’s life is traced back through the various editions, re-printings, introductions and illustrations. Finally, it is specified which role the concept of urban artifact has in the book, pointing out that it is placed in an equal relation with the book’s title; by so doing, the concept of urban artifact gets the new meaning of “fact to be observed”, similar to the use that was suggested by the French urban geography at the beginning of the 20th century. 2. The second part, titled “The formation of the notion of urban artifact 1953-66”, introduces the theoretical elaboration in Rossi’s writings before The Architecture of the City, that is from 1953 to 1966. This part tries to describe Rossi’s cultural roots, his collaborations and his interests, tracing back the progressive definition of his conception of city over time. The analysis focuses on the path followed by Rossi and on the documents that he wrote since the years as a student at the Department of Architecture at the Politecnico in Milan. This leads to a complex scenario of first essays, articles and notes that bear witness to the intellectual research aiming to the construction of a knowledge on the background of the Realism of the 1950s. Rossi develops, in fact, a cultural engagement that leads him after his studies to deal with more general issues about the city. In particular, his important collaboration with the architecture magazine “Casabella-continuità”, with the director Ernesto Nathan Rogers and with the whole redaction staff mark the following period when he starts getting interested in city planning literature, art, sociology, geography, economics and philosophy. Since 1963, Rossi has worked with the group directed by Carlo Aymonino at the “Istituto Universitario di Architettura” (University Institute of Architecture) in Venice, especially researching on building typologies and urban morphology. During these years, Rossi elaborates an analytical synthesis for the formulation of a theory about the city. From the present research, it is evident that the writings preceding The Architecture of the City develop the studies on urban artifacts, which will become theoretical core of different chapters of the book. In conclusion, the genesis of the book is described; written in two years, what was conceived to be an “urban planning manual” became a “treatise draft” for the formulation of an urban science, as Rossi defines it. 3. The third part is titled “The material structure of urban artifacts: the theory of permanence”. This research is made on the study of the city as a material fact, a manufacture, whose construction was made over time, bearing the traces of time. As far as the topic of permanence is concerned, it was also important to draw a comparison with the debate about the issues of environmental pre-existence of re-construction in historical areas, which was very lively during the years of the Reconstruction. Right from the beginning, of fundamental importance were the relationship with Ernesto Nathan Rogers, the discussions on the pages of Casabella-Continuità and the participation to some debates and researches. It is to note that various terms were taken by the philosophical thesis by some personalities such as Antonio Banfi and Enzo Paci, and then re-elaborated by the redaction staff at Casabella-Continuità, which Rossi took part in as well. Through this analysis, it emerged that there were some shifts in meaning and the formulation of a vocabulary of terms within the complex area of the architectonic culture in the 1950s and 1960s. Then, I examined the shapes in which Rossi introduces the definition of the theory of permanence and the references by some authors for the scientific construction of an architecture theory whose aim is being communicable and offering concrete research tools. Such analysis allowed making a hypothesis about the significance for Rossi of the French geographers of the first half of the 20th century: in particular, the work by Marcel Poëte and by Pierre Lavedan is the main source and the research area which Rossi mostly explored to define the theory of permanence and monuments. Therefore, in The Architecture of the City, permanencies are not presented as the “whole”, but they emerge from a method which isolates permanent urban artifacts, in this way allowing making a hypothesis on “what remains” after the continuous transformations made in the city. The sources examined were quoted by Rossi in The Architecture of the City; in particular I analyzed them in the same edition which Rossi referred to. Through such an analysis, it was possible to make a comparison of the texts with one another, which let emerge the use of terms taken by languages belonging to other disciplines in The Architecture of the City and which the use of wholly extrapolated concepts is. Methodological premises As far as the formulation of the notion of urban artifact is concerned, the analysis focuses on the originality of the expression, the connections that are assumed or contained in Rossi’s writings about the city, by collecting direct and indirect sources which formed a significant corpus of writings. The most relevant direct sources were found in the special collections of the Getty Research Institute in Los Angeles, where the “Aldo Rossi Papers” are conserved. This archive contains unpublished material from 1954 to 1988, such as manuscripts, typescripts, notebooks, cyclostyled documents, scraps and notes, and several letters. In the Aldo Rossi Papers there are also 32 out of the 47 Light Blue Notebooks (Quaderni Azzurri), the rough drafts of The Architecture of the City and of the “A Scientific Autobiography”. As regards The Architecture of the City in particular, the Aldo Rossi Papers preserve: a notebook by the title of “Urban planning manual, June, 1963”, which is an explicit first draft of the book; “Notes for urban planning book summer/winter 1963”; a notebook with a red cover dated September 20th, 1964 – August 8th, 1965; and a notebook with a blue cover dated August 30th, 1965 – December 15th, 1965. The possibility of accessing this archive allowed to increase the bibliography related to the youth studies, enabling a revision of the cultural path followed by Rossi’s education. To that end, it was fundamental to re-evaluate some issues linked to the socialist realism which led to a more precise picture of the first writings by Rossi against the background of the intellectual scenario where he formed. In addition to these texts, the collection of university researches, the articles published on specialized reviews and the speeches at debates and seminars were also examined. About The Architecture of the City, a wide-ranging critical literature was collected, related both to the text specifics and to its collocation in the story of architecture, questioning some observations which define The Architecture of the City as a conclusive and definite book. As far as the chapter on the permanence theory is concerned, the analysis started by the texts that Rossi indicated in The Architecture of the City, revealing the different contributions from the French literature on urban planning. This allowed to the present research a more specific definition of the connections to some central writings which, at the same time, were seen by Rossi as an opportunity to start up the elaboration of the idea of type. For this last part, it can be specified that Rossi formulates his idea of type in a cultural context where the interest in this topic was fundamental. Therefore, the sources which played a central role in this final phase emerge from an extensive panorama in which Rossi researched not only with the redaction staff at Casablanca-continuità and within the School of Venice in the 1960s, but also in his studies for the ILSES (Institute of the Region Lombardia for Economics and Social Studies) and for the National Institute of Urban Planning.

