Produzione di virus sintetici per lo studio dei meccanismi di interazione coinvolti nell'induzione di resistenza

Beet soil-borne mosaic virus (BSBMV) and Beet necrotic yellow vein virus (BNYVV) are members of Benyvirus genus. BSBMV has been reported only in the United States while BNYVV has a worldwide distribution. Both viruses are vectored by Polymyxa betae, possess similar host ranges, particles number and morphology. Both viruses are not serologically related but have similar genomic organizations. Field isolates consist of four RNA species but some BNYVV isolates contain a fifth RNA. RNAs 1 and 2 are essential for infection and replication while RNAs 3 and 4 play important roles on plant and vector interactions, respectively. Nucleotide and amino acid analyses revealed BSBMV and BNYVV are different enough to be classified in two different species. Additionally in BNYVV/BSBMV mixed infections, a competition was previous described in sugar beet, where BNYVV infection reduces BSBMV accumulation in both susceptible and resistant cultivars. Considering all this observations we hypothesized that BNYVV and BSBMV crossed study, exploiting their similarities and divergences, can improve investigation of molecular interactions between sugar beets and Benyviruses. The main achievement of our research is the production of a cDNA biologically active clones collection of BNYVV and BSBMV RNAs, from which synthetic copies of both Benyviruses can be transcribed. Moreover, through recombination experiments we demonstrated, for the first time, the BNYVV RNA 1 and 2 capability to trans-replicate and encapsidate BSBMV RNA 3 and 4, either the BSBMV RNA 1 and 2 capability to replicate BNYVV RNA2 in planta. We also demonstrated that BSBMV RNA3 support long-distance movement of BNYVV RNA 1 and 2 in B. macrocarpa and that 85 foreign sequence as p29HA, GFP and RFP, are successfully expressed, in C. quinoa, by BSBMV RNA3 based replicon (RepIII) also produced by our research. These results confirm the close correlation among the two viruses. Interestingly, the symptoms induced by BSBMV RNA-3 on C. quinoa leaves are more similar to necrotic local lesions caused by BNYVV RNA-5 p26 than to strongly chlorotic local lesions or yellow spot induced by BNYVV RNA- 3 encoded p25. As previous reported BSBMV p29 share 23% of amino acid sequence identity with BNYVV p25 but identity increase to 43% when compared with sequence of BNYVV RNA-5 p26. Based on our results the essential sequence (Core region) for the longdistance movement of BSBMV and BNYVV in B. macrocarpa, is not only carried by RNA3s species but other regions, perhaps located on the RNA 1 and 2, could play a fundamental role in this matter. Finally a chimeric RNA, composed by the 5’ region of RNA4 and 3’ region of RNA3 of BSBMV, has been produced after 21 serial mechanically inoculation of wild type BSBMV on C. quinoa plants. Chimera seems unable to express any protein, but it is replicated and transcript in planta. It could represent an important tool to study the interactions between Benyvirus and plant host. In conclusion different tools, comprising a method to study synthetic viruses under natural conditions of inoculum through P. Betae, have been produced and new knowledge are been acquired that will allow to perform future investigation of the molecular interactions between sugar beets and Benyviruses.

Precision horticulture: application on apple orchards

Precision horticulture and spatial analysis applied to orchards are a growing and evolving part of precision agriculture technology. The aim of this discipline is to reduce production costs by monitoring and analysing orchard-derived information to improve crop performance in an environmentally sound manner. Georeferencing and geostatistical analysis coupled to point-specific data mining allow to devise and implement management decisions tailored within the single orchard. Potential applications range from the opportunity to verify in real time along the season the effectiveness of cultural practices to achieve the production targets in terms of fruit size, number, yield and, in a near future, fruit quality traits. These data will impact not only the pre-harvest but their effect will extend to the post-harvest sector of the fruit chain. Chapter 1 provides an updated overview on precision horticulture , while in Chapter 2 a preliminary spatial statistic analysis of the variability in apple orchards is provided before and after manual thinning; an interpretation of this variability and how it can be managed to maximize orchard performance is offered. Then in Chapter 3 a stratification of spatial data into management classes to interpret and manage spatial variation on the orchard is undertaken. An inverse model approach is also applied to verify whether the crop production explains environmental variation. In Chapter 4 an integration of the techniques adopted before is presented. A new key for reading the information gathered within the field is offered. The overall goal of this Dissertation was to probe into the feasibility, the desirability and the effectiveness of a precision approach to fruit growing, following the lines of other areas of agriculture that already adopt this management tool. As existing applications of precision horticulture already had shown, crop specificity is an important factor to be accounted for. This work focused on apple because of its importance in the area where the work was carried out, and worldwide.

