Study on ultra-structural effects caused by Onion yellow dwarf virus infection in ‘Rossa di Tropea’ onion bulb by means of magnetic resonance imaging

\u2018Rossa di Tropea\u2019 onion is a particular pink/red coloured onion cultivated in Calabria region (Southern Italy), representing one of the Italian most important vegetable crops granted with Protected Designation of Origin (PDO) and Protected Geographical Indication (PGI) trademarks. This local cultivar is characterised by a high nutraceutical compounds content showing anti-inflammatory, anti-cholesterol, anticancer and antioxidant properties. As all vegetable crops and Allium spp., \u2018Rossa di Tropea\u2019 onion is affected by several viruses. Among these, the species Onion yellow dwarf virus (OYDV, genus Potyvirus, family Potyviridae), represents the most limiting biotic stress, inducing severe symptoms. OYDV effect on tissues architecture in whole bulbs was investigated using magnetic resonance microimaging (MRI) technique, which allows the interior of samples to be imaged non-invasively and non-destructively and yields quantitative information on physico-chemical parameters describing water mobility (T1 and T2 relaxation times). The use of such tool allowed to determine how OYDV alters plant physiology by inducing water accumulation in bulb tissues as well as causing ultra-structural modifications of cell wall, highlighted by MRI. All these effects resulted in an increase of free water in plant tissues, and consequently relevant water losses during post-harvest storage, seriously affecting bulb quality, marketability and shelf life

The ecology and evolution of diversity and cooperation in bacterial public-goods

Explaining why cooperation exists despite the persistent advantage of cheats has been the focus of much theoretical and empirical attention in biology. Using the bacterium Pseudomonas aeruginosa as a model system for the evolution of cooperation, I investigate two distinct phenomena which may develop our understanding of how cooperation is maintained; 1) tag-based cooperation and diversity; and 2) environmental heterogeneity. The first investigates how diversity in cooperative systems may be a response to the selective pressure exerted by cheating, and how cheats may then regulate communities to maintain diversity: I demonstrate that in competition, tag-based cooperation is able to evade parasitism, provided the public-good is only accessible to producer strains, i.e., the cheat possesses the “wrong” tag. I also demonstrate that cheats can have a marked influence on diversity: In a community of two producer strains with different tags, if a third cheater strain is introduced, it will drive both its own producer and itself extinct. I do not find that the presence of cheats maintains diversity in either structured or unstructured environments, and discuss the possible causes of this. In the second topic of this thesis, I investigate the effect of environmental heterogeneity in resource availability, through space and time, on the evolution of cooperation. Environmental heterogeneity is a ubiquitous feature of natural landscapes, yet its effect on the evolution of cooperation has not been extensively studied. I demonstrate that resource availability heterogeneity, in both time and space, acts to maintain cooperation at higher levels than homogeneous environments of the same total resource value. This effect is due to the covariance between productivity and the cost of cooperation: high resource availability periods and spaces are highly productive, and also incur a relatively lower cost of cooperation.BBSR

Razvoj i implementacija naprednog metoda za unapređenje bezbednosti i optimizaciju radnog okruženja pri manuelnim operacijama

