Genes involved in thoracic exoskeleton formation during the pupal-to-adult molt in a social insect model, Apis mellifera

Background: The insect exoskeleton provides shape, waterproofing, and locomotion via attached somatic muscles. The exoskeleton is renewed during molting, a process regulated by ecdysteroid hormones. The holometabolous pupa transforms into an adult during the imaginal molt, when the epidermis synthe3sizes the definitive exoskeleton that then differentiates progressively. An important issue in insect development concerns how the exoskeletal regions are constructed to provide their morphological, physiological and mechanical functions. We used whole-genome oligonucleotide microarrays to screen for genes involved in exoskeletal formation in the honeybee thoracic dorsum. Our analysis included three sampling times during the pupal-to-adult molt, i.e., before, during and after the ecdysteroid-induced apolysis that triggers synthesis of the adult exoskeleton. Results: Gene ontology annotation based on orthologous relationships with Drosophila melanogaster genes placed the honeybee differentially expressed genes (DEGs) into distinct categories of Biological Process and Molecular Function, depending on developmental time, revealing the functional elements required for adult exoskeleton formation. Of the 1,253 unique DEGs, 547 were upregulated in the thoracic dorsum after apolysis, suggesting induction by the ecdysteroid pulse. The upregulated gene set included 20 of the 47 cuticular protein (CP) genes that were previously identified in the honeybee genome, and three novel putative CP genes that do not belong to a known CP family. In situ hybridization showed that two of the novel genes were abundantly expressed in the epidermis during adult exoskeleton formation, strongly implicating them as genuine CP genes. Conserved sequence motifs identified the CP genes as members of the CPR, Tweedle, Apidermin, CPF, CPLCP1 and Analogous-to-Peritrophins families. Furthermore, 28 of the 36 muscle-related DEGs were upregulated during the de novo formation of striated fibers attached to the exoskeleton. A search for cis-regulatory motifs in the 5′-untranslated region of the DEGs revealed potential binding sites for known transcription factors. Construction of a regulatory network showed that various upregulated CP- and muscle-related genes (15 and 21 genes, respectively) share common elements, suggesting co-regulation during thoracic exoskeleton formation. Conclusions: These findings help reveal molecular aspects of rigid thoracic exoskeleton formation during the ecdysteroid-coordinated pupal-to-adult molt in the honeybee.

Exercising the self : On the Role of Exercise, Gender and Culture in Physical Self-Perceptions

In modern society, individuals constantly pass judgments on their own body and physical competence as well as that of other people. All too often, the verdict is less favourable. For the person, these physical self-perceptions (PSP) may negatively affect global self-esteem, identity, and general mental well being. The overall aim of this thesis is to examine primarily the role that exercise, but also the roles that gender and culture, play in the formation of PSP. In Study I, using confirmatory factor analyses, strong support for the validity of a first-order, and a second-order hierarchical and multidimensional model of the Physical Self-Perception Profile (PSPP: Fox & Corbin, 1989) was found across three national samples (Great Britain, Sweden and Turkey) of university students. Cross-cultural differences were detected, with the British sample demonstrating higher latent means on all PSPP subdomains except for the physical condition subdomain (Condition), than the Swedish and Turkish samples. In Study II, a higher self-reported exercise frequency was associated with more positive PSP (in particular for Condition) and more importance attributed to PSP in Swedish university students. Males demonstrated higher overall PSPP-scores than females. In Study III, a true-experimental design with randomisation into an intervention and a control group was adopted. Strong support for the effects of an empowerment-based exercise intervention programme on PSP and social physique anxiety (SPA) over six months for adolescent girls was found. The relations of exercise, gender and culture with PSP, SPA and self-esteem are discussed from the standpoints of a variety of theoretical models (the EXSEM-model), and frameworks (self-presentation and objectification theory). The two theories of self-enhancement and skill-development are examined with regard to the direction of the exercise-physical self relationship and motivation for exercise. Arguments for the relevance of exercise and PSP for practitioners in promoting general mental well-being and preventing modern-day diseases are outlined.

L'eredità di Aldo Rossi nella Penisola Iberica. La costruzione della città e il progetto contemporaneo