Methodological improvement of 3D fluoroscopic analysis for the robust quantification of 3D kinematics of human joints

3D video-fluoroscopy is an accurate but cumbersome technique to estimate natural or prosthetic human joint kinematics. This dissertation proposes innovative methodologies to improve the 3D fluoroscopic analysis reliability and usability. Being based on direct radiographic imaging of the joint, and avoiding soft tissue artefact that limits the accuracy of skin marker based techniques, the fluoroscopic analysis has a potential accuracy of the order of mm/deg or better. It can provide fundamental informations for clinical and methodological applications, but, notwithstanding the number of methodological protocols proposed in the literature, time consuming user interaction is exploited to obtain consistent results. The user-dependency prevented a reliable quantification of the actual accuracy and precision of the methods, and, consequently, slowed down the translation to the clinical practice. The objective of the present work was to speed up this process introducing methodological improvements in the analysis. In the thesis, the fluoroscopic analysis was characterized in depth, in order to evaluate its pros and cons, and to provide reliable solutions to overcome its limitations. To this aim, an analytical approach was followed. The major sources of error were isolated with in-silico preliminary studies as: (a) geometric distortion and calibration errors, (b) 2D images and 3D models resolutions, (c) incorrect contour extraction, (d) bone model symmetries, (e) optimization algorithm limitations, (f) user errors. The effect of each criticality was quantified, and verified with an in-vivo preliminary study on the elbow joint. The dominant source of error was identified in the limited extent of the convergence domain for the local optimization algorithms, which forced the user to manually specify the starting pose for the estimating process. To solve this problem, two different approaches were followed: to increase the optimal pose convergence basin, the local approach used sequential alignments of the 6 degrees of freedom in order of sensitivity, or a geometrical feature-based estimation of the initial conditions for the optimization; the global approach used an unsupervised memetic algorithm to optimally explore the search domain. The performances of the technique were evaluated with a series of in-silico studies and validated in-vitro with a phantom based comparison with a radiostereometric gold-standard. The accuracy of the method is joint-dependent, and for the intact knee joint, the new unsupervised algorithm guaranteed a maximum error lower than 0.5 mm for in-plane translations, 10 mm for out-of-plane translation, and of 3 deg for rotations in a mono-planar setup; and lower than 0.5 mm for translations and 1 deg for rotations in a bi-planar setups. The bi-planar setup is best suited when accurate results are needed, such as for methodological research studies. The mono-planar analysis may be enough for clinical application when the analysis time and cost may be an issue. A further reduction of the user interaction was obtained for prosthetic joints kinematics. A mixed region-growing and level-set segmentation method was proposed and halved the analysis time, delegating the computational burden to the machine. In-silico and in-vivo studies demonstrated that the reliability of the new semiautomatic method was comparable to a user defined manual gold-standard. The improved fluoroscopic analysis was finally applied to a first in-vivo methodological study on the foot kinematics. Preliminary evaluations showed that the presented methodology represents a feasible gold-standard for the validation of skin marker based foot kinematics protocols.