Rafael Moneo: la complessità del vuoto. Dalle matrici formali dell'opera di Oteiza e Chillida

The experience of void, essential to the production of forms and to make use them, can be considered as the base of the activities that attend to the formative processes. If void and matter constitutes the basic substances of architecture. Their role in the definition of form, the symbolic value and the constructive methods of it defines the quality of the space. This job inquires the character of space in the architecture of Moneo interpreting the meaning of the void in the Basque culture through the reading of the form matrices in the work of Jorge Oteiza and Eduardo Chillida. In the tie with the Basque culture a reading key is characterized by concurring to put in relation some of the theoretical principles expressed by Moneo on the relationship between place and time, in an unique and specific vision of the space. In the analysis of the process that determines the genesis of the architecture of Moneo emerges a trajectory whose direction is constructed on two pivos: on the one hand architecture like instrument of appropriation of the place, gushed from an acquaintance process who leans itself to the reading of the relations that define the place and of the resonances through which measuring it, on the other hand the architecture whose character is able to represent and to extend the time in which he is conceived, through the autonomy that is conferred to them from values. Following the trace characterized from this hypothesis, that is supported on the theories elaborated from Moneo, surveying deepens the reading of the principles that construct the sculptural work of Oteiza and Chillida, features from a search around the topic of the void and to its expression through the form. It is instrumental to the definition of a specific area that concurs to interpret the character of the space subtended to a vision of the place and the time, affine to the sensibility of Moneo and in some way not stranger to its cultural formation. The years of the academic formation, during which Moneo enters in contact with the Basque artistic culture, seem to be an important period in the birth of that knowledge that will leads him to the formulation of theories tied to the relationship between time, place and architecture. The values expressed through the experimental work of Oteiza and Chillida during years '50 are valid bases to the understanding of such relationships. In tracing a profile of the figures of Oteiza and Chillida, without the pretension that it is exhaustive for the reading of the complex historical period in which they are placed, but with the needs to put the work in a context, I want to be evidenced the important role carried out from the two artists from the Basque cultural area within which Moneo moves its first steps. The tie that approaches Moneo to the Basque culture following the personal trajectory of the formative experience interlaces to that one of important figures of the art and the Spanish architecture. One of the more meaningful relationships is born just during the years of his academic formation, from 1958 to the 1961, when he works like student in the professional office of the architect Francisco Sáenz de Oiza, who was teaching architectural design at the ETSAM. In these years many figures of Basque artists alternated at the professional office of Oiza that enjoys the important support of the manufacturer and maecenas Juan Huarte Beaumont, introduced to he from Oteiza. The tie between Huarte and Oteiza is solid and continuous in the years and it realizes in a contribution to many of the initiatives that makes of Oteiza a forwarder of the Basque culture. In the four years of collaboration with Oiza, Moneo has the opportunity to keep in contact with an atmosphere permeated by a constant search in the field of the plastic art and with figures directly connected to such atmosphere. It’s of a period of great intensity as in the production like in the promotion of the Basque art. The collective “Blanco y Negro”, than is held in 1959 at the Galería Darro to Madrid, is only one of the many times of an exhibition of the work of Oteiza and Chillida. The end of the Fifties is a period of international acknowledgment for Chillida that for Oteiza. The decade of the Fifties consecrates the hypotheses of a mythical past of the Basque people through the spread of the studies carried out in the antecedent years. The archaeological discoveries that join to a context already rich of signs of the prehistoric era, consolidate the knowledge of a strong cultural identity. Oteiza, like Chillida and other contemporary artists, believe in a cosmogonist conception belonging to the Basques, connected to their matriarchal mythological past. The void in its meaning of absence, in the Basque culture, thus as in various archaic and oriental religions, is equivalent to the spiritual fullness as essential condition to the revealing of essence. Retracing the archaic origins of the Basque culture emerges the deep meaning that the void assumes as key element in the religious interpretation of the passage from the life to the death. The symbology becomes rich of meaningful characters who derive from the fact that it is a chthonic cult. A representation of earth like place in which divine manifest itself but also like connection between divine and human, and this manipulation of the matter of which the earth it is composed is the tangible projection of the continuous search of the man towards God. The search of equilibrium between empty and full, that characterizes also the development of the form in architecture, in the Basque culture assumes therefore a peculiar value that returns like constant in great part of the plastic expressions, than in this context seem to be privileged regarding the other expressive forms. Oteiza and Chillida develop two original points of view in the representation of the void through the form. Both use of rigorous systems of rules sensitive to the physics principles and the characters of the matter. The last aim of the Oteiza’s construction is the void like limit of the knowledge, like border between known and unknown. It doesn’t means to reduce the sculptural object to an only allusive dimension because the void as physical and spiritual power is an active void, that possesses that value able to reveal the being through the trace of un-being. The void in its transcendental manifestation acts at the same time from universal and from particular, like in the atomic structure of the matter, in which on one side it constitutes the inner structure of every atom and on the other one it is necessary condition to the interaction between all the atoms. The void can be seen therefore as the action field that concurs the relations between the forms but is also the necessary condition to the same existence of the form. In the construction of Chillida the void represents that counterpart structuring the matter, inborn