Tesi sul tema "Genetically modified"

After ischemic damage in kidney, alternatively activated or M2 macrophages are key players in resolving inflammation and participating in kidney repair. Obtaining this particular phenotype is a process controlled by deregulation of a series of genes. Modulation of those genes could result in controlling macrophages phenotype and functions, aiming new approaches in resolving ARF. The first aim of this study was to identify genes that are related to M2 macrophages that are involved in tissue repair. By functional genomics we have identified the expression of 14 genes crucial for the induction of the endogenous reparative capacity of the kidney that are modulated by the presence of macrophages. We also demonstrated that modulation of Ivns1abp gene can modulate renal repair. More concrete we show that over-expression of the Ivns1abp gene can enhance macrophages regenerative capacity and switch their phenotype towards M2, provoking modulation of their inflammatory state and increasing its resistance against inflammation inputs. We also demonstrated that silencing of the Ivns1abp genet to macrophages leads them to acquire a M1 phenotype and decrease their reparative and phagocytosis capacity. As a last step, we define pathways that regulate the transcription of the Ivns1abp gene. We show that Ivns1abp gene expression in macrophages is regulated by the c-myc transcription factor and modulated by the presence of cytokines determining cell fate.
Después de la lesión isquémica en el riñón, los macrófagos activados alternativamente o M2 no solo participan en la reparación del daño sino son factores clave en la resolución de la inflamación. La obtención de este fenotipo particular es un proceso controlado por la desregulación de una serie de genes. Modulando estos genes, se podría controlar el fenotipo y las funciones de los macrófagos, con el objetivo de nuevos enfoques en el tratamiento de FRA. El primer objetivo de este estudio fue identificar los genes que están implicados en la reparación de tejidos y que están relacionados con los macrófagos M2. Por genómica funcional hemos identificado la expresión de 14 genes cruciales para la inducción de la capacidad regeneradora endógena del riñón y modulados por la presencia de macrófagos. A continuación hemos demostrado que la modulación del gen Ivns1abp puede modular la reparación renal. En concreto hemos demostrado que la sobre-expresión del gen Ivns1abp en macrófagos puede mejorar su capacidad regenerativa y cambiar su fenotipo hacia M2, provocando la modulación de su estado inflamatorio y aumento de su resistencia frente la inflamación. Se demostró además que el silenciamiento del gen Ivns1abp en macrófagos disminuye su capacidad reparadora y fagocítica transformando su fenotipo a M1. Como ultimo, hemos definido las vías que regulan la transcripción del gen Ivns1abp. Hemos demostrado que la expresión del gen Ivns1abp en los macrófagos está regulada por el factor del trascripción c-myc y modulada por la presencia de citoquinas, determinando el destino de la célula.

This study was carried out to analyze tomato samples and tomato seeds, purchased from different food markets of Turkey randomly, for the presence of genetic modification by using PCR method as it allows more specific detection. The DNAs of collected samples were isolated according to CTAB DNA extraction protocol and also with extraction kits. Screening tests of tomatoes were done by targeting 35S promoter, NOS terminator and NptII kanamycin resistance gene with eight different primer sets. Real time PCR is used to confirm 35S and NOS positives results obtained from conventional PCR. In this study, it was observed that 14 out of 35 seed samples, and 14 out of 40 fresh tomato samples which were screened had at least one transgenic element of 35S promoter, NOS terminator and NPTII kanamycin resistance gene indicating the possible presence of genetic modifications. After screening, gene specific studies were carried out for PG, sam-k indicating F type ripening delayed tomato and the 35 1 N lines respectively and cry1Ac genes inserted in 5345-1 insect resistant tomato line. PG and sam-k specific primers were not amplified in any of the samples investigated whereas 18 out of 75 samples were cry1Ac positive and 1 out of 75 samples was sam-k positive. Positives were confirmed by sequence analysis. Additionally, construct specific primers specific to 5345-1 and 35 1 N lines were designed. PCR amplicons indicate the existence of the construct sequence. In order to verify the results, PCR products were sent to sequence analysis

In this thesis I show that macrophages (both cell lines and primary cultures) can be transfected by recombinant adenoviruses expressing -galactosidase (A-gal/Av1Bng), that the macrophages become activated by the transfection process and can be easily manipulated to localise to inflamed glomeruli after direct injection into the renal artery of rats with an experimentally induced glomerular inflammation caused by nephrotoxic nephritis. The injection of transfected macrophages reduces the severity of injury in this model of glomerulonephritis as shown by a reduction in the degree of albuminuria. This approach provides a favourable system for gene delivery in inflammatory disease and shows that both the functional properties of the transfected macrophage as well the transgene it is engineered to produce are relevant for in vivo gene transfer. This approach has also been used to determine the effect of macrophages expressing active TGF-1 on the development of glomerular inflammation. TGF-1 expressing macrophages localised efficiently to inflamed glomeruli and produced the cytokine in vivo. These cells produced a reduction in the level of albuminuria compared to unmodified disease but not in comparison to injection of macrophages transfected with adenovirus expressing -galactosidase. In addition there was no alteration in the infiltration by ED1 positive macrophages. Thus TGF-1 expressed in this manner appears unable to significantly modulate glomerular inflammatory disease and the potential reasons for this are discussed. The system I have developed of macrophage transfection and delivery provides a valuable approach to study and modulate inflammatory disease.

With the adoption of biotechnology in many agricultural products with first-generation biotechnology traits such as increased pest resistance, greater herbicide resistance, and increased yields the growers have accepted them. The next wave of biotech crops have second-generation traits, such as improved nutrient content, extended shelf life, reduced pesticide and herbicide application (a consumer demanded trait), and better taste. Will these consumer benefits offset any concern that the consumer has regarding biotechnology? What are those benefits and how should the information be communicated to the consumer? Three focus groups give insight to the proposed questions. The focus groups were done in three California cities, with participants screened to be: 18-65 years of age, the primary shopper for the household, and with an education level up to a bachelor’s degree. We found that the consumer has little knowledge of biotechnology, but that they assume any concern over these products is reduced if the grocery store or point of purchase is a reputable location. The consumer does look for added utility in products, but they are not willing to pay more unless they understand the production of biotechnology developed products. The consumer feels that there should be labeling of these products, but will likely purchase the least expensive option.