Descrizione, tema e obiettivi della ricerca La ricerca si propone lo studio delle possibili influenze che la teoria di Aldo Rossi ha avuto sulla pratica progettuale nella Penisola Iberica, intende quindi affrontare i caratteri fondamentali della teoria che sta alla base di un metodo progettuale ed in particolar modo porre l'attenzione alle nuove costruzioni quando queste si confrontano con le città storiche. Ha come oggetto principale lo studio dei documenti, saggi e scritti riguardanti il tema della costruzione all'interno delle città storiche. Dallo studio di testi selezionati di Aldo Rossi sulla città si vuole concentrare l'attenzione sull'influenza che tale teoria ha avuto nei progetti della Penisola Iberica, studiare come è stata recepita e trasmessa successivamente, attraverso gli scritti di autori spagnoli e come ha visto un suo concretizzarsi poi nei progetti di nuove costruzioni all'interno delle città storiche. Si intende restringere il campo su un periodo ed un luogo precisi, Spagna e Portogallo a partire dagli anni Settanta, tramite la lettura di un importante evento che ha ufficializzato il contatto dell'architetto italiano con la Penisola Iberica, quale il Seminario di Santiago de Compostela tenutosi nel 1976. Al Seminario parteciparono numerosi architetti che si confrontarono su di un progetto per la città di Santiago e furono invitati personaggi di fama internazionale a tenere lezioni introduttive sul tema di dibattito in merito al progetto e alla città storica. Il Seminario di Santiago si colloca in un periodo storico cruciale per la Penisola Iberica, nel 1974 cade il regime salazarista in Portogallo e nel 1975 cade il regime franchista in Spagna ed è quindi di rilevante importanza capire il legame tra l'architettura e la nuova situazione politica. Dallo studio degli interventi, dei progetti che furono prodotti durante il Seminario, della relazione tra questo evento ed il periodo storico in cui esso va contestualizzato, si intende giungere alla individuazione delle tracce della reale presenza di tale eredità. Presupposti metodologici. Percorso e strumenti di ricerca La ricerca può quindi essere articolata in distinte fasi corrispondenti per lo più ai capitoli in cui si articola la tesi: una prima fase con carattere prevalentemente storica, di ricerca del materiale per poter definire il contesto in cui si sviluppano poi le vicende oggetto della tesi; una seconda fase di impronta teorica, ossia di ricerca bibliografica del materiale e delle testimonianze che provvedono alla definizione della reale presenza di effetti scaturiti dai contatti tra Rossi e la Penisola Iberica, per andare a costruire una eredità ; una terza fase che entra nel merito della composizione attraverso lo studio e la verifica delle prime due parti, tramite l'analisi grafica applicata ad uno specifico esempio architettonico selezionato; una quarta fase dove il punto di vista viene ribaltato e si indaga l'influenza dei luoghi visitati e dei contatti intrattenuti con alcuni personaggi della Penisola Iberica sull'architettura di Rossi, ricercandone i riferimenti. La ricerca è stata condotta attraverso lo studio di alcuni eventi selezionati nel corso degli anni che si sono mostrati significativi per l'indagine, per la risonanza che hanno avuto sulla storia dell'architettura della Penisola. A questo scopo si sono utilizzati principalmente tre strumenti: lo studio dei documenti, le pubblicazioni e le riviste prodotte in Spagna, gli scritti di Aldo Rossi in merito, e la testimonianza diretta attraverso interviste di personaggi chiave. La ricerca ha prodotto un testo suddiviso per capitoli che rispetta l'organizzazione in fasi di lavoro. A seguito di determinate condizioni storiche e politiche, studiate nella ricerca a supporto della tesi espressa, nella Penisola Iberica si è verificato il diffondersi della necessità e del desiderio di guardare e prendere a riferimento l'architettura europea e in particolar modo quella italiana. Il periodo sul quale viene focalizzata l'attenzione ha inizio negli anni Sessanta, gli ultimi prima della caduta delle dittature, scenario dei primi viaggi di Aldo Rossi nella Penisola Iberica. Questi primi contatti pongono le basi per intense e significative relazioni future. Attraverso l'approfondimento e la studio dei materiali relativi all'oggetto della tesi, si è cercato di mettere in luce il contesto culturale, l'attenzione e l'interesse per l'apertura di un dibattito intorno all'architettura, non solo a livello nazionale, ma europeo. Ciò ha evidenziato il desiderio di innescare un meccanismo di discussione e scambio di idee, facendo leva sull'importanza dello sviluppo e ricerca di una base teorica comune che rende coerente i lavori prodotti nel panorama architettonico iberico, seppur ottenendo risultati che si differenziano gli uni dagli altri. E' emerso un forte interesse per il discorso teorico sull'architettura, trasmissibile e comunicabile, che diventa punto di partenza per un metodo progettuale. Ciò ha reso palese una condivisione di intenti e l'assunzione della teoria di Aldo Rossi, acquisita, diffusa e discussa, attraverso la pubblicazione dei suoi saggi, la conoscenza diretta con l'architetto e la sua architettura, conferenze, seminari, come base teorica su cui fondare il proprio sapere architettonico ed il processo metodologico progettuale da applicare di volta in volta negli interventi concreti. Si è giunti così alla definizione di determinati eventi che hanno permesso di entrare nel profondo della questione e di sondare la relazione tra Rossi e la Penisola Iberica, il materiale fornito dallo studio di tali episodi, quali il I SIAC, la diffusione della rivista "2C. Construccion de la Ciudad", la Coleccion Arquitectura y Critica di Gustavo Gili, hanno poi dato impulso per il reperimento di una rete di ulteriori riferimenti. E' stato possibile quindi individuare un gruppo di architetti spagnoli, che si identificano come allievi del maestro Rossi, impegnato per altro in quegli anni nella formazione di una Scuola e di un insegnamento, che non viene recepito tanto nelle forme, piuttosto nei contenuti. I punti su cui si fondano le connessioni tra l'analisi urbana e il progetto architettonico si centrano attorno due temi di base che riprendono la teoria esposta da Rossi nel saggio L'architettura della città : - relazione tra l'area-studio e la città nella sua globalità, - relazione tra la tipologia edificatoria e gli aspetti morfologici. La ricerca presentata ha visto nelle sue successive fasi di approfondimento, come si è detto, lo sviluppo parallelo di più tematiche. Nell'affrontare ciascuna fase è stato necessario, di volta in volta, operare una verifica delle tappe percorse precedentemente, per mantenere costante il filo del discorso col lavoro svolto e ritrovare, durante lo svolgimento stesso della ricerca, gli elementi di connessione tra i diversi episodi analizzati. Tale operazione ha messo in luce talvolta nodi della ricerca rimasti in sospeso che richiedevano un ulteriore approfondimento o talvolta solo una rivisitazione per renderne possibile un più proficuo collegamento con la rete di informazioni accumulate. La ricerca ha percorso strade diverse che corrono parallele, per quanto riguarda il periodo preso in analisi: - i testi sulla storia dell'architettura spagnola e la situazione contestuale agli anni Settanta - il materiale riguardante il I SIAC - le interviste ai partecipanti al I SIAC - le traduzioni di Gustavo Gili nella Coleccion Arquitectura y Critica - la rivista "2C. Construccion de la Ciudad" Esse hanno portato alla luce una notevole quantità di tematiche, attraverso le quali, queste strade vengono ad intrecciarsi e a coincidere, verificando l'una la veridicità dell'altra e rafforzandone il valore delle affermazioni. Esposizione sintetica dei principali contenuti esposti dalla ricerca Andiamo ora a vedere brevemente i contenuti dei singoli capitoli. Nel primo capitolo Anni Settanta. Periodo