in it, the element in absence of which wouldn’t be variations neither distinctive characters to define the phenomenal variety of the world. The physics laws become the subject of the sculptural representation, the void are the instrument that concurs to catch up the equilibrium. Chillida dedicate himself to experience the space through the senses, to perceive of the qualities, to tell the physics laws which forge the matter in the form and the form arranges the places. From the artistic experience of the two sculptors they can be transposed, to the architectonic work of Moneo, those matrices on which they have constructed their original lyric expressions, where the void is absolute protagonist. An ambit is defined thus within which the matrices form them drafts from the work of Oteiza and Chillida can be traced in the definition of the process of birth and construction of the architecture of Moneo, but also in the relation that the architecture establishes with the place and in the time. The void becomes instrument to read the space constructed in its relationships that determine the proportions, rhythms, and relations. In this way the void concurs to interpret the architectonic space and to read the value of it, the quality of the spaces constructing it. This because it’s like an instrument of the composition, whose role is to maintain to the separation between the elements putting in evidence the field of relations. The void is that instrument that serves to characterize the elements that are with in the composition, related between each other, but distinguished. The meaning of the void therefore pushes the interpretation of the architectonic composition on the game of the relations between the elements that, independent and distinguished, strengthen themselves in their identity. On the one hand if void, as measurable reality, concurs all the dimensional changes quantifying the relationships between the parts, on the other hand its dialectic connotation concurs to search the equilibrium that regulated such variations. Equilibrium that therefore does not represent an obtained state applying criteria setting up from arbitrary rules but that depends from the intimate nature of the matter and its embodiment in the form. The production of a form, or a formal system that can be finalized to the construction of a building, is indissolubly tied to the technique that is based on the acquaintance of the formal vocation of the matter, and what it also can representing, meaning, expresses itself in characterizing the site. For Moneo, in fact, the space defined from the architecture is above all a site, because the essence of the site is based on the construction. When Moneo speaks about “birth of the idea of plan” like essential moment in the construction process of the architecture, it refers to a process whose complexity cannot be born other than from a deepened acquaintance of the site that leads to the comprehension of its specificity. Specificity arise from the infinite sum of relations, than for Moneo is the story of the oneness of a site, of its history, of the cultural identity and of the dimensional characters that that they are tied to it beyond that to the physical characteristics of the site. This vision is leaned to a solid made physical structure of perceptions, of distances, guideline and references that then make that the process is first of all acquaintance, appropriation. Appropriation that however does not happen for directed consequence because does not exist a relationship of cause and effect between place and architecture, thus as an univocal and exclusive way does not exist to arrive to a representation of an idea. An approach that, through the construction of the place where the architecture acquires its being, searches an expression of its sense of the truth. The proposal of a distinction for areas like space, matter, spirit and time, answering to the issues that scan the topics of the planning search of Moneo, concurs a more immediate reading of the systems subtended to the composition principles, through which is related the recurrent architectonic elements in its planning dictionary. From the dialectic between the opposites that is expressed in the duality of the form, through the definition of a complex element that can mediate between inside and outside as a real system of exchange, Moneo experiences the form development of the building deepening the relations that the volume establishes in the site. From time to time the invention of a system used to answer to the needs of the program and to resolve the dual character of the construction in an only gesture, involves a deep acquaintance of the professional practice. The technical aspect is the essential support to which the construction of the system is indissolubly tied. What therefore arouses interest is the search of the criteria and the way to construct that can reveal essential aspects of the being of the things. The constructive process demands, in fact, the acquaintance of the formative properties of the matter. Property from which the reflections gush on the relations that can be born around the architecture through the resonance produced from the forms. The void, in fact, through the form is in a position to constructing the site establishing a reciprocity relation. A reciprocity that is determined in the game between empty and full and of the forms between each other, regarding around, but also with regard to the subjective experience. The construction of a background used to amplify what is arranged on it and to clearly show the relations between the parts and at the same time able to tie itself with around opening the space of the vision, is a system that in the architecture of Moneo has one of its more effective applications in the use of the platform used like architectonic element. The spiritual force of this architectonic gesture is in the ability to define a place whose projecting intention is perceived and shared with who experience and has lived like some instrument to contact the cosmic forces, in a delicate process that lead to the equilibrium with them, but in completely physical way. The principles subtended to the construction of the form taken from the study of the void and the relations that it concurs, lead to express human values in the construction of the site. The validity of these principles however is tested from the time. The time is what Moneo considers as filter that every architecture is subordinate to and the survival of architecture, or any of its formal characters, reveals them the validity of the principles that have determined it. It manifests thus, in the tie between the spatial and spiritual dimension, between the material and the worldly dimension, the state of necessity that leads, in the construction of the architecture, to establish a contact with the forces of the universe and the intimate world, through a process that translate that necessity in elaboration of a formal system.