Cambiamenti dello scenario epidemiologico e clinico dell'epatocarcinoma in Italia nell'ultimo ventennio

Background & Aims: This study investigates whether the aetiologic changes in liver disease and the improved management of hepatocellular carcinoma (HCC) have modified the clinical scenario of this tumour over the last 20 years in Italy. Methods: Retrospective study based on the analysis of the ITA.LI.CA (Italian Liver Cancer) database including 3027 HCC patients managed in 11 centres. Patients were divided into 3 groups according to the period of HCC diagnosis: 1987–1996 (year of the ‘‘Milano criteria’’ publication), 1997–2001 (year of release of the EASL guidelines for HCC), and 2002–2008. Results: The significant changes were: (1) progressive patient ageing; (2) increasing prevalence of HCV infection until 2001, with a subsequent decrease, when the alcoholic aetiology increased; (3) liver function improvement, until 2001; (4) increasing ‘‘incidental’’ at the expense of ‘‘symptomatic’’ diagnoses, until 2001; (5) unchanged prevalence of tumours diagnosed during surveillance (around 50%), with an increasing use of the 6- month schedule; (6) favourable HCC ‘‘stage migration’’, until 2001; (7) increasing use of percutaneous ablation; (8) improving survival, until 2001. Conclusions: Over the last 20 years, several aetiologic and clinical features regarding HCC have changed. The survival improvement observed until 2001 was due to an increasing number of tumours diagnosed in early stages and in a background of compensated cirrhosis, and a growing and better use of locoregional treatments. However, the prevalence of early cancers and survival did not increase further in the last years, a result inciting national policies aimed at implementing surveillance programmes for at risk patients.

Le aziende vitivinicole: da “machines à produire” a elementi di attrazione nel paesaggio. Proposte e criteri progettuali per l’efficienza funzionale e la qualità architettonica ed ambientale delle strutture ed infrastrutture della trasformazione enologica.

La ricerca ha per oggetto la messa a punto e applicazione di un approccio metaprogettuale finalizzato alla definizione di criteri di qualità architettonica e paesaggistica nella progettazione di aziende vitivinicole medio-piccole, che effettuano la trasformazione della materia prima, prevalentemente di propria produzione. L’analisi della filiera vitivinicola, della letteratura scientifica, della normativa di settore, di esempi di “architetture del vino eccellenti” hanno esplicitato come prevalentemente vengano indagate cantine industriali ed aspetti connessi con l'innovazione tecnologica delle attrezzature. Soluzioni costruttive e tecnologiche finalizzate alla qualità architettonica ed ambientale, attuali dinamiche riguardanti il turismo enogastronomico, nuove funzionalità aziendali, problematiche legate alla sostenibilità dell’intervento risultano ancora poco esplorate, specialmente con riferimento a piccole e medie aziende vitivinicole. Assunto a riferimento il territorio ed il sistema costruito del Nuovo Circondario Imolese (areale rappresentativo per vocazione ed espressione produttiva del comparto vitivinicolo emiliano-romagnolo) è stato identificato un campione di aziende con produzioni annue non superiori ai 5000 hl. Le analisi svolte sul campione hanno permesso di determinare: modalità di aggregazione funzionale degli spazi costruiti, relazioni esistenti con il paesaggio, aspetti distributivi e materico-costruttivi, dimensioni di massima dei locali funzionali alla produzione. Il caso studio relativo alla riqualificazione di un’azienda rappresentativa del comparto è stato utilizzato per la messa a punto e sperimentazione di criteri di progettazione guidati da valutazioni relative alle prestazioni energetiche, alla qualità architettonica e alla sostenibilità ambientale, economica e paesaggistica. L'analisi costi-benefici (pur non considerando le ricadute positive in termini di benessere degli occupanti ed il guadagno della collettività in termini di danni collegati all’inquinamento che vengono evitati in architetture progettate per garantire qualità ambientale interna ed efficienza energetica) ha esplicitato il ritorno in pochi anni dell’investimento proposto, nonostante gli ancora elevati costi di materiali di qualità e dei componenti per il corretto controllo climatico delle costruzioni.

Expression profiling und rationale Expressionsoptimierung am Beispiel eines prokaryontischen Wirt-,Vektor-Systems