di transizione per la Penisola Iberica si è cercato di dare un contesto storico agli eventi studiati successivamente, andando ad evidenziare gli elementi chiave che permettono di rintracciare la presenza della predisposizione ad un cambiamento culturale. La fase di passaggio da una condizione di chiusura rispetto alle contaminazioni provenienti dall'esterno, che caratterizza Spagna e Portogallo negli anni Sessanta, lascia il posto ad un graduale abbandono della situazione di isolamento venutasi a creare intorno al Paese a causa del regime dittatoriale, fino a giungere all'apertura e all'interesse nei confronti degli apporti culturali esterni. E' in questo contesto che si gettano le basi per la realizzazione del I Seminario Internazionale di Architettura Contemporanea a Santiago de Compostela, del 1976, diretto da Aldo Rossi e organizzato da César Portela e Salvador Tarragó, di cui tratta il capitolo secondo. Questo è uno degli eventi rintracciati nella storia delle relazioni tra Rossi e la Penisola Iberica, attraverso il quale è stato possibile constatare la presenza di uno scambio culturale e l'importazione in Spagna delle teorie di Aldo Rossi. Organizzato all'indomani della caduta del franchismo, ne conserva una reminescenza formale. Il capitolo è organizzato in tre parti, la prima si occupa della ricostruzione dei momenti salienti del Seminario Proyecto y ciudad historica, dagli interventi di architetti di fama internazionale, quali lo stesso Aldo Rossi, Carlo Aymonino, James Stirling, Oswald Mathias Ungers e molti altri, che si confrontano sul tema delle città storiche, alle giornate seminariali dedicate all’elaborazione di un progetto per cinque aree individuate all’interno di Santiago de Compostela e quindi dell’applicazione alla pratica progettuale dell’inscindibile base teorica esposta. Segue la seconda parte dello stesso capitolo riguardante La selezione di interviste ai partecipanti al Seminario. Esso contiene la raccolta dei colloqui avuti con alcuni dei personaggi che presero parte al Seminario e attraverso le loro parole si è cercato di approfondire la materia, in particolar modo andando ad evidenziare l’ambiente culturale in cui nacque l’idea del Seminario, il ruolo avuto nella diffusione della teoria di Aldo Rossi in Spagna e la ripercussione che ebbe nella pratica costruttiva. Le diverse interviste, seppur rivolte a persone che oggi vivono in contesti distanti e che in seguito a questa esperienza collettiva hanno intrapreso strade diverse, hanno fatto emergere aspetti comuni, tale unanimità ha dato ancor più importanza al valore di testimonianza offerta. L’elemento che risulta più evidente è il lascito teorico, di molto prevalente rispetto a quello progettuale che si è andato mescolando di volta in volta con la tradizione e l’esperienza dei cosiddetti allievi di Aldo Rossi. Negli stessi anni comincia a farsi strada l’importanza del confronto e del dibattito circa i temi architettonici e nel capitolo La fortuna critica della teoria di Aldo Rossi nella Penisola Iberica è stato affrontato proprio questo rinnovato interesse per la teoria che in quegli anni si stava diffondendo. Si è portato avanti lo studio delle pubblicazioni di Gustavo Gili nella Coleccion Arquitectura y Critica che, a partire dalla fine degli anni Sessanta, pubblica e traduce in lingua spagnola i più importanti saggi di architettura, tra i quali La arquitectura de la ciudad di Aldo Rossi, nel 1971, e Comlejidad y contradiccion en arquitectura di Robert Venturi nel 1972. Entrambi fondamentali per il modo di affrontare determinate tematiche di cui sempre più in quegli anni si stava interessando la cultura architettonica iberica, diventando così ¬ testi di riferimento anche nelle scuole. Le tracce dell’influenza di Rossi sulla Penisola Iberica si sono poi ricercate nella rivista “2C. Construccion de la Ciudad” individuata come strumento di espressione di una teoria condivisa. Con la nascita nel 1972 a Barcellona di questa rivista viene portato avanti l’impegno di promuovere la Tendenza, facendo riferimento all’opera e alle idee di Rossi ed altri architetti europei, mirando inoltre al recupero di un ruolo privilegiato dell’architettura catalana. A questo proposito sono emersi due fondamentali aspetti che hanno legittimato l’indagine e lo studio di questa fonte: - la diffusione della cultura architettonica, il controllo ideologico e di informazione operato dal lavoro compiuto dalla rivista; - la documentazione circa i criteri di scelta della redazione a proposito del materiale pubblicato. E’ infatti attraverso le pubblicazioni di “2C. Construccion de la Ciudad” che è stato possibile il ritrovamento delle notizie sulla mostra Arquitectura y razionalismo. Aldo Rossi + 21 arquitectos españoles, che accomuna in un’unica esposizione le opere del maestro e di ventuno giovani allievi che hanno recepito e condiviso la teoria espressa ne “L’architettura della città”. Tale mostra viene poi riproposta nella Sezione Internazionale di Architettura della XV Triennale di Milano, la quale dedica un Padiglione col titolo Barcelona, tres epocas tres propuestas. Dalla disamina dei progetti presentati è emerso un interessante caso di confronto tra le Viviendas para gitanos di César Portela e la Casa Bay di Borgo Ticino di Aldo Rossi, di cui si è occupato l’ultimo paragrafo di questo capitolo. Nel corso degli studi è poi emerso un interessante risvolto della ricerca che, capovolgendone l’oggetto stesso, ne ha approfondito gli aspetti cercando di scavare più in profondità nell’analisi della reciproca influenza tra la cultura iberica e Aldo Rossi, questa parte, sviscerata nell’ultimo capitolo, La Penisola Iberica nel “magazzino della memoria” di Aldo Rossi, ha preso il posto di quello che inizialmente doveva presentarsi come il risvolto progettuale della tesi. Era previsto infatti, al termine dello studio dell’influenza di Aldo Rossi sulla Penisola Iberica, un capitolo che concentrava l’attenzione sulla produzione progettuale. A seguito dell’emergere di un’influenza di carattere prettamente teorica, che ha sicuramente modificato la pratica dal punto di vista delle scelte architettoniche, senza però rendersi esplicita dal punto di vista formale, si è preferito, anche per la difficoltà di individuare un solo esempio rappresentativo di quanto espresso, sostituire quest’ultima parte con lo studio dell’altra faccia della medaglia, ossia l’importanza che a sua volta ha avuto la cultura iberica nella formazione della collezione dei riferimenti di Aldo Rossi. L’articolarsi della tesi in fasi distinte, strettamente connesse tra loro da un filo conduttore, ha reso necessari successivi aggiustamenti nel percorso intrapreso, dettati dall’emergere durante la ricerca di nuovi elementi di indagine. Si è pertanto resa esplicita la ricercata eredità di Aldo Rossi, configurandosi però prevalentemente come un’influenza teorica che ha preso le sfumature del contesto e dell’esperienza personale di chi se ne è fatto ricevente, diventandone così un continuatore attraverso il proprio percorso autonomo o collettivo intrapreso in seguito. Come suggerisce José Charters Monteiro, l’eredità di Rossi può essere letta attraverso tre aspetti su cui si basa la sua lezione: la biografia, la teoria dell’architettura, l’opera. In particolar modo per quanto riguarda la Penisola Iberica si può parlare dell’individuazione di un insegnamento riferito alla seconda categoria, i suoi libri di testo, le sue partecipazioni, le traduzioni. Questo è un lascito che rende possibile la continuazione di un dibattito in merito ai temi della teoria dell’architettura, della sue finalità e delle concrete applicazioni nelle opere, che ha permesso il verificarsi di una apertura mentale che mette in relazione l’architettura con altre discipline umanistiche e scientifiche, dalla politica, alla sociologia, comprendendo l’arte, le città la morfologia, la topografia, mediate e messe in relazione proprio attraverso l’architettura.