Aspetti di vita quotidiana, religiosa, militare e civile in Britannia e lungo il Vallo di Adriano

The dissertation aims to provide an overview of some aspects of everyday life in Roman Britain in general and in the area of Hadrian’s Wall in particular. In a preliminary description, the writer addresses the complex topic related to the genesis of borders as the fulfillment of the expansionist parable of Rome, and as the space manifestation of the sunset of the idea of an imperium sine fine. Then the thesis passes to examine, in subsequent chapters, first the religious theme in its peculiar indigenous component and in the cultural practices triggered by the process of Romanization, secondly the question whether is possible to study everyday life in the northernmost province of the Empire through a discussion of the civilian settlements in proximity to military sites. This issue is drawn especially thanks to the analysis of the so-called Vindolanda tablets, which constitute a valuable evidence of a lively environment both under human and social respect. Before giving an indication of the specific bibliography, the work offers a number of appendices which elaborate part of the information which has been supplied in the previous sections. Mention of the epigraphic repertoires and literary and antiquarian sources is finally made.

L'invention de l'Orthodoxie. Religion et Modernité dans le discours nationaliste roumain du XIX-ème et du XX-ème siècles

Our thesis entitled The Invention of Orthodoxy. Religion and Modernity in Romanian nationalist discourse from the XIXth to the XXth century is intended to be a history of the idea of “Romanianess” which brings together, in a structural as well as in a conceptual dimension, three major themes: Romanian Orthodoxy, Modernity and the Political. Having as premise for the study of the Romanian case the simultaneous genesis of the religious and political communities, from the Middle Ages to Modernity, the purpose of our inquiry is to formulate a theologico-political definition of ‘’Romanian Orthodoxy’’. Thus, within a main theoretical framework that values the contributions of Carl Schmitt, Michel Foucault and Reinhart Koselleck, our analysis of selected texts that go from the 1860’s to the 1940’s tries to answer the question regarding the relationship between Romanian Orthodoxy and Modernity, as well as its reflection upon the political identity and organisation of the Romanian society. Considering the political context of the events that underline our conceptual focus, we consider that the proper answer to our investigation lies within the logic of multiplicity; namely, we refer to a plural Romania which is divided, at the beginning of the XXth century, between Traditionalism and Modernity, between a massive rural, agrarian society and an urban minority elite, striving to single out, in an phenomenological approach, the “Romanian way”. Secondly, we refer to a plural Modernity, which is at the same time social, cultural, religious and political. Thirdly, the logic of multiplicity applies as well in the interpretation of the fractures present within the religious nationalist discourse; namely, the rejection of Orthodoxy during the XIXth century, as it was considered an impediment in Romania’s path to adopting western modernity and later on, starting with the 1930, the restoration of the “Orthodox ethos” as a source of cultural and political values of the Romanian nation.

Studi sull'epigramma scoptico greco

The dissertation is divided into two parts: the first synthesis focused on the definition of the epigram scoptico imperial age, the second analysis concerns the study of the minor poets of Book XI. In the Introduction (I), the attention focuses on the genesis of imperial scoptic epigram: here you try to draw a picture of the satirical Greek literature before the middle of the century AD to identify the debts of the scoptic epigram, especially Lucillius’, in respect of previous authors (from the Middle-up comedy to epigrams of the Crown of Philip), and to emphasize the remoteness of this literary phenomenon from other experiences of ironic and satirical poetry (Catullus). In the chapter on the Themes (II), the study was limited to professional groups and those most targeted (doctors, grammarians, etc..), to that particular type represented by the satire on ethnic groups. The study of minor poets is necessarily preceded by a general discussion on the authors most representative of the greek satiric poetry: Lucillius, Ammianus, Nicarchus and Palladas (III). All the minor poets of the eleventh book, which you can not provide a date, have been regarded by scholars as the ‘poets of Diogenian’: the chapter on Anthologion of Diogenian (IV), which is undergoing critical to the existence (assumed but never proven) of the lost source of Book XI, therefore, serves as an introduction to the commentary of the authors required minors. During the discussion they are not qualified as poets ‘poets of Diogenian’, but are divided into two categories: those included in the string of alphabetically ordered AP XI 388-436 (V), and those who are not part of (VI). Finally, a separate chapter (VII) is devoted to the age-old question of epigrams assigned to Lucian, both in the string of alphabetically ordered epigrams, as well as outside it.

Räumliche Verbreitung geogener Schwermetallgehalte in Böden des Taunus: Einfluss von periglazialer Deckschichtengenese