The debate around genetically modified food and crops has proved to be complex and far-reaching, involving diverse stakeholder groups and many issues. Although the extent of global uptake of GM crops has been substantial (23 countries and 114.65 million hectares by 2007), it is significant that four countries are responsible for 86% of all GM plantings, and that a number of key food markets (for example the EU and Japan) remain largely "GM-free‟. This suggests that there is reluctance on the part of many countries to embrace GM technology. There are likely many reasons for this, but one significant issue is that of the perception of the risks associated with the technology. There is a distinction between risk that exists in the world and that can be measured (objective risk) and risk that is perceived by an individual to exist and that is constructed by them based on their values and preferences. When technical measurement of actual risks is not possible, peoples‟ own perceptions of risks become important. This thesis aims to investigate the topic of risk perceptions associated with GM food and crops. Different stakeholder groups have been targeted, and a range of methodologies from a variety of disciplines have been employed to investigate what factors can be shown to influence risk perception. A range of factors were identified from existing literature, as having potential impact on risk perceptions. A number of these were investigated, some of which were found to have some influence on levels of risk perception. Results demonstrate that factors influencing peoples‟ perceptions of risk relating to GM food and crops, include the uncertainty associated with the technology, and trust in regulators, policy makers and others with control over the future development of the technology. Other factors found to be important to levels of risk perception held by different stakeholder groups, were a range of socio-demographic and cultural variables, the relationship between perceived risks and benefits, the equity of impacts, and the influence of third parties. There are a number of implications for the development of the GM debate arising from the findings. First, as there are socio-demographic and cultural factors linked to the perceptions of risk associated with GM technologies in food and agriculture, it is important to recognise that different people will react differently to the technology. Specifically, results from this thesis show that it may be that men, those who are more highly educated, those with a less ecocentric worldview, and those living in urban areas, are likely to respond more favourably to targeted promotional campaigns. As regards the farming community, results show that the first farming adopters are likely to be those who are both owners and tenants, not in an urban fringe location, potato growers, and not barley growers. Second, this thesis provides evidence that third parties are particularly important to farmers, thus it is crucial to recognise that there is potentially a long chain of action and reaction amongst many different stakeholders and actors impacting on farmers' levels of risk perception, and hence willingness to adopt the technology. Third, results from this research demonstrate that the linked issues of the relationship between risks and benefits, and the equity of (positive and negative) impacts, require that all stakeholders are content that they will receive a share of the benefits (if any) to be derived from the technology, and that neither they nor any other group of stakeholders are unduly impacted by the risks or negative impacts (if any) of the technology. Important here is the recognition that perceptions are as important as actual impacts. Fourth, the issue of trust has been shown by the results obtained by this research to be extremely important to peoples' perceptions of risk. It can be concluded that trust is of wider social and political importance that relates to the need to ensure greater democratisation of decision-making in order to re-establish trust in authorities. In the case of GM food this may require a rethinking of the EU legislation relating to the technology. This also relates to point below about the delivery of messages and education. Information sources must be trusted by those at whom the messages are aimed. More importantly though, if people are to trust decision making processes, there needs to be stakeholder involvement at an early stage of decision making, that allows some impact on decisions taken. In the case of the GM debate it may indeed be too late as decisions about the technology, its applications, the regulatory processes and its inclusion within the food chain are well established. Perhaps the best that can be hoped for is that lessons will be learnt and applied to future technology developments of relevance to the food chain, such as, nano-technology. Finally, this thesis has shown that uncertainty is central to peoples' perceptions of risk. This could be addressed through a combination of additional research into what is uncertain to people, the impacts and implications of the technology, more effective dissemination of existing knowledge, and impartially delivered messages and education strategies from trusted sources that address the concerns that people have about the technology. Importantly however there must be an acknowledgement that uncertainty is not restricted to "knowledge deficit‟ but encompasses the scientific uncertainties inherent within the technology, and is framed by the social and cultural values of those whose views are considered. This thesis uniquely targeted diverse groups and employed a combination of different methods from a variety of disciplines. By doing this the study has increased understanding of the views of two groups (campaigners and farmers) who are crucial to the uptake of the technology, and who are seldom researched in the area of attitudes to GM technologies. The diversity of groups, methods and disciplines brought together in this thesis is important because the issue of GM has proved to be complex and far-reaching, and previous discussions of risk perceptions have been complex and disjointed. All groups investigated here are stakeholders in the process, and as such their views and concerns relating to risk perceptions of GM technologies ought to be taken into consideration.

Public controversy over genetically modified (GM) foods illustrates the increasing complexity of the governance of technological innovation. In the light of public displays of ambivalence towards biotechnology, corporations are paying greater attention to societal concerns over the innovation of new technologies. This thesis asks how those corporations involved in the development and commercialisation of GM foods have understood and responded to recent public controversy over biotechnology in Europe and North America. Using approaches drawn from geography and science and technology studies, this thesis is concerned with concepts of citizenship and also with corporate identities in the context of technological controversy. This thesis focuses particularly on the ways that corporations have understood the emergence of citizen-like demands to participate in the governance of corporate innovation. The research for this thesis adopts an ethnographic approach to studying corporate engagements in public controversy. It uses the analysis of corporate texts, interviews and participation in meetings at which corporate presentations were made about GM foods. The empirical material for this thesis refers to three corporations: DuPont, Monsanto, Unilever and two agricultural biotechnology public relations groups: the Council for Biotechnology Information in the USA, and the Agricultural Biotechnology Council in the UK. This thesis argues that corporations have innovated new institutional forms of engagement with the public over biotechnology. These engagements have configured the 'socially responsible corporation* and the 'consumer-citizen' as actors in more democratic forms of corporate innovation. This thesis concludes that these new forms of governance are partial, fragile and contested, but that they offer potential avenues for further public debate about the governance of corporate innovation.

This thesis develops biophysically-based data-driven mathematical models of intracellular calciumdynamics in ventricularmyocytes for both normal and genetically modified mouse hearts, based on species- and temperature-consistent experimental data. The models were subsequently applied to quantitatively examine the changes in calcium dynamics in mice with cardiomyocyte-specific knockout (KO) of the cardiac sarco/endoplasmic reticulum ATPase (SERCA2) gene, to determine the contributing mechanisms which underlie the ultimate development of heart failure in these animals. In Chapter 1, with emphasis on calcium dynamics and calcium regulation in heart failure, an overview of cardiac electrophysiology, excitation-contraction coupling and mathematical models of cardiac electrophysiology is provided. In Chapter 2, models of calcium dynamics in the ventricular myocytes from the C57BL/6 mouse heart at a physiological temperature is developed and validated based on species- and temperature-consistent measurements. In Chapter 3, the C57BL/6 model framework is re-parameterised to experimental data from the control and SERCA2 KO mice at 4 weeks after gene deletion. The models are then used to quantitatively characterise changes in calcium dynamics in the KO animals and the role of the compensatory mechanisms. In Chapter 4, the model framework is extended to include differential distributions of ion channels in the sarcolemma and the calcium dynamics in the sub-sarcolemmal space, with parameters in these sub-components fitted to experimentally measured calcium dynamics from the control and KO cardiomyocytes at 7-week after gene deletion. Finally in Chapter 5, conclusions are drawn, the limitations of this study are discussed, and the future extensions to this study are described.