A candidate gene approach to identify DNA markers associated with meat quality and production traits: investigation of several cathepsin genes in Italian heavy pigs.

The cathepsin enzymes represent an important family of lysosomal proteinases with a broad spectrum of functions in many, if not in all, tissues and cell types. In addition to their primary role during the normal protein turnover, they possess highly specific proteolytic activities, including antigen processing in the immune response and a direct role in the development of obesity and tumours. In pigs, the involvement of cathepsin enzymes in proteolytic processes have important effects during the conversion of muscle to meat, due to their influence on meat texture and sensory characteristics, mainly in seasoned products. Their contribution is fundamental in flavour development of dry-curing hams. However, several authors have demonstrated that high cathepsin activity, in particular of cathepsin B, is correlated to defects of these products, such as an excessive meat softness together with abnormal free tyrosine content, astringent or metallic aftertastes and formation of a white film on the cut surface. Thus, investigation of their genetic variability could be useful to identify DNA markers associated with these dry cured hams parameters, but also with meat quality, production and carcass traits in Italian heavy pigs. Unfortunately, no association has been found between cathepsin markers and meat quality traits so far, in particular with cathepsin B activity, suggesting that other genes, besides these, affect meat quality parameters. Nevertheless, significant associations were observed with several carcass and production traits in pigs. A recent study has demonstrated that different single nucleotide polymorphisms (SNPs) localized in cathepsin D (CTSD), F (CTSF), H and Z genes were highly associated with growth, fat deposition and production traits in an Italian Large White pig population. The aim of this thesis was to confirm some of these results in other pig populations and identify new cathepsin markers in order to evaluate their effects on cathepsin activity and other production traits. Furthermore, starting from the data obtained in previous studies on CTSD gene, we also analyzed the known polymorphism located in the insulin-like growth factor 2 gene (IGF2 intron3-g.3072G>A). This marker is considered the causative mutation for the quantitative trait loci (QTL) affecting muscle mass and fat deposition in pigs. Since IGF2 maps very close to CTSD on porcine chromosome (SSC) 2, we wanted to clarify if the effects of the CTSD marker were due to linkage disequilibrium with the IGF2 intron3-g.3072G>A mutation or not. In the first chapter, we reported the results from these two SSC2 gene markers. First of all, we evaluated the effects of the IGF2 intron3-g.3072G>A polymorphism in the Italian Large White breed, for which no previous studies have analysed this marker. Highly significant associations were identified with all estimated breeding values for production and carcass traits (P<0.00001), while no effects were observed for meat quality traits. Instead, the IGF2 intron3-g.3072G>A mutation did not show any associations with the analyzed traits in the Italian Duroc pigs, probably due to the low level of variability at this polymorphic site for this breed. In the same Duroc pig population, significant associations were obtained for the CTSD marker for all production and carcass traits (P < 0.001), after excluding possible confounding effects of the IGF2 mutation. The effects of the CTSD g.70G>A polymorphism were also confirmed in a group of Italian Large White pigs homozygous for the IGF2 intron3-g.3072G allele G (IGF2 intron3-g.3072GG) and by haplotype analysis between the markers of the two considered genes. Taken together, all these data indicated that the IGF2 intron3-g.3072G>A mutation is not the only polymorphism affecting fatness and muscle deposition in pigs. In the second chapter, we reported the analysis of two new SNPs identified in cathepsin L (CTSL) and cathepsin S (CTSS) genes and the association results with meat quality parameters (including cathepsin B activity) and several production traits in an Italian Large White pig population. Allele frequencies of these two markers were evaluated in 7 different pig breeds. Furthermore, we mapped using a radiation hybrid panel the CTSS gene on SSC4. Association studies with several production traits, carried out in 268 Italian Large White pigs, indicated positive effects of the CTSL polymorphism on average daily gain, weight of lean cuts and backfat thickness (P<0.05). The results for these latter traits were also confirmed using a selective genotype approach in other Italian Large White pigs (P<0.01). In the 268 pig group, the CTSS polymorphism was associated with feed:gain ratio and average daily gain (P<0.05). Instead, no association was observed between the analysed markers and meat quality parameters. Finally, we wanted to verify if the positive results obtained for the cathepsin L and S markers and for other previous identified SNPs (cathepsin F, cathepsin Z and their inhibitor cystatin B) were confirmed in the Italian Duroc pig breed (third chapter). We analysed them in two groups of Duroc pigs: the first group was made of 218 performance-tested pigs not selected by any phenotypic criteria, the second group was made of 100 Italian Duroc pigs extreme and divergent for visible intermuscular fat trait. In the first group, the CTSL polymorphism was associated with weight of lean cuts (P<0.05), while suggestive associations were obtained for average daily gain and backfat thickness (P<0.10). Allele frequencies of the CTSL gene marker also differed positively among the visible intermuscular extreme tails. Instead, no positive effects were observed for the other DNA markers on the analysed traits. In conclusion, in agreement with the present data and for the biological role of these enzymes, the porcine CTSD and CTSL markers: a) may have a direct effect in the biological mechanisms involved in determining fat and lean meat content in pigs, or b) these markers could be very close to the putative functional mutation(s) present in other genes. These findings have important practical applications, in particular the CTSD and CTSL mutations could be applied in a marker assisted selection (MAS) both in the Italian Large White and Italian Duroc breeds. Marker assisted selection could also increase in efficiency by adding information from the cathepsin S genotype, but only in the Italian Large White breed.