Durch globale Expressionsprofil-Analysen auf Transkriptom-, Proteom- oder Metabolom-Ebene können biotechnologische Produktionsprozesse besser verstanden und die Erkenntnisse für die zielgerichtete, rationale Optimierung von Expressionssystemen genutzt werden. In der vorliegenden Arbeit wurde die Überexpression einer Glukose-Dehydrogenase (EC 1.1.5.2), die von der Roche Diagnostics GmbH für die diagnostische Anwendung optimiert worden war, in Escherichia coli untersucht. Die Enzymvariante unterscheidet sich in sieben ihrer 455 Aminosäuren vom Wildtyp-Enzym und wird im sonst isogenen Wirt-/Vektor-System in signifikant geringeren Mengen (Faktor 5) gebildet. Das prokaryontische Expressionssystem wurde auf Proteom-Ebene charakterisiert. Die 2-dimensionale differenzielle Gelelektrophorese (DIGE) wurde zuvor unter statistischen Aspekten untersucht. Unter Berücksichtigung von technischen und biologischen Variationen, falsch-positiven (α-) und falsch-negativen (β-) Fehlern sowie einem daraus abgeleiteten Versuchsdesign konnten Expressionsunterschiede als signifikant quantifiziert werden, wenn sie um den Faktor ≥ 1,4 differierten. Durch eine Hauptkomponenten-Analyse wurde gezeigt, dass die DIGE-Technologie für die Expressionsprofil-Analyse des Modellsystems geeignet ist. Der Expressionsstamm für die Enzymvariante zeichnete sich durch eine höhere Variabilität an Enzymen für den Zuckerabbau und die Nukleinsäure-Synthese aus. Im Expressionssystem für das Wildtyp-Enzym wurde eine unerwartet erhöhte Plasmidkopienzahl nachgewiesen. Als potenzieller Engpass in der Expression der rekombinanten Glukose-Dehydrogenase wurde die Löslichkeitsvermittlung identifiziert. Im Expressionsstamm für das Wildtyp-Enzym wurden viele Proteine für die Biogenese der äußeren Membran verstärkt exprimiert. Als Folge dessen wurde ein sog. envelope stress ausgelöst und die Zellen gingen in die stationäre Wuchsphase über. Die Ergebnisse der Proteomanalyse wurden weiterführend dazu genutzt, die Produktionsleistung für die Enzymvariante zu verbessern. Durch den Austausch des Replikationsursprungs im Expressionsvektor wurde die Plasmidkopienzahl erhöht und die zelluläre Expressionsleistung für die diagnostisch interessantere Enzymvariante um Faktor 7 - 9 gesteigert. Um die Löslichkeitsvermittlung während der Expression zu verbessern, wurde die Plasmidkopienzahl gesenkt und die Coexpression von Chaperonen initiiert. Die Ausbeuten aktiver Glukose-Dehydrogenase wurden durch die Renaturierung inaktiven Produkts aus dem optimierten Expressionssystem insgesamt um einen Faktor von 4,5 erhöht. Somit führte im Rahmen dieser Arbeit eine proteombasierte Expressionsprofil-Analyse zur zielgerichteten, rationalen Expressionsoptimierung eines prokaryontischen Modellsystems.

Optimization of bioenergy solutions at different farm scales

RAF is a bio-energetic descriptive model integrates with MAD model to support Integrated Farm Management. RAF model aimed to enhancing economical, social and environmental sustainability of farm production in terms of energy via convert energy crops and animal manure to biogas and digestate (bio-fertilizers) by anaerobic digestion technologies, growing and breeding practices. The user defines farm structure in terms of present crops, livestock and market prices and RAF model investigates the possibilities of establish on-farm biogas system (different anaerobic digestion technologies proposed for different scales of farms in terms of energy requirements) according to budget and sustainability constraints to reduce the dependence on fossil fuels. The objective function of RAF (Z) is optimizing the total net income of farm (maximizing income and minimizing costs) for whole period which is considered by the analysis. The main results of this study refers to the possibility of enhancing the exploitation of the available Italian potentials of biogas production from on-farm production of energy crops and livestock manure feedstock by using the developed mathematical model RAF integrates with MAD to presents reliable reconcile between farm size, farm structure and on-farm biogas systems technologies applied to support selection, applying and operating of appropriate biogas technology at any farm under Italian conditions.