Während der Glazialphasen kam es in den europäischen Mittelgebirgen bedingt durch extensive solifluidale Massenbewegungen zur Bildung von Deckschichten. Diese Deckschichten repräsentieren eine Mischung verschiedener Substrate, wie anstehendes Ausgangsgestein, äolische Depositionen und lokale Erzgänge. Die räumliche Ausdehnung der Metallkontaminationen verursacht durch kleinräumige Erzgänge wird durch die periglaziale Solifluktion verstärkt. Das Ziel der vorliegenden Untersuchung war a) den Zusammenhang zwischen den Reliefeigenschaften und den Ausprägungen der solifluidalen Deckschichten und Böden aufzuklären, sowie b) mittels Spurenelementgehalte und Blei-Isotopen-Verhältnisse als Eingangsdaten für Mischungsmodelle die Beitrage der einzelnen Substrate zum Ausgangsmaterial der Bodenbildung zu identifizieren und quantifizieren und c) die räumliche Verteilung von Blei (Pb) in Deckschichten, die über Bleierzgänge gewandert sind, untersucht, die Transportweite des erzbürtigen Bleis berechnet und die kontrollierenden Faktoren der Transportweite bestimmt werden. Sechs Transekte im südöstlichen Rheinischen Schiefergebirge, einschließlich der durch periglaziale Solifluktion entwickelten Böden, wurden untersucht. Die bodenkundliche Geländeaufnahme erfolgte nach AG Boden (2005). O, A, B und C-Horizontproben wurden auf ihre Spurenelementgehalte und teilweise auf ihre 206Pb/207Pb-Isotopenverhältnisse analysiert. Die steuernden Faktoren der Verteilung und Eigenschaften periglazialer Deckschichten sind neben der Petrographie, Reliefeigenschaften wie Exposition, Hangneigung, Hangposition und Krümmung. Die Reliefanalyse zeigt geringmächtige Deckschichten in divergenten, konvexen Hangbereichen bei gleichzeitig hohem Skelettgehalt. In konvergent, konkaven Hangbereichen nimmt die Deckschichtenmächtigkeit deutlich zu, bei gleichzeitig zunehmendem Lösslehm- und abnehmendem Skelettgehalt. Abhängig von den Reliefeigenschaften und -positionen reichen die ausgeprägten Bodentypen von sauren Braunerden bis hin zu Pseudogley-Parabraunerden. Des Weiteren kommen holozäne Kolluvien in eher untypischen Reliefpositionen wie langgestreckten, kaum geneigten Hangbereichen oder Mittelhangbereichen vor. Außer für Pb bewegen sich die Spurenelementgehalte im Rahmen niedriger Hintergrundgehalte. Die Pb-Gehalte liegen zwischen 20-135 mg kg-1. Abnehmende Spurenelementgehalte und Isotopensignaturen (206Pb/207Pb-Isotopenverhältnisse) von Pb zeigen, dass nahezu kein Pb aus atmosphärischen Depositionen in die B-Horizonte verlagert wurde. Eine Hauptkomponentenanalyse (PCA) der Spurenelementgehalte hat vier Hauptsubstratquellen der untersuchten B-Horizonte identifiziert (Tonschiefer, Löss, Laacher-See-Tephra [LST] und lokale Pb-Erzgänge). Mittels 3-Komponenten-Mischungsmodell, das Tonschiefer, Löss und LST einschloss, konnten, bis auf 10 Ausreißer, die Spurenelementgehalte aller 120 B-Horizontproben erklärt werden. Der Massenbeitrag des Pb-Erzes zur Substratmischung liegt bei <0,1%. Die räumliche Pb-Verteilung zeigt Bereiche lokaler Pb-Gehaltsmaxima hangaufwärtiger Pb-Erzgänge. Mittels eines 206Pb/207Pb-Isotopenverhältnis-Mischungsmodells konnten 14 Bereiche erhöhter lokaler Pb-Gehaltsmaxima ausgewiesen werden, die 76-100% erzbürtigen Bleis enthalten. Mit Hilfe eines Geographischen Informationssystems wurden die Transportweiten des erzbürtigen Bleis mit 30 bis 110 m bestimmt. Die steuerenden Faktoren der Transportweite sind dabei die Schluffkonzentration und die Vertikalkrümmung. Diese Untersuchung zeigt, dass Reliefeigenschaften und Reliefposition einen entscheidenden Einfluss auf die Ausprägung der Deckschichten und Böden im europäischen Mittelgebirgsbereich haben. Mischungsmodelle in Kombination mit Spurenelementanalysen und Isotopenverhältnissen stellen ein wichtiges Werkzeug zur Bestimmung der Beiträge der einzelnen Glieder in Bodensubstratmischungen dar. Außerdem können lokale Bleierzgänge die natürlichen Pb-Gehalte in Böden, entwickelt in periglazialen Deckschichten der letzten Vereisungsphase (Würm), bis über 100 m Entfernung erhöhen.