Investigations on formation and specification of neural precursor cells in the central nervous system of the Drosophila melanogaster embryo

Investigations on formation and specification of neural precursor cells in the central nervous system of the Drosophila melanogaster embryoSpecification of a unique cell fate during development of a multicellular organism often is a function of its position. The Drosophila central nervous system (CNS) provides an ideal system to dissect signalling events during development that lead to cell specific patterns. Different cell types in the CNS are formed from a relatively few precursor cells, the neuroblasts (NBs), which delaminate from the neurogenic region of the ectoderm. The delamination occurs in five waves, S1-S5, finally leading to a subepidermal layer consisting of about 30 NBs, each with a unique identity, arranged in a stereotyped spatial pattern in each hemisegment. This information depends on several factors such as the concentrations of various morphogens, cell-cell interactions and long range signals present at the position and time of its birth. The early NBs, delaminating during S1 and S2, form an orthogonal array of four rows (2/3,4,5,6/7) and three columns (medial, intermediate, and lateral) . However, the three column and four row-arrangement pattern is only transitory during early stages of neurogenesis which is obscured by late emerging (S3-S5) neuroblasts (Doe and Goodman, 1985; Goodman and Doe, 1993). Therefore the aim of my study has been to identify novel genes which play a role in the formation or specification of late delaminating NBs.In this study the gene anterior open or yan was picked up in a genetic screen to identity novel and yet unidentified genes in the process of late neuroblast formation and specification. I have shown that the gene yan is responsible for maintaining the cells of the neuroectoderm in an undifferentiated state by interfering with the Notch signalling mechanism. Secondly, I have studied the function and interactions of segment polarity genes within a certain neuroectodermal region, namely the engrailed (en) expressing domain, with regard to the fate specification of a set of late neuroblasts, namely NB 6-4 and NB 7-3. I have dissected the regulatory interaction of the segment polarity genes wingless (wg), hedgehog (hh) and engrailed (en) as they maintain each other’s expression to show that En is a prerequisite for neurogenesis and show that the interplay of the segmentation genes naked (nkd) and gooseberry (gsb), both of which are targets of wingless (wg) activity, leads to differential commitment of NB 7-3 and NB 6-4 cell fate. I have shown that in the absence of either nkd or gsb one NB fate is replaced by the other. However, the temporal sequence of delamination is maintained, suggesting that formation and specification of these two NBs are under independent control.

Formal planning rationality in public sector professional work: between discourse and practice

Although rational models of formal planning have been seriously criticized by strategy literature, they not only remain a widely used organizational practice in private firms, but they have increasingly been entering public, professional organizations too, as part of public sector managerial reforms. This research addresses this apparent paradox, exploring the meaning of formal planning in public sector professional work. Curiously, this is an issue that remains under-investigated in the literature: the long debate on formal planning in strategy research devoted scant attention to its diffusion in the public sector, and public sector studies have scrutinized the introduction of other management tools in professional work, but very limitedly formal planning itself. In fact, little is known on the actual meaning of formal planning in public, professional services. This research is based upon a case of adoption of formal planning tools in a public hospital. Embracing a discourse analytical lens, it examines which formal planning discourse entered professional work, to what extent, and how professionals interpret it and engage with it in their practice. The analysis uncovers dynamics of social construction of meaning where, eventually, a formal planning discourse both shapes and is shaped by professional practice. In particular, it is found that formal planning rationality largely penetrated professional work, but not to the detriment of professional values. Morevover, formal planning ‘fails’ as a tool for rational decision making, but it takes up a knowledge work and a social value in professional work, as a tool for explicitation of action courses and for dialogue between otherwise more disconnected parts of the organization.

Essays in behavioral economics

In this work we conduct an experimental analysis on different behavioral models of economic choice. In particular, we analyze the role of overconfidence in shaping the beliefs of economics agents about the future path of their consumption or investment. We discuss the relevance of this bias in expectation formation both from a static and from a dynamic point of view and we analyze the effect of possible interventions aimed to achieve some policy goals. The methodology we follow is both theoretical and empirical. In particular, we make large use of controlled economic field experiments in order to test the predictions of the theoretical models we propose. In the second part of the thesis we discuss the role of cognition and personality in affecting economic preferences and choices. In this way we make a bridge between established psychological research and novel findings in economics. Finally, we conduct a field study on the role of incentives on education. We design different incentive schemes and we test, on randomized groups of students, their effectiveness in improving academic performance.