Modellazione edilizia e simulazione energetica per le analisi delle prestazioni e la progettazione integrata delle cantine aziendali

Il presente studio si colloca nell’ambito di una ricerca il cui obiettivo è la formulazione di criteri progettuali finalizzati alla ottimizzazione delle prestazioni energetiche delle cantine di aziende vitivinicole con dimensioni produttive medio-piccole. Nello specifico la ricerca si pone l’obiettivo di individuare degli indicatori che possano valutare l’influenza che le principali variabili progettuali hanno sul fabbisogno energetico dell’edificio e sull’andamento delle temperature all’interno dei locali di conservazione ed invecchiamento del vino. Tali indicatori forniscono informazioni sulla prestazione energetica dell’edificio e sull’idoneità dei locali non climatizzati finalizzata alla conservazione del vino Essendo la progettazione una complessa attività multidisciplinare, la ricerca ha previsto l’ideazione di un programma di calcolo in grado di gestire ed elaborare dati provenienti da diversi ambiti (ingegneristici, architettonici, delle produzioni agroindustriali, ecc.), e di restituire risultati sintetici attraverso indicatori allo scopo individuati. Il programma è stato applicato su un caso-studio aziendale rappresentativo del settore produttivo. Sono stati vagliati gli effetti di due modalità di vendemmia e di quattro soluzioni architettoniche differenti. Le soluzioni edilizie derivano dalla combinazione di diversi isolamenti termici e dalla presenza o meno di locali interrati. Per le analisi sul caso-studio ci si è avvalsi di simulazioni energetiche in regime dinamico, supportate e validate da campagne di monitoraggio termico e meteorologico all’interno dell’azienda oggetto di studio. I risultati ottenuti hanno evidenziato come il programma di calcolo concepito nell’ambito di questo studio individui le criticità dell’edificio in termini energetici e di “benessere termico” del vino e consenta una iterativa revisione delle variabili progettuale indagate. Esso quindi risulta essere uno strumento informatizzato di valutazione a supporto della progettazione, finalizzato ad una ottimizzazione del processo progettuale in grado di coniugare, in maniera integrata, gli obiettivi della qualità del prodotto, della efficienza produttiva e della sostenibilità economica ed ambientale.

Miglioramento delle performance termiche e energetiche degli edifici rurali: valutazione del retrofit di locali per la conservazione del vino su casi studio

Il presente studio si colloca all’interno di una ricerca più ampia volta alla definizione di criteri progettuali finalizzati all’ottimizzazione delle prestazioni energetiche delle cantine di aziende vitivinicole, di dimensioni produttive medio - piccole. Nello specifico la ricerca riguarda la riqualificazione di fabbricati rurali esistenti di modeste dimensioni, da convertire a magazzini per la conservazione del vino in bottiglia. Lo studio si pone come obiettivo la definizione di criteri di analisi per la valutazione di interventi di retrofit di tali fabbricati, volto sia al miglioramento delle prestazioni energetiche dell’involucro edilizio, sia alla riduzione del fabbisogno energetico legato al funzionamento di eventuali impianti di controllo termico. La ricerca è stata condotta mediante l’utilizzo del software di simulazione termica Energy Plus, per ottenere i valori simulati di temperatura interna relativi ai diversi scenari migliorativi ipotizzati, e mediante la successiva definizione di indicatori che esplicitino l’influenza delle principali variabili progettuali sull’andamento delle temperature interne dei locali di conservazione e sul fabbisogno energetico del fabbricato necessario a garantire l’intervallo di temperatura di comfort del vino. Tra tutti gli interventi possibili per il miglioramento della prestazione energetica degli edifici, quelli analizzati in questo studio prevedono l’aggiunta di un isolamento a cappotto delle pareti esterne, l’isolamento della copertura e l’aggiunta di una struttura ombreggiante vegetale esterna. I risultati ottenuti danno una prima indicazione sugli interventi più efficaci in termini di miglioramento energetico e mettono in luce l’utilità del criterio proposto nell’evidenziare le criticità degli interventi migliorativi ipotizzati. Il metodo definito nella presente ricerca risulta quindi un valido strumento di valutazione a supporto della progettazione degli interventi di retrofit dei fabbricati rurali da convertire a magazzini per la conservazione del vino.