Polycyclic aromatic hydrocarbons (PAHs) in river bank soils

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous and found in the atmosphere, aquatic environment, sediments and soils. For environmental risk assessments and the allocation of the polluter it is important to know the PAH sources. PAH contamination sites are usually the result of anthropogenic processes. Three major sources are known: i) petroleum, including crude oil and its refined products and coals (petrogenic PAHs), ii) burning of organic matter (pyrogenic PAHs) and iii) transformation products of natural organic precursors present in the environment (diagenetic processes). In one case elevated PAH concentrations were found in river bank soils when building a retention area along the Mosel River. The source of the PAHs in this area was unclear and required the investigation of possible sources. To evaluate the PAH distribution along the Mosel River, a section of ~ 160 km along the river and a short section along the Saar River were investigated within this study. Concentrations of the Σ16 EPA PAHs were as high as 81 mg kg-1 dry weight (dw). Additionally, coal particles were identified in some soils, which originated from mining activities in the Saarland region. PAH distribution patterns of the 16 EPA PAHs suggest a mainly pyrogenic origin and in some cases a mixture of pyrogenic and petrogenic origin. For a comprehensive investigation five sampling sites were selected. Two sites were located before the confluence of the Mosel and Saar River, one site at the confluence and two sites after the confluence. The examination included typical forensic methods such as PAH distribution patterns of 45 PAHs (including alkylated PAHs), calculation of PAH ratios, determination of PAH alkyl homologues, n-alkanes, principal component analysis (PCA) and coal petrography. The results revealed a mainly pyrogenic source at sampling sites before the confluence of the two rivers. At and after the confluence, a mixture of pyrogenic and petrogenic inputs were present. With the help of coal petrography, coal derived particles could be identified in these soils. Therefore, coal was suggested to be the petrogenic source. It could be shown that sites with diffuse sources of contaminants, like the bank soils of the Mosel River, are difficult to characterize. As previously mentioned for detailed source identifications, the use of various forensic methods is essential. Determination of PAH alkyl homologue series, biomarkers and isotopes are often recommended. Source identification was evaluated using three different methods (i.e. PAH distribution patterns of an extended PAH spectrum, PAH ratios and analyses of n-alkanes). It was assessed if these methods were sufficient for the initial steps in identifying sources of PAHs in selected samples, and if they could be used for decision-making purposes. Point- and non-point sources were identified by applying the three methods and it could be shown that these relatively simple methods are sufficient in determining the primary source. In a last step of this study two soils (one before the confluence of the Mosel and Saar rivers and one after the confluence), and one sediment of the Mosel River were evaluated by investigating the mutagenic potential of the soils and the sediment with a fluctuation version of the Ames-test. The study showed that coal bearing soils at the Mosel River do not exhibit a greater mutagenic potential than other soils or sediments without coal particles.

Studio neuroradiologico (morfologico e funzionale) nei pazienti con Epilessia Frontale Notturna

L'epilessia frontale notturna (EFN) è caratterizzata da crisi motorie che insorgono durante il sonno. Scopo del progetto è studiare le cause fisiopatologiche e morfo-funzionali che sottendono ai fenomeni motori nei pazienti con EFN e identificare alterazioni strutturali e/o metaboliche mediante tecniche avanzate di Risonanza Magnetica (RM). Abbiamo raccolto una casistica di pazienti con EFN afferenti al Centro Epilessia e dei Disturbi del Sonno del Dipartimento di Scienze Neurologiche, Università di Bologna. Ad ogni paziente è stato associato un controllo sano di età (± 5 anni) e sesso corrispondente. Tutti sono stati studiati mediante tecniche avanzate di RM comprendenti Spettroscopia del protone (1H-MRS), Tensore di diffusione ed imaging 3D ad alta risoluzione per analisi morfometriche. In particolare, la 1H-MRS è stata effettuata su due volumi di interesse localizzati nei talami e nel giro del cingolo anteriore. Sono stati inclusi nell’analisi finale 19 pazienti (7 M), età media 34 anni (range 19-50) e 14 controlli (6 M) età media 30 anni (range 19-40). A livello del cingolo anteriore il rapporto della concentrazione di N-Acetil-Aspartato rispetto alla Creatina (NAA/Cr) è risultato significativamente ridotto nei pazienti rispetto ai controlli (p=0,021). Relativamente all’analisi di correlazione, l'analisi tramite modelli di regressione multipla ha evidenziato che il rapporto NAA/Cr nel cingolo anteriore nei pazienti correlava con la frequenza delle crisi (p=0,048), essendo minore nei pazienti con crisi plurisettimanali/plurigiornaliere. Per interpretare il dato ottenuto è possibile solo fare delle ipotesi. L’NAA è un marker di integrità, densità e funzionalità neuronale. E’ possibile che alla base della EFN ci siano alterazioni metaboliche tessutali in precise strutture come il giro del cingolo anteriore. Questo apre nuove possibilità sull’utilizzo di strumenti di indagine basati sull’analisi di biosegnali, per caratterizzare aree coinvolte nella genesi della EFN ancora largamente sconosciute e chiarire ulteriormente l’eziologia di questo tipo di epilessia.

Studio dell'interazione di HIV-1 sulle cellule progenitrici ematopoietiche CD34+ (HPCs)

Oltre alla progressiva perdita dei linfociti T CD4, i pazienti HIV-infetti presentano diverse citopenie periferiche. In particolare l’anemia si riscontra nel 10% dei pazienti asintomatici e nel 92% di quelli con AIDS e la terapia cART non è in grado di risolvere tale problematica. I meccanismi patogenetici alla base di questa citopenia si ritiene che possano riguardare la deregolazione citochinica, il danno alle HPCs, alle cellule in differenziamento e alle cellule stromali. Le cellule progenitrici ematopoietiche CD34+, dopo essere state separate da sangue cordonale e differenziate verso la linea eritroide, sono state trattate con HIV-1 attivo, inattivato al calore e gp120. In prima istanza è stata messa in luce la mancata suscettibilità all’infezione e l’aumento dell’ apoptosi dovuto al legame gp120-CD4/CXCR4 e mediato dal TGF-β1 nelle cellule progenitrici indifferenziate. L’aspetto innovativo di questo studio però si evidenzia esaminando l’effetto di gp120 durante il differenziamento verso la filiera eritrocitaria. Sono stati utilizzati due protocolli sperimentali: nel primo le cellule sono inizialmente trattate per 24 ore con gp120 (o con HIV-1 inattivato al calore) e poi indotte in differenziamento, nel secondo vengono prima differenziate e poi trattate con gp120. Il “priming” negativo determina una apoptosi gp120-indotta molto marcata già dopo 48 ore dal trattamento ed una riduzione del differenziamento. Se tali cellule vengono invece prima differenziate per 24 ore e poi trattate con gp120, nei primi 5 giorni dal trattamento, è presente un aumento di proliferazione e differenziamento, a cui segue un brusco arresto che culmina con una apoptosi molto marcata (anch’essa dipendente dal legame gp120-CD4 e CXCR4 e TGF-β1 dipendente) e con una drastica riduzione del differenziamento. L’insieme dei risultati ha permesso di definire in modo consistente la complessità della genesi dell’anemia in questi pazienti e di poter suggerire nuovi target terapeutici in questi soggetti, già sottoposti a cART.