Untersuchungen des Keratinfilament-Turnovers in lebenden Zellen

Das Zytoskelett eukaryotischer Zellen besteht aus drei verschiedenen Protein-Netzwerken: den Aktinfilamenten, Mikrotubuli und Intermediärfilamenten. Intermediärfilamente wurden ursprünglich als statische Strukturen angesehen, die die mechanische Stabilisierung der Zellen übernehmen. In den letzten Jahren hat sich dieses Bild jedoch geändert: Intermediärfilament-Netzwerke sind hochdynamisch und unterliegen kontinuierlichen Veränderungen, welche durch Phosphorylierungen reguliert werden. Sie interagieren mit anderen Zytoskelett-Proteinen und greifen in die Regulation von Schlüsselsignalwegen, die Zellwachstum und Zellteilung sowie Apoptose und Stressantwort bestimmen, ein. Die Mechanismen der Filamentplastizität konnten bisher jedoch nicht vollständig aufgeklärt werden. So ist beispielsweise unklar, wo Auf- und Abbau der Filamente stattfindet und welche Faktoren an der Netzwerkmodulation beteiligt sind. Ziel meiner Arbeit war es, einen Beitrag zur Aufklärung dieser Mechanismen am Beispiel der epithelialen Keratin-Intermediärfilamente zu leisten. Mit Hilfe von mikroskopischen Zeitrafferaufnahmen von fluoreszenzmarkierten Zellklonen wurden Nukleationszentren in der Zellperipherie identifiziert, in denen Keratinfilamentvorläufer gebildet werden. Es handelt sich dabei um fokale Adhäsionskomplexe, die als Anheftungsstellen zwischen der extrazellulären Matrix und dem intrazellulären Aktinfilament-System dienen. Es konnte gezeigt werden, dass diese Filamentvorläufer-Entstehung für alle untersuchten Keratinisoformen gültig ist und in epitelialen als auch nicht-epithelialen Zelltypen abläuft. Knock-Down der Adhäsionskomponente Talin verhinderte die Keratinfilamentbildung. Modulation der fokalen Adhäsionskinase, die den Auf- und Abbau der Adhäsionskomplexe koordiniert, beeinflusste ebenso die Bildung der Keratinfilamentnetzwerke. Es konnte weiterhin beobachtet werden, dass die N-terminalen Isoformen IE und IF des Zytolinkers Plectin in fokalen Adhäsionen lokalisieren und damit möglicherweise an der Vernetzung von Keratinfilamentvorläufern, Zelladhäsionen und Aktinfilamenten beteiligt sind. Letztlich stellte sich heraus, dass die Bildung der Keratinfilamentvorläufer unabhängig von Proteintranslation ist. In den mikroskopischen Zeitrafferaufnahmen wurde im Anschluss an die Keratinfilamentbildung ein kontinuierlicher zentripetaler Transport der wachsenden Vorläuferpartikel beobachtet. An Hand von pharmakologischen Experimenten konnte gezeigt werden, dass dieser Transport Aktinfilament-abhängig ist. Zeitgleich kommt es zu Partikelfusion und Integration in das periphere Netzwerk, das sich weiterhin in Richtung auf das Zellzentrum bewegt. Mit Hilfe von Photoaktivierungsversuchen und Zellfusionsexperimenten konnte die Hypothese bestätigt werden, dass der Abbau der einwandernden Keratinfilamente in lösliche, rasch diffusible Zwischenstufen den kontinuierlichen peripheren Neuaufbau ermöglicht. Aus den Beobachtungen und bereits bekannten Ergebnissen wurde ein Modell des Keratin-Zyklus entwickelt, das die folgenden Stadien umfasst: Nukleation von Keratinfilamentvorläufern an fokalen Adhäsionen in der Zellperipherie, Elongation und Fusion der Keratinfilamentvorläufer bei zeitgleichem Aktinfilament-abhängigem zentripetalen Transport, Integration der Keratinfilamentvorläufer in das periphere Netzwerk, Bündelung der Filamente, Filamentabbau in lösliche Untereinheiten und Neubeginn des Zyklus in der Zellperipherie. Eine Störung dieses Zyklus liegt bei mutierten Keratinen vor, welche die Ursache von Blasen-bildenden Hauterkrankungen sind. In der vorliegenden Arbeit wurde am Beispiel von Keratin 6a-Mutanten, welche die Hauterkrankung Pachyonychia congenita verursachen, gezeigt, dass bei diesen Keratinen die Nukleation zwar im Bereich der Adhäsionskomplexe regelrecht abläuft, die anschließende Elongation und Netzwerkbildung aber gestört ist, so dass statt dessen kurzlebige, hyperphosphorylierte Granula entstehen. Der resultierende frustrane Keratin-Zyklus in der Zellperipherie ist stark beschleunigt und kann durch p38-Inhibierung gestoppt werden. Bei Proteasomeninhibierung wird der Zyklus in Richtung der Granulabildung verschoben. In dieser Arbeit wird erstmals das Keratin-Tretmühlen-Modell vorgestellt, das den regulierbaren Auf- und Abbau-Zyklus des Keratinnetzwerks beschreibt. Damit liegen testbare Hypothesen für die Aufklärung der Keratinfilament-Plastizität in physiologischen und pathologischen Situationen vor, die nach unseren ersten Ergebnissen auch von Relevanz für andere Intermediärfilamenttypen sind.

The behaviour of osteocyte dendritic processes in bone under cyclic load (il comportamento dei processi dendritici degli osteociti nel tessuto osseo sottoposti a carico ciclico)

Bone is continually being removed and replaced through the actions of basic multicellular units (BMU). This constant upkeep is necessary to remove microdamage formed naturally due to fatigue and thus maintain the integrity of the bone. The repair process in bone is targeted, meaning that a BMU travels directly to the site of damage and repairs it. It is still unclear how targeted remodelling is stimulated and directed but it is highly likely that osteocytes play a role. A number of theories have been advanced to explain the microcrack osteocyte interaction but no complete mechanism has been demonstrated. Osteocytes are connected to each other by dendritic processes. The “scissors model" proposed that the rupture of these processes where they cross microcracks signals the degree of damage and the urgency of the necessary repair. In its original form it was proposed that under applied compressive loading, microcrack faces will be pressed together and undergo relative shear movement. If this movement is greater than the width of an osteocyte process, then the process will be cut in a “scissors like" motion, releasing RANKL, a cytokine known to be essential in the formation of osteoclasts from pre-osteoclasts. The main aim of this thesis was to investigate this theoretical model with a specific focus on microscopy and finite element modelling. Previous studies had proved that cyclic stress was necessary for osteocyte process rupture to occur. This was a divergence from the original “scissors model" which had proposed that the cutting of cell material occurred in one single action. The present thesis is the first study to show fatigue failure in cellular processes spanning naturally occurring cracks and it's the first study to estimate the cyclic strain range and relate it to the number of cycles to failure, for any type of cell. Rupture due to shear movement was ruled out as microcrack closing never occurred, as a result of plastic deformation of the bone. Fatigue failure was found to occur due to cyclic tensile stress in the locality of the damage. The strain range necessary for osteocyte process rupture was quantified. It was found that the lower the process strain range the greater the number of cycles to cell process failure. FEM modelling allowed to predict stress in the vicinity of an osteocyte process and to analyse its interaction with the bone surrounding it: simulations revealed evident creep effects in bone during cyclic loading. This thesis confirms and dismisses aspects of the “scissors model". The observations support the model as a viable mechanism of microcrack detection by the osteocyte network, albeit in a slightly modified form where cyclic loading is necessary and the method of rupture is fatigue failure due to cyclic tensile motion. An in depth study was performed focusing on microscopy analysis of naturally occurring cracks in bone and FEM simulation analysis of an osteocyte process spanning a microcrack in bone under cyclic load.