Intralesional bovine papillomavirus DNA loads reflect severity of equine sarcoid disease

REASONS FOR PERFORMING STUDY: Sarcoids are nonmetastasising, yet locally aggressive skin tumours that constitute the most frequent neoplasm in equids. Infection by bovine papillomaviruses types 1 and 2 (BPV-1, BPV-2) has been recognised as major causative factor in sarcoid pathogenesis, but a possible correlation of intralesional virus load with disease severity has not been established thus far. HYPOTHESIS: Given the pathogenic role of BPV-1 and BPV-2 in sarcoid disease, we suggest that intralesional viral DNA concentration may reflect the degree of affection. METHODS: Severity of disease was addressed by recording the tumour growth kinetics, lesion number and tumour type for 37 sarcoid-bearing horses and one donkey. Viral load was estimated via quantitative real-time PCR (qPCR) of the E2, E5, L1 and L2 genes from the BPV-1/-2 genome for one randomly selected lesion per horse and correlated with disease severity. RESULTS: Quantitative PCR against E2 identified viral DNA concentrations ranging from 0-556 copies/tumour cell. Of 16 horses affected by quiescent, slowly growing single tumours or multiple mild-type lesions, 15 showed a viral load up to 1.4 copies per cell. In stark contrast, all equids (22/22) bearing rapidly growing and/or multiple aggressive sarcoids had a viral load between 3 and 569 copies per cell. Consistent results were obtained with qPCR against E5, L1 and L2. CONCLUSIONS: While tumours of the same clinical type carried variable virus load, confirming that viral titre does not determine clinical appearance, we identified a highly significant correlation between intralesional viral load and disease severity. POTENTIAL RELEVANCE: The rapid determination of BPV viral load will give a reliable marker for disease severity and may also be considered when establishing a therapeutic strategy.

Social Policy Facing the Problems of Youth Employment

Rumiana Stoilova (Bulgaria). Social Policy Facing the Problems of Youth Employment. Ms. Stoilova is a researcher in the Institute of Sociology in Sofia and worked on this project from October 1996 to September 1998. This project involved collecting both statistical and empirical data on the state of youth employment in Bulgaria, which was then compared with similar data from other European countries. One significant aspect was the parallel investigation of employment and unemployment, which took as a premise the continuity of professional experience where unemployment is just a temporary condition caused by external and internal factors. These need to be studied and changed on a systematic basis so as to create a more favourable market situation and to improve individuals' resources for improving their market opportunities. A second important aspect of the project was an analysis of the various entities active on the labour market, including government and private institutions, associations of unemployed persons, of employers or of trade unions, all with their specific legal powers and interests, and of the problems in communication between these. The major trends in youth unemployment during the period studied include a high proportion of the registered unemployed who are not eligible for social assistance, a lengthening of the average period of unemployment, an increase in the percentage of people who are unemployed for the first time and an increasing percentage of these who are not eligible for assistance, particularly among newly registered young people. At the same time the percentage of those for who work has been found is rising and during the last three years an increasing number of the unemployed have started some independent economic activity. Regional differences are also considerable and in the case of the Haskovo region represent a danger of losing the youngest generation, with resulting negative demographic effects. One major weakness of the existing institutional structure is the large scale of the black labour market, with clear negative implications for the young people drawn into it. The role of non-governmental organisations in providing support and information for the unemployed is growing and the government has recently introduced special preferences for organisations offering jobs to unemployed persons. Social policy in the labour market has however been largely restricted to passive measures, mostly because of the risk that poverty poses to people continuously excluded from the labour market. Among the active measures taken, well over half are concerned with providing jobs for the unemployed and there are very limited programmes for providing or improving qualifications. The nature of youth employment in Bulgaria can be seen in the influence of sustained structures (generation) and institutions (family and school). Ms. Stoilova studied the situation of the modern generation through a series of profiles, mostly those of continuously unemployed and self-employed persons, but also distinguishing between students and the unemployed, and between high school and university students. The different categories of young people were studied in separate mini-studies and the survey was carried out in five town in order to gather objective and subjective information on the state of the labour market in the different regions. She conducted interviews with several hundred young people covering questions of family background, career plans, attitudes to the labour situation and government measures to deal with it, and such questions as independence, mobility, attitude to work, etc. The interviews with young people unemployed for a long period of time show the risk involved in starting work and its link with dynamics of economic development. Their approval of structural reforms, of the financial restrictions connected with the introduction of a currency board and the inevitability of unemployment was largely declarative. The findings indicate that the continuously unemployed need practical knowledge and skills to "translate" the macroeconomic realities in concrete alternatives of individual work and initiative. The unemployed experience their exclusion from the labour market not only as a professional problem but also as an existential threat, of poverty, forced mobility and dependence on their parents' generation. The exclusion from the market of goods and services means more than just exercising restraint in their consumption, as it places restrictions on their personal development. Ms. Stoilova suggests that more efficient ways of providing financial aid and mobilisation are needed to counteract the social disintegration and marginalisation of the continuously unemployed. In measuring the speed of reform, university students took both employment opportunities and the implementation of the meritocratic principle in employment into account. When offered a hypothetical choice between a well-paid job and work in one's own profession, 62% would prefer opt for the well-paid job and for working for a company that offered career opportunities rather than employment in a family or own company. While most see the information gained during their studies as useful and interesting, relatively few see their education as competitive on a wider level and many were pessimistic about employment opportunities based on their qualifications. Very similar attitudes were found among high school students, with differences being due rather to family and personal situations. The unemployed, on the other hand, placed greater emphasis on possibilities of gaining or improving qualifications on a job and for the opportunities it would offer for personal contacts. High school students tend to attribute more significance to opportunities for personal accomplishment. A significant difference that five times fewer high school students were willing to work for state-owned companies, and many fewer expected to find permanent employment or to find a job in the area where they lived, Within the family situation, actual support for children seems to be higher than the feelings of confidence expressed in interviews. The attitudes of the families towards past experience seems to be linked with their ability to cope with the difficulties of the present, with those families which show an optimistic and active attitude towards the future having a greater respect for parents experience and tolerance in communication between parents and children.