In recent years, foods produced by genetic engineering technology have been on the world food market. The biosafety aspects, regulations, and labelling of these foods are still contentious issues in most countries. It is necessary to have approval for the use of GMOs in the production of food. Thus, detection and quantification of GMOs play crucial role for developing regulations on GM foods. In this study, raw and processed maize samples were analysed for genetic modification using a DNA based detection method, the Polymerase Chain Reaction. Ten raw food and 18 processed maize food including maize flour, starch, corn flakes, maize chips were collected from different markets located in different places in Turkey. The samples were examined for the presence of genetic elements located in the majority of transgenic crops such as NOS terminator, CaMV 35S promoter, kanamycin resistance (KanR) gene, using conventional PCR with oligonucleotide sets targeting to novel genes. Furthermore screening was conducted via Real-Time PCR assay for NOS terminator and 35S promoter. For confirming the presence of Bt11 maize lines event specific primers were utilised. Quantification of Bt11 maize lines were performed via Real-Time PCR. The result indicates that foreign genetic elements were found in all analysed raw material. In six out of 10 raw material, presence of Bt11 gene were identified. GMO detection was also possible for maize flour and starch, however in processed material as corn starch, corn flakes, corn chips and pop corn, transgenes were not detected.

The genetically modified foods controversy is a dispute over the relative advantages and disadvantages of food derived from genetically modified organisms, genetically modified crops used to produce food and other goods, and other uses of genetically modified organisms in food production. The dispute involves consumers, biotechnology companies, governmental regulators, non-governmental organizations and scientists. The key areas of controversy related to genetically modified (GM) food are: risk of harm from GM food, whether GM food should be labeled, the role of government regulators, the effect of GM crops on the environment, the impact of GM crops for farmers, including farmers in developing countries, the role of GM crops in feeding the growing world population, and GM crops as part of the industrial agriculture system. When you are citing the document, use the following link http://essuir.sumdu.edu.ua/handle/123456789/33677

As economies develop, novel products are created and markets for these products arise. Genetically modified food (GMF) is an example of such a novel product and provides economists with the opportunity to investigate an infant market. Of particular interest with GMF is the impact of consumer reactions on the market. The response of consumers to GMF and their willingness to pay for it has emerged as an important factor in the development of this technology. This research investigates these consumer responses. Prior research suggests that two aspects of consumer behaviour may be relevant for the GMF market. First, consumers may react differently to different types of GMF, so that some products are potentially more economically viable. Secondly, some consumers appear to prefer not having GMF at all. Consumer behaviour is often framed according to neoclassical economic theory. Consumer preferences over goods and the attributes of those goods are generally held to have certain properties. The aspects of consumers’ reactions to GMF noted above, however, may be in conflict with two properties of preferences in neoclassical theory. First, preferences over food attributes are not separable, but may interact with each other. Secondly, some consumers may have preferences regarding GMF that are not continuous. As a result, aggregate impacts of introducing GMF may be difficult to measure, which raises a third issue for investigation, aggregation. Finally, an alternative model of consumer behaviour is bounded rationality, which theorises that choices may be discontinuous as a result of specific protocols. It also suggests that consumers seek to make good-enough choices, rather than attempting to maximise their satisfaction. Thus, optimisation or maximisation is the fourth issue considered in this thesis. In order to investigate these properties of consumers’ preferences, a choice experiment survey was developed. The strength of a choice experiment for examining these issues is its focus on the impact of each product attribute on a respondent’s choices. Thus, it may be possible to identify potentially discontinuous choice patterns and to identify choices affected by interactions between GM technology and other food attributes. Results from a neoclassical analysis of the survey data suggest that some consumers consider the type of benefit created with GM technology in making their choices. In addition, one-quarter to one-half of respondents may have had discontinuous preferences with respect to GMF. Reactions to GMF appear related to respondents’ attitudes, but not to socio-economic or demographic descriptors. As a result, aggregate measures of the impact of GMF may not fully account for consumers’ responses. A boundedly rational model also has reasonable goodness of fit, and may provide a different perspective on consumer behaviour. It is hoped that the results of this research provide a better understanding of consumer behaviour regarding GMF and, by extension, of the process of consumer adoption of novel products. It is further hoped that this attempt to incorporate choice protocols into discrete choice analysis will provide a useful example for further research.

As economies develop, novel products are created and markets for these products arise. Genetically modified food (GMF) is an example of such a novel product and provides economists with the opportunity to investigate an infant market. Of particular interest with GMF is the impact of consumer reactions on the market. The response of consumers to GMF and their willingness to pay for it has emerged as an important factor in the development of this technology. This research investigates these consumer responses. Prior research suggests that two aspects of consumer behaviour may be relevant for the GMF market. First, consumers may react differently to different types of GMF, so that some products are potentially more economically viable. Secondly, some consumers appear to prefer not having GMF at all. Consumer behaviour is often framed according to neoclassical economic theory. Consumer preferences over goods and the attributes of those goods are generally held to have certain properties. The aspects of consumers' reactions to GMF noted above, however, may be in conflict with two properties of preferences in neoclassical theory. First, preferences over food attributes are not separable, but may interact with each other. Secondly, some consumers may have preferences regarding GMF that are not continuous. As a result, aggregate impacts of introducing GMF may be difficult to measure, which raises a third issue for investigation, aggregation. Finally, an alternative model of consumer behaviour is bounded rationality, which theorises that choices may be discontinuous as a result of specific protocols. It also suggests that consumers seek to make good-enough choices, rather than attempting to maximise their satisfaction. Thus, optimisation or maximisation is the fourth issue considered in this thesis. In order to investigate these properties of consumers' preferences, a choice experiment survey was developed. The strength of a choice experiment for examining these issues is its focus on the impact of each product attribute on a respondent's choices. Thus, it may be possible to identify potentially discontinuous choice patterns and to identify choices affected by interactions between GM technology and other food attributes. Results from a neoclassical analysis of the survey data suggest that some consumers consider the type of benefit created with GM technology in making their choices. In addition, one-quarter to one-half of respondents may have had discontinuous preferences with respect to GMF. Reactions to GMF appear related to respondents' attitudes, but not to socio-economic or demographic descriptors. As a result, aggregate measures of the impact of GMF may not fully account for consumers' responses. A boundedly rational model also has reasonable goodness of fit, and may provide a different perspective on consumer behaviour. It is hoped that the results of this research provide a better understanding of consumer behaviour regarding GMF and, by extension, of the process of consumer adoption of novel products. It is further hoped that this attempt to incorporate choice protocols into discrete choice analysis will provide a useful example for further research.