REZIME Savremeni industrijski sistemi se mogu opisati pomoću napredne tehnologije i opreme koju koriste, i često se pogrešno smatra da je ljudski uticaj u takvim sistemima u potpunosti eliminisan. Međutim, čovek je i dalje ključna komponenta za funkcionisanje sistema, obzirom da automatizovane linije nisu eliminisale manuelne operacije. Bez obzira na visok nivo tehnološkog napretka, bezbednost na radu se ne može u potpunosti zagarantovati. Shodno tome, bezbednost i zdravlje na radu ostaje ključna oblast za unapređenje u svakom radnom okruženju. Ergonomija je oblast nauke koji se bavi optimizacijom radnih aktivnosti i ima za cilj smanjivanje broja oboljenja u vezi sa radom. Većina tehnika i metoda za identifikaciju zdravstvenih problema u vezi sa radom i optimizacije radnog okruženja se zasniva na praćenju sprovođenja radnih aktivnosti i izveštavanju o incidentima/akcidentima. Dok, metode za izbegavanje ljudskih grešaka se zasnivaju na obukama i odgovarajućem obrazovanju iz oblasti bezbednosti i zdravlja na radu. Praćenje sprovođenja radnih aktivnosti je subjektivna metoda, a izveštavanje o incidentima/akcidentima može biti veoma značajan parametar za razumevanje problema na radnom mestu. Ipak, činjenice i rezultati ovim metodama se ne dobijaju u realnom vremenu što može predstavljati značajno i važno pitanje za bezbednost na radnom mestu. Sa druge strane, metode za obuku iz bezbednosti i zdravlja na radu, iako je skoro sve vreme neophodna, ne sprečava u potpunosti nastanak incidenata/akcidenata i oboljenja na radnom mestu, što ukazuje na veliki prostor za unapređenje u ovoj oblasti. Ova doktorska disertacija istražuje mogućnosti primene elektrofizioloških merenja kako bi se zabeležilo kognitivno stanje radnika paralelno primenjivajući postojeće standardizovane metode u oblasti ergonomije. Fokus ove doktorske disertacije jeste na upotrebljivosti elektordermalne aktivnosti tokom sprovođenja tri tipa istraživanja i to: dva su vezana za sprovođenje ponavljajućih radnih aktivnosti i treći se odnosi na obuku iz oblasti bezbednosti i zdravlja na radu.ABSTRACT Modern industrial environment can be described by advanced technology and machinery and many believe that human operational tasks are almost eliminated. However, people are still the key component of the processes whilst automation has not eliminated manual operations. Safety in the working place cannot be guaranteed, despite technological advances; accordingly, Occupational Health and Safety remains a crucial sector for improvement in every working environment. Ergonomics deal with the optimization of work design and aims to minimize occupational issues. Most of the techniques for the identification of occupational issues and optimization of working environment are based on task observation and recording incidents. While methods for avoidance of human error are based on training and appropriate safety education. Task observation is a subjective method, and recording of incidents can be an important kind of metrics for understanding the workplace defects. Nevertheless, these methods do not capture the facts in real time and this can be an important issue for the workplace safety. On the other hand, methods of safety training, although are most of the times necessary, do not really prevent from accidents or occupational health problems and still remain an open sector of improvement. This thesis investigates the possibility of electrophysiological recording for the purpose to capture operators’ cognition in parallel with application of ergonomic standardized methods. The study focuses on the usefulness of Electrodermal activity during three case−studies; two of them represent common working tasks of low risk, and one is based on safety training methods. Electrodermal activity has demonstrated to relate discomfort and cognitive status to specific biosignals, therefore, it can be a valid tool for deeply understanding the status of the operators, while they perform cognitive or physical tasks. The core of Electrodermal activity usefulness is that it is a mechanism of sympathetic nervous system and can be used as an index of autonomic reaction to emotions

Genetic manipulation of pyoverdine non-ribosomal peptide synthetases to identify genetic constraints to effective domain recombination