Transdisciplinary co-production of knowledge in the development of organic agriculture in Switzerland

The present study analyses transdisciplinary co-production of knowledge in the development of organic farming in Switzerland by using Fleck's theory of thought styles and thought collectives. Three different phases can be identified throughout the historical development. The initial phase lasting from the beginning of the 1920s to the early 1970s contains numerous characteristics of diverse well-established definitions and concepts of transdisciplinarity and represents a successful transdisciplinary process, which has not been perceived as such in the past and present scientific discussion. The second and third phases show an increasing segregation of thought collectives, caused by internal changes such as the establishment of specialised research institutions and external processes like agriculture policy and market development. These developments led to a decreasing degree of transdisciplinarity. We observe an ambiguous trend: the continuously growing and today well-established positive societal recognition of an initially rather little accepted newcomer movement is associated with the gradual loss of its very valuable forms of knowledge co-production and the related philosophical background. In order to maintain the various forms of transdisciplinary co-production of knowledge, one has to reflect not only their results or outcome but also the whole cooperation process, which has led to these results. The understanding of the historical development and characteristic features of knowledge co-production as presented in this study will help to reinforce transdisciplinary research in organic agriculture and research on transdisciplinarity in general.

Dissection of floral pollination syndromes in petunia

Animal-mediated pollination is essential in the reproductive biology of many flowering plants and tends to be associated with pollination syndromes, sets of floral traits that are adapted to particular groups of pollinators. The complexity and functional convergence of various traits within pollination syndromes are outstanding examples of biological adaptation, raising questions about their mechanisms and origins. In the genus Petunia, complex pollination syndromes are found for nocturnal hawkmoths (P. axillaris) and diurnal bees (P. integrifolia), with characteristic differences in petal color, corolla shape, reproductive organ morphology, nectar quantity, nectar quality, and fragrance. We dissected the Petunia syndromes into their most important phenotypic and genetic components. They appear to include several distinct differences, such as cell-growth and cell-division patterns in the basal third of the petals, elongation of the ventral stamens, nectar secretion and nectar sugar metabolism, and enzymatic differentiation in the phenylpropanoid pathway. In backcross-inbred lines of species-derived chromosome segments in a transposon tagging strain of P. hybrida, one to five quantitative trait loci were identified for each syndrome component. Two loci for stamen elongation and nectar volume were confirmed in introgression lines and showed large allelic differences. The combined data provide a framework for a detailed understanding of floral syndromes from their developmental and molecular basis to their impact on animal behavior. With its molecular genetic tools, this Petunia system provides a novel venue for a pattern of adaptive radiation that is among the most characteristic of flowering plants.