Neisseria meningitidis colonises the human nasopharynx and can invade through the epithelium to cause rapidly progressing, potentially fatal disease. Carriage of the commensal Neisseria lactamica correlates with decreased colonisation with N. meningitidis and reduced invasive meningococcal disease (IMD), making N. lactamica a candidate for an anti-meningococcal vaccine or probiotic. Intranasal inoculation of N. lactamica showed a good safety profile, but only 34% of those challenged with this species became colonised. Intramuscular injection of N. lactamicaderived outer membrane vesicles (OMVs) demonstrated their potential as a mucosal adjuvant but with limited anti-meningococcal immunogenicity. This project investigated the use of genetically modified (GM) N. lactamica to enhance the potential of this organism to protect against IMD. N. lactamica previously proved resistant to genetic modification. Here we showed that a significant barrier to transformation is the activity of restriction enzymes, which was partially overcome by the incorporation of exclusively hypermethylated cytosine residues into donor DNA, by bespoke PCR amplification. Although it was observed that N. lactamica Y92-1009 is highly resistant to heterologous genetic recombination in vivo, in vitro and in biofilms, this technique allowed us to accurately target genetic modifications to various chromosomal loci in this strain. Multiple factors affecting the efficiency of transformation were identified, including the extent of homology between donor and chromosomal DNA and the number of DNA uptake sequences. Transformation was found to be dependent on the presence of type 4 pili, which were also required for efficient interaction of N. lactamica with epithelial cells. Meningococcal immunogen PorA was selected for the proof of principle that heterologous antigens could be expressed in N. lactamica. With the view to progress this strain to human challenge, the protein was introduced in an antibiotic-resistance-free manner, using the deletion and re-introduction of β-galactosidase as a marker of transformation on X-Gal-containing agar. PorA P1.7,16 was expressed at modest levels but the expression of other PorA types was unsuccessful. PorA P1.7,16 was presented on the surface of bacteria and on OMVs. Intraperitoneal injection of mice with PorA-containing OMVs induced production of serum bactericidal antibodies (SBA) that were active against both N. meningitidis H44/76 and MC58. A genetic enhancer sequence was identified upstream of PorA in N. meningitidis and was used to modulate PorA expression in N. lactamica. In order for N. lactamica to tolerate phase variable expression of PorA, the enhancer sequence was truncated and expression reduced. At this reduced level, a panel of three PorA-expressing strains of N. lactamica were assessed for progression to human challenge. Whilst strains were comparable, genetically stable and antibiotic susceptible, the low-level PorA expression meant that phase variation could not be detected and OMVs derived from these strains did not induce SBA activity in mice. This thesis presents the first steps in developing GM strains of N. lactamica with the potential to reduce nasopharyngeal colonisation with N. meningitidis and to vaccinate against invasive meningococcal disease.