Funktionelle Analyse der Polaritäts- und Tumorsuppressorgene hugl-1 und hugl-2 mittels siRNA-vermitteltem Gen-Silencing und konditionalem Gen-Knockout

Das menschliche Gen human giant larvae (hugl) ist ein Homolog des hochkonservierten Drosophila Gens lethal giant larvae (lgl), welches in Epithelzellen die Funktion eines neoplastischen Tumorsuppressors und Polaritätsregulators einnimmt. Ein Verlust oder eine verminderte Expression beider Homologe des Gens, hugl-1 und hugl-2, geht einher mit dem Auftreten und der Progression verschiedener epithelialer Tumorerkrankungen wie malignen Melanomen und Brust-, Kolon- oder Lungentumoren. Die exakte Funktion der Homologe Hugl-1 und Hugl-2 bezüglich der Regulation und Aufrechterhaltung der epithelialen Zellpolarität sowie ihre Rolle in der Genese humaner Tumore ist jedoch weitgehend unbekannt. Gänzlich unbekannt ist auch die Bedeutung von Hugl-1 und Hugl-2 als Polaritätsregulatoren für die Ausbildung und den Erhalt der T-Zellmorphologie und -funktion. Ziel der vorliegenden Arbeit war es daher, die Polaritäts- und Tumorsuppressorgene hugl-1 und hugl-2 in funktionellen Analysen mittels siRNA-vermitteltem Gen-Silencing in Epithelzellen und T-Lymphozyten zu charakterisieren. Darüber hinaus wurden die Funktionen und Eigenschaften von mgl-2, dem murinen Homologen von hugl-2, im Cre/loxP-vermittelten konditionalen Knockout Mausmodell in vivo analysiert.rnrnZur Charakterisierung der biologischen Effekte von Hugl-1 und Hugl-2 auf das Wachstumsverhalten, Migration und Invasion von Epithelzellen wurden in dieser Arbeit erfolgreich unterschiedliche shRNA-Expressionskonstrukte generiert sowie Hugl-supprimierte Zelllinien etabliert. In vitro Studien sowie in vivo Tumorigenizitätsanalysen lieferten übereinstimmend Hinweise darauf, dass verminderte Hugl-1- und Hugl-2-Expressionsspiegel eine signifikante Rolle in der Vermittlung invasiver und tumorigener Eigenschaften von Epithelzellen spielen. Dabei rief der Verlust beider Homologe deutlich stärkere Reaktionen hervor als die Suppression eines einzelnen Homologen. Zudem wiesen die Überexpression des Zellzyklusregulators Cyclin D1 sowie die Hyperproliferation von Hugl-1- und/oder Hugl-2-depletierten Epithelzellen auf eine wichtige Rolle der beiden Homologe in der Zellzyklusprogression und Zellproliferation hin. Ein geringer Expressionsstatus von Hugl-1 und -2 schien darüber hinaus mit einer verstärkten Resistenzbildung gegenüber Chemotherapeutika zu korrelieren. Im Rahmen dieser Arbeit konnte weiterhin gezeigt werden, dass die untersuchten T-Lymphozyten nur Hugl-1 exprimieren und dass letzteres notwendig für den F-Aktin-vermittelten Erhalt der T-Zellpolarität und -morphologie ist. Hugl-1-supprimierte, über voneinander unabhängige Signalwege (TCR- oder Chemokinrezeptor) stimulierte T-Lymphozyten wiesen eine bedeutende Störung der Lamellipodien- und Uropodausbildung auf und ließen eine Interaktion von Hugl-1 auf Ebene des F Aktins vermuten. Des Weiteren zeigte sich, dass der Polaritätsregulator Hugl-1 die CD3/TCR-induzierte Zelladhäsion positiv beeinflusst. Die Analyse der T-Zellmigration und -motilität offenbarte in Übereinstimmung dazu die Wichtigkeit von Hugl-1 für die Polarisierung und Migration der T-Zellen sowohl im Chemokingradienten als auch auf mDCs. rnrnFür die Aufklärung der funktionellen Rolle von mgl-2 in vivo wurde in dieser Arbeit eine Tamoxifen-induzierbare, Cre/loxP-vermittelte konditionale Mauslinie generiert und analysiert. Die mgl-2-deletierten Tiere wiesen weder signifikante phänotypische Unterschiede noch Abweichungen in der Organanatomie auf und ließen daher auf eine Kompensation durch das im Darmepithel koexprimierte und möglicherweise funktionell redundante mgl-1 Gen schließen.rn

Untersuchung der physiologischen Rolle des Wilms-Tumorgens 1 (WT1) im Rahmen der Generierung muriner in-vivo-Modelle für die Wilms-Tumorerkrankung