Understanding the Combined Role of Internal and External Factors in New Service Development Process within the Professional Knowledge Intensive Business Service Firms

Our research asked the following main questions: how the characteristics of professionals service firms allow them to successfully innovate in exploiting through exploring by combining internal and external factors of innovation and how these ambidextrous organisations perceive these factors; and how do successful innovators in professional service firms use corporate entrepreneurship models in their new service development processes? With a goal to shed light on innovation in professional knowledge intensive business service firms’ (PKIBS), we concluded a qualitative analysis of ten globally acting law firms, providing business legal services. We analyse the internal and factors of innovation that are critical for PKIBS’ innovation. We suggest how these firms become ambidextrous in changing environment. Our findings show that this kind of firms has particular type of ambidexterity due to their specific characteristics. As PKIBS are very dependant on its human capital, governance structure, and the high expectations of their clients, their ambidexterity is structural, but also contextual at the same time. In addition, we suggest 3 types of corporate entrepreneurship models that international PKIBS use to enhance innovation in turbulent environments. We looked at how law firms going through turbulent environments were using corporate entrepreneurship activities as a part of their strategies to be more innovative. Using visual mapping methodology, we developed three types of innovation patterns in the law firms. We suggest that corporate entrepreneurship models depend on successful application of mainly three elements: who participates in corporate entrepreneurship initiatives; what are the formal processes that enhances these initiatives; and what are the policies applied to this type of behaviour.

Tissue engineering strategies for biomaterial-based anticancer drug delivery and enhancement of angiogenesis in tumor-resected bone

Patienten, die an Osteosarkom leiden werden derzeit mit intravenös applizierten krebstherapeutischen Mitteln nach Tumorresektion behandelt, was oftmals mit schweren Nebenwirkungen und einem verzögerten Knochenheilungsprozess einhergeht. Darüber hinaus treten vermehrt Rezidive aufgrund von verbleibenden neoplastischen Zellen an der Tumorresektionsstelle auf. Erfolgreiche Knochenregeneration und die Kontrolle von den im Gewebe verbleibenden Krebszellen stellt eine Herausforderung für das Tissue Engineering nach Knochenverlust durch Tumorentfernung dar. In dieser Hinsicht scheint der Einsatz von Hydroxyapatit als Knochenersatzmaterial in Kombination mit Cyclodextrin als Medikamententräger, vielversprechend. Chemotherapeutika können an Biomaterial gebunden und direkt am Tumorbett über einen längeren Zeitraum freigesetzt werden, um verbliebene neoplastische Zellen zu eliminieren. Lokal applizierte Chemotherapie hat diverse Vorteile, einschließlich der direkten zytotoxischen Auswirkung auf lokale Zellen, sowie die Reduzierung schwerer Nebenwirkungen. Diese Studie wurde durchgeführt, um die Funktionsfähigkeit eines solchen Arzneimittelabgabesystems zu bewerten und um Strategien im Bereich des Tissue Engineerings zu entwickeln, die den Knochenheilungsprozess und im speziellen die Vaskularisierung fördern sollen. Die Ergebnisse zeigen, dass nicht nur Krebszellen von der chemotherapeutischen Behandlung betroffen sind. Primäre Endothelzellen wie zum Beispiel HUVEC zeigten eine hohe Sensibilität Cisplatin und Doxorubicin gegenüber. Beide Medikamente lösten in HUVEC ein tumor-unterdrückendes Signal durch die Hochregulation von p53 und p21 aus. Zudem scheint Hypoxie einen krebstherapeutischen Einfluss zu haben, da die Behandlung sensitiver HUVEC mit Hypoxie die Zellen vor Zytotoxizität schützte. Der chemo-protektive Effekt schien deutlich weniger auf Krebszelllinien zu wirken. Diese Resultate könnten eine mögliche chemotherapeutische Strategie darstellen, um den Effekt eines zielgerichteten Medikamenteneinsatzes auf Krebszellen zu verbessern unter gleichzeitiger Schonung gesunder Zellen. Eine erfolgreiche Integration eines Systems, das Arzneimittel abgibt, kombiniert mit einem Biomaterial zur Stabilisierung und Regeneration, könnte gesunden Endothelzellen die Möglichkeit bieten zu proliferieren und Blutgefäße zu bilden, während verbleibende Krebszellen eliminiert werden. Da der Prozess der Knochengeweberemodellierung mit einer starken Beeinträchtigung der Lebensqualität des Patienten einhergeht, ist die Beschleunigung des postoperativen Heilungsprozesses eines der Ziele des Tissue Engineerings. Die Bildung von Blutgefäßen ist unabdingbar für eine erfolgreiche Integration eines Knochentransplantats in das Gewebe. Daher ist ein umfangreich ausgebildetes Blutgefäßsystem für einen verbesserten Heilungsprozess während der klinischen Anwendung wünschenswert. Frühere Experimente zeigen, dass sich die Anwendung von Ko-Kulturen aus humanen primären Osteoblasten (pOB) und humanen outgrowth endothelial cells (OEC) im Hinblick auf die Bildung stabiler gefäßähnlicher Strukturen in vitro, die auch effizient in das mikrovaskuläre System in vivo integriert werden konnten, als erfolgreich erweisen. Dieser Ansatz könnte genutzt werden, um prä-vaskularisierte Konstrukte herzustellen, die den Knochenheilungsprozess nach der Implantation fördern. Zusätzlich repräsentiert das Ko-Kultursystem ein exzellentes in vitro Model, um Faktoren, welche stark in den Prozess der Knochenheilung und Angiogenese eingebunden sind, zu identifizieren und zu analysieren. Es ist bekannt, dass Makrophagen eine maßgebliche Rolle in der inflammatorisch-induzierten Angiogenese spielen. In diesem Zusammenhang hebt diese Studie den positiven Einfluss THP-1 abgeleiteter Makrophagen in Ko-Kultur mit pOB und OEC hervor. Die Ergebnisse zeigten, dass die Anwendung von Makrophagen als inflammatorischer Stimulus im bereits etablierten Ko-Kultursystem zu einer pro-angiogenen Aktivierung der OEC führte, was in einer signifikant erhöhten Bildung blutgefäßähnlicher Strukturen in vitro resultierte. Außerdem zeigte die Analyse von Faktoren, die in der durch Entzündung hervorgerufenen Angiogenese eine wichtige Rolle spielen, eine deutliche Hochregulation von VEGF, inflammatorischer Zytokine und Adhäsionsmoleküle, die letztlich zu einer verstärkten Vaskularisierung beitragen. Diese Resultate werden dem Einfluss von Makrophagen zugeschrieben und könnten zukünftig im Tissue Engineering eingesetzt werden, um den Heilungsprozess zu beschleunigen und damit die klinische Situation von Patienten zu verbessern. Darüber hinaus könnte die Kombination der auf Ko-Kulturen basierenden Ansätze für das Knochen Tissue Engineering mit einem biomaterial-basierenden Arzneimittelabgabesystem zum klinischen Einsatz kommen, der die Eliminierung verbliebener Krebszellen mit der Förderung der Knochenregeneration verbindet.