Dissertation (PhD(Agric))--University of Stellenbosch, 2005.
ENGLISH ABSTRACT: Considerable efforts have been made to improve strains of the wine yeast Saccharomyces cerevisiae through the use of genetic engineering. Although the process is well defined, globally there is much resistance towards the use of genetically modified organisms (GMOs), primarily because little is known about their environmental fate and their potential effect on naturally occurring organisms. The public concern is mainly focused on the uncertainty associated with the impact of the deliberate or accidental release of a GMO into the environment. As a consequence, thére is an urgent need to assess the potential risks involved with the use of this new technology. For the eventual global acceptance of any GMO, it is imperative that the consumer must be convinced that it is ultimately safe for human consumption and the environment. In order to achieve this, certain risk assessment procedures must be performed on each and every GMO that is planned to be released into the environment. Although some of the genetically modified (GM) yeasts that have been developed comply with the strict legislation of most countries and have been cleared by regulatory authorities for commercial use, GM yeasts have not, as yet, been used for the commercial production of GM bread, beer or wine. Nevertheless, the use of GM yeasts in the market appears imminent and there is an urgent need to assess and address the perceived health and environmental risks associated with GM foods. The overall objective of this research was to evaluate key environmental issues concerning the use of GM yeasts. The focus was on comparing the behaviour of specific parental and GM yeast strains in model systems in order to determine whether the GM strains may have any selective advantage, which could lead to their spreading. Specifically, it involved monitoring of the growth behaviour of selected GM yeasts within a vineyard microbial community and in fermentations, as well as the interaction of these yeasts with sand and glass surfaces in an aqueous environment. The GM yeasts under investigation were recombinant strains of a well-known, industrial strain of S. cerevisiae VIN13 expressing an a-amylase (designated GMY1); an endo-p-1,4-glucanase and endo-p-xylanase (designated GMY2); and a pectate lyase and polygalacturonase (designated GMY3). The GM yeasts were mist-inoculated onto individually-contained blocks consisting of one-year old grapevines in a secluded glasshouse environment. Specifically, the numbers and dynamics of GM yeast survival, as well as the effect of an introduced GM yeast on the yeast community dynamics and numbers, were investigated. Overall, it was found that the most prevalent wild yeasts isolated from the grapevines were Rhodo torula, Yarrowia lipolytica, Pichia and Candida spp. VIN13 and the GM yeasts did not affect the overall ecological balance of the microflora on the grapevines. Wild strains of S. cerevisiae were seldom isolated from the grapevines. With a few exceptions, the overall detection of GM yeasts was numerically limited. Co-inoculation of (VIN13+GMY1) and (GMY1+GMY2) revealed detection approximately in the same ratio at which they were inoculated, with small differences in the order of GMY2>GMY1 >GMY3. GM yeasts were rarely isolated from bark and soil samples. Spontaneous fermentation of the grapes harvested from the different treated blocks indicated that the GM yeasts survived on the berries, that the natural fermenting ability of VIN13 was conserved in the recombinant strains, and that the GM yeasts did not have any competitive advantage. The soil environment forms an important part of the biosphere and the transport and attenuation of a GM yeast in this matrix will to a large extent affect their ultimate fate in the environment. In soil, microorganisms either occur as suspended cells in pore water or as biofilms on soil surfaces. Although less extensive than a typical soil yeast, Cryptococcus, epifluorescent staining of biofilms confirmed that VIN13 and GMY1 were capable of existing in a biofilm mode on sand granules and glass. Data on effluent numbers detected in flow cells indicated that GMY1 had no advantage due to the genetic modification and had the same reproductive success as VIN13. These strains either had no difference in biofilm density or GMY1 was less dense than VIN13. When co-inoculated, GMY1 had no negative influence on the mobility of Cryptococcus through a sand column, as well as the ability of Cryptococcus to form biofilms. Furthermore, it was found that GMY1 did not incorporate well into a stable biofilm community on glass, but did not disrupt the biofilm community either. This is the first report of the assessment of the fate of GM strains of VIN13 that are suitable for the wine and baking industry. The investigation of the GM yeasts in this study under different scenarios is a good start to an extensive and necessary risk assessment procedure for the possible use of these GM yeasts in the industry. This study could lead to the provision of much-needed scientific and technical information to both industry and regulating bodies. The outcome of this research is also intended to serve as a basis for information sharing with public interest groups.
AFRIKAANSE OPSOMMING: Aansienlike pogings is reeds aangewend om rasse van die wyngis, Saccharomyces cerevisiae, deur middel van genetiese manipulering te verbeter. Alhoewel hierdie proses goed gedefinieerd is, is daar wêreldwyd heelwat teenkanting teen die gebruik van geneties gemanipuleerde organismes (GMO's). Dit is hoofsaaklik te wyte daaraan dat so min bekend is oor hul lot in die omgewing en hul potensiële effek op die organismes wat natuurlik voorkom. Die publiek is veral besorg oor die onsekerheid verbonde aan die bestemde of toevallige vrylating van 'n GMO in die omgewing. Gevolglik is daar 'n dringende behoefte om die potensiële risiko's in die gebruik van hierdie nuwe tegnologie te bepaal. Dit is van uiterste belang dat die verbruiker oortuig sal word van die veiligheid vir menslike gebruik en die omgewing voordat enige GMO uiteindelik wêreldwyd aanvaarbaar sal word. Om dit te kan bereik sal sekere risiko-bepalende prosedures toegepas moet word op ieder en elke GMO wat beplan word om vry gelaat te word in die omgewing. Alhoewel sommige van die geneties gemanipuleerde (GM) giste aan die streng wetgewing van die meeste lande voldoen en deur die owerhede vir kommersiële gebruik goedgekeur is, word GM-giste nog steeds nie vir die produksie van GM brood, bier of wyn gebruik nie. Ten spyte hiervan, blyk die gebruik van GM-giste onafwendbaar te wees en is daar dus 'n dringende behoefte om die voorspelde gesondheids- en omgewingsrisiko's wat met GM voedsel geassosieer word, aan te spreek. Die oorhoofse doel van hierdie navorsing was om belangrike omgewingskwessies aangaande die gebruik van GM-giste te evalueer. Die fokus was op die vergelyking van die gedrag van spesifieke oorspronklike gisrasse en GM-gisrasse in modelsisteme sodat daar bepaal kon word of die GM-gisrasse enige selektiewe voordele het wat moontlik tot hulonbeheerde verspreiding in die natuur sou kon lei. Dit het spesifiek die monitering van die groei van geselekteerde GMgiste binne 'n mikrobiese gemeenskap op wingerd en in fermentasies behels, asook die interaksie van hierdie giste met grond en glas oppervlaktes in 'n wateromgewing. Die GM-giste wat in hierdie studie gebruik is, was rekombinante rasse van 'n bekende industriële ras van S. cerevisiae, VIN13, wat geneties gemodifiseerd was om 'n a-amylase (aangedui as GMG1); 'n endo-p-1,4-glukanase en 'n endo-B-xilanase (aangedui as GMG2); en 'n pektaatliase en 'n poligalaktorinase (aangedui as GMG3) uit te druk. Die GM-giste is op afsonderlike blokke van eenjaaroue wingerdplante binne-in 'n beskutte kweekhuis gesproei-inokuleer. Daar was spesifiek na die selgetalle en dinamika van die oorlewende GM-giste gelet, asook wat die invloed was van die inokulasie van 'n GM gis op die selgetalle van die natuurlike gisgemeenskap. Daar is bevind dat die wildegiste Rhodotorula, Yarrowia Iipolytica, Pichia en Candida spp die gereeldste van die wingerd geïsoleer is. VIN13 en die GM-giste het nie die ekologiese balans van die natuurlike mikrobiese populasie op die wingerd versteur nie. Wilde rasse van S. cerevisiae is selde geïsoleer vanaf die wingerd. In die meeste gevalle is daar bevind dat wanneer GM-giste opgespoor is, hulle in lae selgetalle voorgekom het. Waar giste saam geïnokuleer was, was die opsporing van (VIN 13+GMY1) en (GMY1 +GMY2) ongeveer in dieselfde verhouding as waarin hul geïnokuleer was, terwyl klein verskille in die orde van GMY2>GMY1 >GMY3 opgemerk is. GM-giste is selde vanaf bas- en grond-monsters geïsoleer. Spontane fermentasies van druiwe wat geoes vanaf die verskillende behandelde blokke is, het daarop gedui dat die GM-giste wel op die druiwe oorleef, dat die natuurlike vermoë van VIN13 om te kan fermenteer in die gemodifiseerde gisrasse behoue gebly het en dat die GM-giste geensins deur die genetiese modifikasies bevoordeel was nie. Grond is 'n belangrike deel van die biosfeer en die verspreiding en aanhegting van 'n GM-gis in hierdie matriks sal sy algehele lot in die omgewing tot 'n groot mate beïnvloed. In die grond kom mikroorganismes as gesuspendeerde selle in poriewater of as biofilms op die oppervlaktes van grond voor. Alhoewel biofilmvorming van VIN13 en GMG1 swakker was as in die geval van 'n tipiese grondgis, Cryptococcus, het epifluoresserende kleuring van hierdie S. cerevisiaegiste bevestig dat VIN13 en GMG1 in staat was om as biofilms op sandkorrels en glas te oorleef. Gebaseer op seltellings in vloeiseluitlaat, kon daar afgelei word dat GMG1 geen selektiewe voordeel geniet het as gevolg van die genetiese modifikasie nie en dat die gis net so reproduktief was as VIN13. Hierdie gisrasse het geen verskil in biofilmdigtheid getoon nie of die biofilmvorming van GMG1 was minder dig as die van VIN13. Wanneer GMG1 saam met Cryptococcus geïnokuleer was, het GMG1 geen negatiewe invloed op die beweeglikheid van Cryptococcus deur 'n sandkolom gehad nie en die vermoë van Cryptococcus om biofilms te vorm is ook nie beïnvloed nie. Daar is verder ook bevind dat GMG1 nie goed binne-in 'n gestabiliseerde biofilmgemeenskap op glas geïnkorporeer het nie, maar dat die gis ook nie die biofilmgemeenskap versteur het nie. Hierdie studie verteenwoordig die eerste ondersoek ooit oor die lot, oorlewing en groeigedrag van GM-wyngiste in biologies-afgesonderde wingerd-, fermentasie-, modelgrond- en modelwater-ekosisteme. Die bestudering van hierdie GM-giste onder verskillende omgewingstoestande in afgeslote ekosisteme lê 'n stewige basis vir verdere ondersoeke en die ontwikkeling van omvattende en noodsaaklike risikobepalingsprosedures betreffende die moontlike toekomstige gebruik van GM-giste in die industrie. Hierdie studie baan die weg tot die verkryging van noodsaaklike wetenskaplike en tegniese inligting oor die veiligheidsaspekte rakende GM-wyngiste en dit kan van groot waarde vir die industrie, owerhede en verbruikerspubliek wees.