Der Wilms-Tumor ist eine embryonale Tumorerkrankung der Niere, als deren Ursprung Nierenvorläuferzellen des metanephrischen Mesenchyms gelten, deren Differenzierung während der frühen Nephrogenese ausbleibt und aus denen nachfolgend durch eine maligne Transformation Wilms-Tumore entstehen. Zwei Gene, die an der Wilms-Tumorgenese beteiligt zu sein scheinen, sind WT1 (Wilms-Tumorgen 1) und CTNNB1 (Catenin, cadherin-associated protein, beta 1). Während WT1 u.a. die Differenzierung des metanephrischen Mesenchyms steuert, begünstigen aktivierende Mutationen von CTNNB1 und eine dadurch bedingte Akkumulation seines Proteins β-Catenin die Tumorgenese vieler Organe. So verwundert es nicht, dass eine alleinige heterozygote Keimbahnmutation von WT1, die einen dominant-negativen Effekt auf funktionsfähiges WT1 ausübt, häufig zur Entstehung von Wilms-Tumoren in Patienten mit Denys-Drash-Syndrom (DDS) führt, sowie in etwa 15 % aller sporadischen Wilms-Tumore WT1 und CTNNB1 mutiert sind.rnDer Mechanismus der Entstehung von Wilms-Tumoren ist weitgehend unbekannt, was u.a. daran liegt, dass homozygote Wt1-Mutationen in der Maus embryonal (~ Tag 13,5 d.p.c.) letal sind. In der vorliegenden Arbeit sollten daher mit Hilfe einer Wt1 k.o.-Effektormaus (WE2) vier murine konditional reversible Wilms-Tumor-Modelle auf Basis des Tet off-Systems hergestellt werden. Dadurch lag in den zu generierenden Tieren Wt1 durch die Integration des WE2-Transgens zwar nur heterozygot mutiert vor, doch durch den endogenen Wt1-Promotor des Transgens sollte es zur zeitlichen und räumlichen Wt1-analogen Expression eines tetrazyklinabhängigen Transaktivators (tTA) kommen, der ohne die Gabe von Doxycyclin Tet-regulierbare Transgene in Wt1-exprimierenden Zellen aktivieren kann, die einen positiven Einfluss auf die Wilms-Tumorgenese haben könnten. So sollte durch das WE2 DDS-Modell ein DDS simuliert werden und es in Tieren der Modelle WE2 TC bCat∆Ex3, WE2 LC bCat∆Ex3 und WE2 Wnt1 zur Akkumulation von β-Catenin in Wt1-exprimierenden Nierenvorläuferzellen kommen, so dass deren Differenzierung ausbleibt und es durch eine maligne Transformation zur Entstehung eines Wilms-Tumors kommt.rnrnMit Hilfe von histologischen Analysen an entsprechenden Responder-Linien konnte zunächst gezeigt werden, dass die embryonale und adulte Expressionsdomäne des WE2-Effektors mit der von endogenen Wt1 übereinstimmt. Gleichzeitig wurden aber auch neue Expressionsorte von Wt1 nachgewiesen. So konnte die Expression des WE2-Effektors z.B. im Endothel der dorsalen Aorta detektiert werden, der als Entstehungsort von hämatopoetischen Stammzellen gilt. Anschließende hier vorgestellte Experimente zeigten, dass Wt1 direkt an diesem Prozess beteiligt ist und belegten eine noch nicht beschriebene Funktion von Wt1 in der frühen Hämatopoese.rnEs war jedoch mit keinem System möglich, eine Wilms-Tumorerkrankung zu simulieren. Während Tiere des WE2 DDS-Modells trotz nachweisbarer Induktion keinen Phänotyp aufwiesen, war wohl in den anderen Modellen eine konstitutive β-Catenin-Aktivierung in der Frühschwangerschaft nicht mit dem embryonalen Überleben vereinbar. Dabei schienen alle tripeltransgenen bzw. doppeltransgenen Embryonen, in denen durch einen frühen Doxycyclinentzug die Entstehung von Wilms-Tumoren möglich gewesen wäre, intrauterin zu sterben. Wurde dagegen Doxycyclin erst in der dritten Lebenswoche entzogen, so entwickelten die Tiere durch eine Wt1-vermittelte β-Catenin-Aktivierung Granulosazelltumore, polyzystische Nieren und Veränderungen der Hoden. Da alle diese organischen Veränderungen während der prä- bis frühen postnatalen Phase induziert wurden, schien die Doxycyclinmenge nicht auszureichen, um eine β-Catenin-Aktivierung zu verhindern. Es hätte also auch zur Entstehung von Wilms-Tumoren kommen können, so dass diese Ergebnisse darauf hinweisen, dass eine β-Catenin-Aktivierung wahrscheinlich nicht der physiologisch entscheidende Schritt bei der Entstehung eines Wilms-Tumors ist.rnrnDie Charakterisierung der WE2-Effektormaus und die Herstellung und Analysen der Systeme geben damit Einblick in die WT1- bzw. WT1/CTNNB1-assoziierte Wilms-Tumorgenese und ermöglichen die weitere Erforschung von Granulosazelltumoren, polyzystsischen Nieren, Veränderungen von Hoden und der Rolle von WT1 in der frühen Hämatopoese.rn