Foreign Direct Investment in Agriculture: Land Grab or Food Security Improvement?

“Large-scale acquisition of land by foreign investors” is the correct term for a process where the verdict of guilt is often quicker than the examination. But is there something really new about land grab except in its extent? In comparison with colonial and post-colonial plantation operations, should foreign investors today behave differently? We generally accept coffee and banana exports as pro-growth and pro-development, just as for cars, beef and insurance. What then is wrong with an investment contract allowing the holder to buy a farm and to export wheat to Saudi Arabia, or soybeans and maize as cattle feed to Korea, or to plant and process sugar cane and palm oil into ethanol for Europe and China? Assuming their land acquisition was legal, should foreigners respect more than investment contracts and national legislation? And why would they not take advantage of the legal protection offered by international investment law and treaties, not to speak of concessional finance, infrastructure and technical cooperation by a development bank, or the tax holidays offered by the host state? Remember Milton Friedman’s often-quoted quip: “The business of business is business!” And why would the governments signing those contracts not know whether and which foreign investment projects are best for their country, and how to attract them? This chapter tries to show that land grab, where it occurs, is not only yet another symptom of regulatory failures at the national level and a lack of corporate social responsibility by certain private actors. National governance is clearly the most important factor. Nonetheless, I submit that there is an international dimension involving investor home states in various capacities. The implication is that land grab is not solely a question whether a particular investment contract is legal or not. This chapter deals with legal issues which seem to have largely escaped the attention of both human rights lawyers and, especially, of investment lawyers. I address this fragmentation between different legal disciplines, rules, and policies, by asking two basic questions: (i) Do governments and parliaments in investor home countries have any responsibility in respect of the behaviour of their investors abroad? (ii) What should they and international regulators do, if anything?

Favourable mid-term outcome after heart transplantation for late Fontan failure.

OBJECTIVES Fontan failure (FF) represents a growing and challenging indication for paediatric orthotopic heart transplantation (OHT). The aim of this study was to identify predictors of the best mid-term outcome in OHT after FF. METHODS Twenty-year multi-institutional retrospective analysis on OHT for FF. RESULTS Between 1991 and 2011, 61 patients, mean age 15.0 ± 9.7 years, underwent OHT for failing atriopulmonary connection (17 patients = 27.8%) or total cavopulmonary connection (44 patients = 72.2%). Modality of FF included arrhythmia (14.8%), complex obstructions in the Fontan circuit (16.4%), protein-losing enteropathy (PLE) (22.9%), impaired ventricular function (31.1%) or a combination of the above (14.8%). The mean time interval between Fontan completion and OHT was 10.7 ± 6.6 years. Early FF occurred in 18%, requiring OHT 0.8 ± 0.5 years after Fontan. The hospital mortality rate was 18.3%, mainly secondary to infection (36.4%) and graft failure (27.3%). The mean follow-up was 66.8 ± 54.2 months. The overall Kaplan-Meier survival estimate was 81.9 ± 1.8% at 1 year, 73 ± 2.7% at 5 years and 56.8 ± 4.3% at 10 years. The Kaplan-Meier 5-year survival estimate was 82.3 ± 5.9% in late FF and 32.7 ± 15.0% in early FF (P = 0.0007). Late FF with poor ventricular function exhibited a 91.5 ± 5.8% 5-year OHT survival. PLE was cured in 77.7% of hospital survivors, but the 5-year Kaplan-Meier survival estimate in PLE was 46.3 ± 14.4 vs 84.3 ± 5.5% in non-PLE (P = 0.0147). Cox proportional hazards identified early FF (P = 0.0005), complex Fontan pathway obstruction (P = 0.0043) and PLE (P = 0.0033) as independent predictors of 5-year mortality. CONCLUSIONS OHT is an excellent surgical option for late FF with impaired ventricular function. Protein dispersion improves with OHT, but PLE negatively affects the mid-term OHT outcome, mainly for early infective complications.