The long-term implications of deregulation of glucose metabolism, manifesting itself as diabetes is significant. Whilst treatments for the disease are progressing, often they require multiple daily insulin injections. This may be difficult for diabetics in poorer nations. This thesis examines the possibility of utilising keratinocytes and melanocytes transfected with either normal or furin-modified proinsulin constructs, to process and secrete mature insulin via secreted protein pathways. The advantage of this approach is that it may allow normalisation of basal glucose levels, which in turn may prevent or delay the onset of many of the complications associated with diabetes. The results confirmed that keratinocytes and melanocytes are able to process and secrete mature insulin from a furin-modified proinsulin construct, thus validating this approach and paving the way for skin grafts of genetically modified cells to be used as a possible treatment for diabetes. Whilst the results regarding the ability of these cells to process a normal proinsulin construct are inconclusive, they highlight areas of interest that could help to resolve this issue
Master of Science (Hons.)

Methods currently available for the detection and enumeration of genetically modified micro-organisms in the environment include culturing methods, direct microscopic detection and nucleic hybridization techniques. The aim of this project was to develop luminescence as a molecular based-marker system in Pseudomonas fluorescens. The lux genes, originally isolated from Vibrio fischeri, were introduced into Ps. fluorescens on plasmid vectors and on the chromosome. The efficiency of these two strategies for the detection of Ps. fluorescens in soil was assessed. Luminometry was used to estimate biomass concentration during growth. The sensitivity of luminescence detection was greater for the plasmid marked Ps. fluorescens in both liquid culture and soil, however, cellular light output was less closely linked to biomass concentration. Enumeration of cells by luminometry was only possible for growing cells as light output is correlated with microbial activity. The lux chromosomal marker was stable in liquid culture for at least 200 generations and in soil for up to 135 days. The plasmid borne lux genes had a half-life of 20 generations in liquid culture. After inoculation in sterile soil, plasmid loss was only observed during cellular growth. The frequency of transfer of the lux genes from Ps. fluorescens, by conjugation and transformation, was assessed in liquid culture. Mobilisation of these genes by three self transmissible plasmids was negligible due to the instability of these vectors in this host. Transformation of Ps. stutzeri with lux genes, by cell contact, was at frequencies below levels of detection. Luminescence provided a valuable marker for tracking pseudomonads in soil. Detection of marked strains by luminometry provided a sensitive, rapid and non-extractive technique for enumeration of growing cells and measurement of microbial activity. As the chromosomally encoded lux genes were stable, regardless of growth conditions, and emitted sufficient levels of light to enable visual enumeration of colonies by eye, this was considered the best system for long term risk assessment studies.

Introduction: The Genetically Modified (GM) food, which is one of the fruit of the modern biotechnology, is closely related to people's lives. GM food, specifically, GM crops, also known as biotech food, are produced from genetically modified organisms (GMO), which use genetic engineering techniques to introduce, recombine and modify DNA. The safety of GM food still do not have final conclusion at present. Although GM food has been introduced into China for over 15 years, many of the surveys show that Chinese consumers‘ knowledge of GM food is relatively low comparing with other countries. In 2002, China‘s Ministry of Agriculture promulgated three regulations to manage the GMOs in China. The attitudes and acceptance of market for GM food have direct impact on the development of genetic engineering technique and government‘s policy making. This review aims to investigate cognition and attitudes of GM food among Chinese consumers; to investigate how the consumers react to the GM food labeling policy; to find out how factors such as knowledge about transgenic information, price of GM food would affect the consumption of GM food. Methods: Relevant studies published between January 2002 and May 2013 were searched and identified through NCBI, CNKI, and Google Scholar with a combination of keywords, such as “GM”, “China”, “attitude”, “knowledge”, and “willingness” both in English and Chinese. Studies regarding the average knowledge level, acceptance and willingness-to-pay (WTP) for GM food among Chinese consumers, and factors affecting the WTP were included. Results: Of 1032 papers identified, 9 articles fulfilling the selection criteria were included in this systematic review. Among the 9 articles, 7 were written in English, 2 in Chinese. Awareness and knowledge of GM food among Chinese consumers were still not satisfying. Given the potential risks, the public tended to hold divergent attitudes to GM food, which had significantly influenced the WTP. Socio-economic factors such as the number of children, and external factors such as information and price also affected the WTP. Among all the factors, positive attitudes (including willingness-to-accept WTA), and positive information about GM food have significant positive influence on the WTP of GM food, while consumer‘s number of children, price of GM food and negative information about GM food would reduce the purchase intention. Discussion: Positive attitudes (including willingness-to-accept WTA), and positive information have significant positive influence on the WTP of GM food, while increasing number of children and price of GM food and negative information would reduce the purchase intention. It is necessary to strengthen the comprehensive and objective propaganda of GM food and transgenic technology; enhance the management of GM food; plan and develop GM food industry with focus on low-income consumers for they are more willing to buy GM food.
published_or_final_version
Public Health
Master
Master of Public Health