Non-ribosomal peptide synthetases (NRPSs) synthesise small highly diverse peptides with a wide range of activities, such as antibiotics, anticancer drugs, and immunosuppressants. NRPS synthesis often resembles an assembly line, in which each module acts in a linear order to add one monomer to the growing peptide chain. In the basic mechanism of synthesis, an adenylation (A) domain within each module activates a specific monomer. Once activated, the monomer is attached to an immediately downstream thiolation (T) domain via a prosthetic phosphopantheine group, which acts as a flexible arm to pass the substrate between catalytic domains. A condensation (C) domain, upstream to the A-T domains, catalyses peptide bond formation between an acceptor substrate attached to the T domain and a donor substrate attached to the T domain of the upstream module. The peptide remains attached to the T domain of the acceptor substrate, and then acts as the donor substrate for the next C domain. When peptide synthesis reaches the final module, the peptide is released by a thioesterase (TE) domain. The linear mode of synthesis and discrete functional domains within each module gives the potential to generate new products by substituting domains or entire modules with ones that activate alternative substrates. Attempts to create new products using domain and module substitution often result in a loss of activity. The work in this thesis focuses on identifying barriers to effective domain substitution. The NRPS enzyme pvdD, which adds the final residue to the eleven residue non-ribosomal peptide pyoverdine, was developed as a model for domain substitution. The primary benefit for using this model is that pyoverdine creates easily detectible fluorescent products. The first set of experiments focused on testing the limitations of A domain and C-A domain substitutions to alter pyoverdine. Nine A domain and nine C-A domain substitution pvdD variants were constructed and used to complement a P. aeruginosa PAO1 pvdD deletion strain. The A domain substitutions that specified the wild type substrate were highly functional, whereas A domains that specified other substrates resulted in low levels of wild type pyoverdine production. This suggests the acceptor site substrate specificity of the C domain limited the success of A domain substitutions, rather than disruption of the C/A domain junction. In contrast, although C-A domain substitutions in pvdD in some cases synthesised novel pyoverdines, the majority lost function for unknown reasons. The high success rate A domain substitutions (when not limited by the acceptor site specificity of the C domain) suggested the addition of new C domains was a likely cause for loss of function. The second set of experiments investigated whether disrupting the protein interface between C domains and their upstream T domains may cause a loss in function of C-A domain substitutions. However, domain substitutions of T domains were found to have a high rate of success. Therefore, the results thus far confirmed that disrupting interactions of the C domain with A domains or T domains does not have a large affect on enzyme activity. An alternative explanation for the loss in function with C-A domain substitutions is that C domains translocated to a new enzyme are unable to process the new incoming donor peptide chain because of substrate specificity or steric constraints. To develop methods to circumvent limitations caused by the C domain, the final part of this thesis examined acceptor substrate specificity of C domains. Acceptor site substrate specificity was chosen over donor site specificity as it acts on only an amino acid rather than peptide chain. The substrate specificity was narrowed down to a small subsection of the C domain. This was an initial study of C domain substrate specificity, which may guide future development of relaxed specificity C domains

Capacity building and public awareness raising on Nitrates Directive in the countries aspiring to EU accession

The governments of the EU Member States have agreed to potentially enlarge the Union with seven South East European countries (Croatia, the Former Yugoslav Republic of Macedonia, Albania, Bosnia and Herzegovina, Montenegro, Serbia, Kosovo under UN Security Council Resolution 1244) and Turkey. However, these countries will be granted full member status only when all political, legislative and administrative requirements for membership are fulfilled. Transposition, implementation and enforcement of the EU Nitrate Directive (91/676/EC) is one of these requirements. Many policy makers, farmers and consumers from the EU accession countries perceive the Nitrate Directive as a very demanding piece of legislation with little relevance for their countries. Moreover, there is a widespread belief that the Nitrate Directive can potentially limit the competitiveness of their agricultural sector. Limited or partial information and misconceptions about the Nitrate Directive in these countries provokes fear (and sometimes anger), notably by farmers. Consequently, the adoption of the Nitrate Directive receives low political priority. In most countries aspiring to EU membership, there are no other driving forces besides EU accession pushing Governments to adopt the Nitrate Directive. Pressure exerted by health, consumer or environmental protection NGOs hardly exists. Training and education on the Nitrate Directive is poorly covered and addressed by the curricula at higher education organisations, as well as by NGO training programmes. Consequently experts from these countries do not have much opportunity to get acquainted with the Nitrate Directive. The problem persists when these people become governmental officials, extension officers, farm managers, etc., and are supposed to make policy decisions and administer the Nitrate Directive – or advise farmers and manage farms according to EU Nitrates Directive requirements. In order to remedy this problem, several international projects, financed by the Global Environmental Facility fund (administered by the World Bank or the UN Development Programme), the European Commission, and the EU Member State Governments (notably the Netherlands) have been initiated recently. The experience from these projects shows that transfer of information - capacity building and public awareness raising programmes – play a vital role in understanding the rationale behind the Nitrate Directive and in accepting the farming practices it requires. Participatory training, demonstration of nutrient management planning software, on-farm water quality testing with mobile kits, experiments using piezometers/lysimeters and field trials involving various cover crops, buffer strips, etc. accompanied with Web pages, demonstration videos, posters, leaflets, etc. have been shown to be powerful tools to demonstrate the link between water quality and (adverse) agricultural practices. The valuation (“monetisation”) of ecosystem services and environmental costs generated by the fertiliser industry and farming is a newly emerging concept that seems to be a particularly promising tool for awareness raising on the Nitrate Directive. Emerging assessments from the accession countries suggest that hidden costs (public investments and environmental costs) associated with elevated content of nitrates in water can be substantial. Making policy makers and the public at large aware of these costs and of potential savings on them by practising water friendly farming methods (e.g. organic or pastoral farming) can foster the adoption of the EU Nitrate Directive in EU candidate counties and beyond