Analysis of new bone formation induced by periosteal distraction in a rat calvarium model

Background: A controlled, gradual distraction of the periosteum is expected to result in the formation of new bone. Purpose: This study was designed to estimate the possibility of new bone formation by periosteal distraction in a rat calvarium model. Material and Methods: Sixteen animals were subjected to a 7-day latency period and distraction rate at 0.4 mm/24 hours for 10 days. Two experimental groups with seven rats each were killed at 10 and 20 days of consolidation period and analyzed by means of microcomputed tomography, histologically and histomorphometry. Results: In the central regions underneath the disk device, signs of both bone apposition and bone resorption were observed. Peripheral to the disc, new bone was consistently observed. This new bone was up to two and three times thicker than the original bone after a 10- and 20-day consolidation period, respectively. Signs of ongoing woven bone formation indicated that the stimulus for new bone formation was still present. There were no statistically significant differences regarding bone density, bone volume, and total bone height between the two groups. Conclusion: The periosteal distraction model in the rat calvarium can stimulate the formation of considerable amounts of new bone.

Frontal areas contribute to reduced global coordination of resting-state gamma activities in drug-naïve patients with schizophrenia

Schizophrenia has been postulated to involve impaired neuronal cooperation in large-scale neural networks, including cortico-cortical circuitry. Alterations in gamma band oscillations have attracted a great deal of interest as they appear to represent a pathophysiological process of cortical dysfunction in schizophrenia. Gamma band oscillations reflect local cortical activities, and the synchronization of these activities among spatially distributed cortical areas has been suggested to play a central role in the formation of networks. To assess global coordination across spatially distributed brain regions, Omega complexity (OC) in multichannel EEG was proposed. Using OC, we investigated global coordination of resting-state EEG activities in both gamma (30–50 Hz) and below-gamma (1.5–30 Hz) bands in drug-naïve patients with schizophrenia and investigated the effects of neuroleptic treatment. We found that gamma band OC was significantly higher in drug-naïve patients with schizophrenia compared to control subjects and that a right frontal electrode (F3) contributed significantly to the higher OC. After neuroleptic treatment, reductions in the contribution of frontal electrodes to global OC in both bands correlated with the improvement of schizophrenia symptomatology. The present study suggests that frontal brain processes in schizophrenia were less coordinated with activity in the remaining brain. In addition, beneficial effects of neuroleptic treatment were accompanied by improvement of brain coordination predominantly due to changes in frontal regions. Our study provides new evidence of improper intrinsic brain integration in schizophrenia by investigating the resting-state gamma band activity.

Biogeochemical processes in a clay formation in situ experiment: Part F - Reactive transport modelling

Reactive transport modelling was used to simulate solute transport, thermodynamic reactions, ion exchange and biodegradation in the Porewater Chemistry (PC) experiment at the Mont Terri Rock Laboratory. Simulations show that the most important chemical processes controlling the fluid composition within the borehole and the surrounding formation during the experiment are ion exchange, biodegradation and dissolution/precipitation reactions involving pyrite and carbonate minerals. In contrast, thermodynamic mineral dissolution/precipitation reactions involving alumo-silicate minerals have little impact on the fluid composition on the time-scale of the experiment. With the accurate description of the initial chemical condition in the formation in combination with kinetic formulations describing the different stages of bacterial activities, it has been possible to reproduce the evolution of important system parameters, such as the pH, redox potential, total organic C. dissolved inorganic C and SO(4) concentration. Leaching of glycerol from the pH-electrode may be the primary source of organic material that initiated bacterial growth, which caused the chemical perturbation in the borehole. Results from these simulations are consistent with data from the over-coring and demonstrate that the Opalinus Clay has a high buffering capacity in terms of chemical perturbations caused by bacterial activity. This buffering capacity can be attributed to the carbonate system as well as to the reactivity of clay surfaces.