The effect of L. terrestris on the transport of marked bacterial inocula in the absence of percolating water was investigated using repacked soil microcosms over a 10 day period. The microcosms consisted of cylindrical cores containing loamy sand. The bacterial inoculum was applied in filter paper disks to the surface of soil cores containing L. terrestris and to control cores. Destructive analysis of cores was carried out 5 and 10 days after inoculation to facilitate enumeration of marked bacterial inocula at 3 depths in the soil. Significantly greater vertical transport of marked bacteria occurred in cores containing L. terrestris. The effect of L. terrestris on the dispersal of marked bacteria in the presence of percolating water was investigated using both repacked soil cores and larger intact cores over a one month period. Bacterial inocula were again applied in filter paper disks to the surface of cores and the cores subjected to simulated rainfall events at 3 day intervals. Concentrations of marked bacteria in leachate were determined and destructive harvests of cores carried out after 24 days. The results from repacked soil cores demonstrated that cores containing earthworms were associated with early breakthrough, higher percentage recovery and greater numbers of marked bacteria in the leachate than control cores without earthworms. Results from intact soil cores demonstrated that cores containing earthworms were associated with different bacterial leaching patterns, higher percentage recovery and greater numbers of bacteria in the leachate than control cores. For both repacked and intact cores, the presence of earthworms in cores led to transport of marked bacteria to greater depths in the soil. Plaster of Paris impregnation of repacked and intact cores highlighted the importance of earthworm burrowing in providing pathways for bypass water flow and hence bacterial transport through soil. Results from the use of interdisciplinary methods developed and adapted in this study identify the importance of earthworms in the fate of microbial inocula in soil and the potential for use of earthworms in biotechnology.

Survival of P. fluorescens 10586s FAC510 was studied in liquid culture and soil microcosms in the presence of the ciliate C. steinii and the flagellate Cercomonas sp. Both protozoa caused a reduction in bacterial viable cell concentration, however the ciliate caused a greater decrease in abundance. Starvation of the bacterial prey caused a reduction in grazing rates of both the ciliate and flagellate, indicating the lower nutrional quality of starved as opposed to non-starved cells. Manipulation of the physiological state of prey cells, by starvation, had much the same effect as observed when bacterial prey resources are reduced. Luminescence provided a valuable marker for monitoring P. fluorescens 10586s FAC510 in liquid culture and soil, since detection by luminometry provided a sensitive, rapid, and non-extractive technique for measurement of microbial activity. In the presence of C. steinii, bacterial activity increased, whilst predation by Cercomonas sp. caused a reduction. The different responses of the bacterial inoculum may reflect the divergent balance between bacterial turnover, leading to nutrient regeneration, and grazing strategies, which are selective of the two protozoa. The distribution of bacterial cells in the soil pore network was manipulated by adjustment of the antecedant matric potential prior to inoculation. The ability to predominantly place bacterial and protozan cells in specific pore size classes was confirmed by partial chloroform fumigation and resin impregnated sections of soil.

Helicoverpa armigera (Hübner) is an economically important pest species, affecting many crops worldwide. It has a history of quickly developing resistance to numerous chemical sprays. A closely related native Australian species, H. punctigera Wallengren, is also proving to be a pest of economic interest in agriculture. The implementation of genetically modified insecticidal cotton resulted in a massive reduction in chemical control as transgenic crops expressing Bt toxins are more effective at controlling insect pests than chemical sprays. Bt cotton also provided a challenge to resistance management - how to best prevent Helicoverpa spp. acquiring resistance to Bt, when most of the cotton planted contains Bt toxins which are selecting for resistance. The Australian cotton industry deploys mandatory refuges on farms that grow Bt cotton to generate susceptible moths that can interbreed with any resistant partners to dilute resistance. However, while significant work has been undertaken to determine their efficacy, it is not understood whether modifying specific aspects of refuges might impact on the evolutionary processes which result in resistance. This thesis investigates refuge management strategies to maximise production of Bt-susceptible Helicoverpa spp. moths. As part of this, experiments were undertaken to determine if refuges could be made more attractive (for oviposition) and productive (in terms of the number of larvae surviving to adulthood). As part of this, the invertebrate communities of pigeon pea and cotton refuges were compared and manipulated. Over three Australian summers (2010/11 to 2012/13), field experiments aimed to determine if there was a maximum carrying capacity of H. armigera in refuges and whether this upper density varied between two common refuge types – non-Bt cotton and pigeon pea. Varying densities of H. armigera were added to plants and their survival observed. Measurements of plant characteristics, wild Helicoverpa spp. moth oviposition rates and invertebrate communities were also taken. An insecticide treatment to which H. armigera was resistant was incorporated in the final two seasons to determine if densities of H. armigera might be increased by removing predators and interspecific competitors from the system.

This thesis analyses the development of safety regulation for the intentional release of genetically modified organisms (GMOs) between 19 89-92, especially in Britain in its European context, and by contrast to the USA. The thesis emphasizes the practical dilemmas of GMO regulation in accommodating uncertainties about public unease and environmental harm. It serves as a case study of safety regulation as a constructed rationality, of national regulatory styles, and of environmental precaution. In anticipating hazards prior to evidence of harm, GMO regulation had a contested 'rational' basis. Regulators encountered disputes in defining the risk problem, in establishing risk-management institutions, and in reducing scientific uncertainty about potential harm. Insofar as GMO regulation had a precautionary content, it undermined the 'rational' stereotype of risk-assessment steps. Both the precautionary potential and its limits derived from the project of overcoming obstacles to a biotechnology market. This meant symbolically normalizing GMOs as benign products, while specifying testable ecological uncertainties rooted in some naturalistic analogy. Technical 'risk' abstracted potential harm from issues of socioagronomic control which underlay the earlier environmental controversy. The thesis argues for recasting theoretical models of safety regulation as a 'technical' or 'procedural' rationality. GMO regulation contained poles of tension which such theoretical models attribute to antagonistic rationalities. Broadly speaking, the regulatory system was managing an internal contradiction between social legitimacy and commercialization. The difficulties of GMO regulation arose from its implicit role in legitimizing biotechnology, by default of any democratic procedure for adjudicating a contentious technoscientific development.

A luminescence based marker system was developed for detection of genetically modified Pseudomonas fluorescens and E. coli. During batch growth in liquid culture, luminescence measured by luminometry was directly proportional to biomass concentration and enabled detection of 104 - 106 cells ml-1 of P. fluorescens and 101 cells ml-1 of E. coli, in actively growing cultures. Following inoculation into soil, detection levels were reduced ten-fold. After the subsequent utilisation of available nutrients, activity and luminescence decreased and luminometry then provided a direct, non-extractive means of measuring population activity of lux-marked inocula. Potential luminescence, measured as luminescence following amendment with nutrients, enabled assessment of the rate of reactivation of the lux-marked inocula and quantification of the size of the activatable population. Both these techniques, and traditional techniques, were used to investigate the survival of P. fluorescens and E. coli in soil microcosms. The effect of matric potential and indigenous organisms on luminescence and on survival of P. fluorescens was assessed. Matric potential significantly decreased the activity of both introduced and indigenous populations, but the indigenous population also significantly decreased the activity and biomass concentration of the introduced P. fluorescens population. Use of luminometry as a non-extractive measure of biomass concentration provided qualitative correlation with viable cell concentration, suggesting its potential for rapid enumeration of marked inocula. Reactivation of cells at increased matric stress was decreased, but use of high substrate:cell ratios at -30 kPa produced higher levels of luminescence and may, therefore, improve the use of luminometry as an estimate of biomass.