L’umorismo in pubblicità: Una valutazione dell’impatto dell’umorismo nella pubblicità a scopo sociale.

This thesis aims to discover the motivations leading a company to communicate through humour. Humour is seen as a language in order to understand where, how and when it is usefully used. After an historical and philosophical introduction (Chapter 2), whose purpose is to show the most important schools of thought which developed over the centuries (and sometimes also overlapped) to explain the elusive nature of humour. This study will then explore the modern trends in advertising strategies (Chapter 3) and focus on humorous advertising and the justification of its social and commercial drivers Chapter 4). In Chapter 5, we will study the sense of humour itself and the psycho-social factors of the ideal target. In Chapter 6, we will introduce social communication, taking into account its objectives and themes. Based on past research - we will also study the various codes of expression, analysing the motivations and effects of the humorous language and the “fear arousing” language (Chapter 7). We will then focus our attention in Chapter 8 on the use of humour in social advertising campaigns against smoking through a survey in which we analysed the reactions of 300 university students to a humorous message compared to another one generating anxiety (stressing the negative effects of smoking). The results helped us to draw indications on the efficiency of both means of communication from cognitive, emotional and behavioural points of view. The present analysis has required the use of interdisciplinary analytical tools and in-depth studies in various disciplines: economy, marketing, semiology and sociology, psychology and philosophy. Unfortunately, often these tools could only give an incomplete answer or a reference to other studies. It was therefore inevitable to keep our attention on some aspects and to ignore others: this kind of freedom is certainly possible while treating a theme such as humour.L’obiettivo della tesi è di individuare le motivazioni che spingono un’azienda a comunicare mediante l’umorismo. L’umorismo viene considerato in chiave strumentale al fine di poter trarre indicazioni su dove, come e quando sia più utile optare per tale linguaggio. Dopo un’introduzione storico-filosofica (cap.2) volta a mostrare le principali correnti di pensiero che nei secoli si sono succedute (talvolta sovrapponendosi) per comprendere la sfuggente natura dell’umorismo, lo studio prosegue discutendo le odierne linee di tendenza delle strategie pubblicitarie (cap.3), per poi focalizzarsi sulla pubblicità umoristica e le motivazioni aziendali e sociali che la giustificano (cap.4). In seguito (cap.5) trattiamo il senso dello humour e i fattori psico-sociali del target ideale. Con il capitolo 6 introduciamo la comunicazione sociale, considerandone obiettivi e temi. Quindi affrontiamo i suoi codici espressivi per analizzare, sulla base di ricerche passate, le motivazioni e gli effetti del linguaggio umoristico e di uno di tipo “fear arousing” (cap.7). La nostra attenzione si sofferma poi (cap. 8) sull’utilizzo dell’umorismo nelle pubblicità sociali contro il fumo mediante un’indagine empirica con la quale abbiamo confrontato le reazioni di 300 studenti universitari a un messaggio umoristico con uno che genera ansia (preconizzando le conseguenze del fumo) per trarre indicazioni sull’efficacia delle due comunicazioni dal punto di vista cognitivo, affettivo e comportamentale. La disamina ha sollecitato strumenti analitici interdisciplinari e approfondimenti di diversa natura: economica e di marketing, semiologica e sociologica, psicologica e filosofica, ma spesso questi hanno potuto solo trovarvi un accenno o un rinvio. Ci è stato quindi inevitabile soffermarci su alcuni aspetti e trascurarne altri: una libertà che un tema come l’umorismo di sicuro ammette