La introducció de noves tecnologies en l'àmbit agroalimentari ha revolucionat la eficiència pel que fa al sector productiu, si bé també ha tingut efectes rellevants pel que fa a la demanda, que cal analitzar amb profunditat. Aquest és el cas dels productes modificats genèticament, que solen ser productes de consum habitual, per la qual cosa han creat una important polèmica.
Hi ha diferents agents del mercat que participen en aquest debat, com és el cas dels productors, els quals en poden obtenir benefici pel que fa a millores en eficiència i reducció de costos. Tanmateix, és essencial tenir en compte l'opinió dels consumidors alhora d'analitzar els límits comercials de les noves tecnologies.
Per a conèixer l'opinió dels consumidors cal entendre el seu procés d'elecció i de valoració dels productes agroalimentaris, com també el seu comportament a l'hora de prendre decisions en el mercat alimentari.
Aquesta tesis comença amb una extensa revisió bibliogràfica en relació a les actituds dels consumidors vers els productes modificats genèticament.
Seguidament s'analitza la intenció de compra dels consumidors utilitzant experiments d'elecció. L'objectiu principal d'aquesta tesi s'ha assolit amb els subsegüents capítols on s'han analitzat els diferents elements cognitius que porten a l'individu a la intenció final de compra. La metodologia emprada ha estat principalment equacions estructurals i experiments d'elecció.
S'han emprat diferents bases de dades per a poder contrastar totes les hipòtesis plantejades en l'estudi.
La principal contribució d'aquesta tesi ha estat determinar quins son els elements que cal considerar per entendre el procés de decisió del consumidors vers els aliments modificats genèticament, com també determinar que existeix un clar escepticisme en la societat pel que fa a aquests productes. Les conclusions d'aquesta tesi posen en manifest la necessitat de una millor estratègia de comunicació que permeti als consumidors una percepció de millor i major qualitat d'informació en relació a aquests productes.
The introduction of new technologies in food production not only has revolutioned its productive efficiency but has exerted important demand side effects that cannot be dismissed and require careful analysis. This is especially the case of genetically modified (GM) food, typical daily consumed products (e.g. GM milk, tomato, maize, etc) given its considerable interest and worldwide public controversy. Several stakeholders are place, on the one hand, farmers and manufactures perceive potential benefits from efficiency improvements as well as costs, mainly resulting from the need to reimburse intellectual property rights for new foods. From a demand side, GM food brings new products in the food chain and ultimately fulfils to an extent a latent consumer's preference for diversity. Hence, in order to determine the limits of technology dissemination and transfer it becomes a key issue to examine and disentangle which demand side factors stand behind technology acceptance. This calls for a better understanding of consumer choice, its valuation and the behavioural decision making process.

The valuation of a new technology good implies the provision of information from several sources - public and private, formal and informal, etc - while conditioning on the credibility and trustworthiness of each relevant information source. Given the information available, a further issue concerns attitude expression and formation, which ultimately leads to the final question regarding product valuation and consumer preference. Interestingly, attitude formation for the case of new food generations is found to be explained by a combination of risk and benefit perceptions, based on different elements such as a general attitude towards science, knowledge, trust, education and values, among others.

This thesis begins with an extensive review of the literature on attitudes towards GM agro-food products- chapter 2- by means of a literature review. Next it conducts a study on consumers' final intentions towards agro - food products, entailing the application of choice experiments - chapter 3. moreover, the main core of this dissertation is aiming at examining what gives rise to the final choice (chapters 4 to 6). The methodologically employed relies on structural equation modelling, although in each chapter different databases have been used. The relative strengths of each database allow us taking into account the information required to test specific hypotheses. Finally, some conclusions are drawn in chapter 7 which summarises the main findings of each chapter and put them in context of the main discussion questions examined in this thesis.

The main contribution of this thesis has been to point out a set of features that condition choice, intentions and revealed purchase intentions regarding GM food. Our wok is innovative in that: 1) it contains an innovative literature review; 2) develops choice modelling of scenarios that includes, apart from methodological features, the choice between GM processed and organic food; and 3) unlike most of behavioural analysis it exploits using structural equation modelling, several theoretical structures that explain decision making and, particularly, the role of broader attitudes towards science and technology in influencing purchase intentions alongside the effect of both risks attitudes and risk perceptions in determining consumer acceptance.

The sequence and findings of the thesis can be summarized as follows: drawing from an exhaustive review of published finding regarding public intentions towards GM food we conclude that there is consumer's scepticism explaining a negative valuation towards GM food. However, the most important outcome has been to find that final purchasing intention is the result of a complex decision process which only has been partially explained in the literature. Therefore, upon the definition of a general theoretical model this thesis has tried to explain in the different chapters some specific issues of such model as well as to validate it in a croos - country exercise in the last chapter. Results from this study rises a key policy implication: the need of a well defined communication stralegy to provide information in such a way that allows individuals to feel adequately informed.

Els cultius Modificats Genèticament (MG) podrien tenir efectes negatius sobre el medi ambient, és per això que a la Unió Europea és obligatori el seguiment post-comercialització a llarg plaç, anomenat Seguiment General (SG), “General Surveillance-GS” en anglès. Malgrat aquesta obligatorietat, el SG que s’està duent a terme actualment no té la capacitat necessària per a detectar els efectes ambientals dels cultius MG. Per això, l’objectiu de la tesi és millorar la capacitat de detecció del SG a través de: a) la selecció d’artròpodes no-diana adients per al seguiment del panís MG, i b) identificar les xarxes de seguiment ambiental existents en el territori i determinar si les dades que generen tindrien la sensibilitat necessària per a detectar efectes ambientals del maneig agrícola a llarg plaç, com per exemple del cultiu del panís MG. Dos dels tàxons més adients per al seguiment dels possibles efectes ambientals del panís MG són les papallones (Lepidoptera:Papilionoidea) i els caràbids (Coleoptera:Carabidae), ja que podrien ser molt sensibles tant als impactes dels panissos resistents a insectes (Bt) com als tolerants a herbicides (HT). Per a determinar l’abundància, la variabilitat i la distribució de papallones i caràbids en l’agroecosistema del panís, durant dos anys es van mostrejar diferents hàbitats i localitats de tres zones de cultiu de panís al nordest de la Península Ibérica. Les papallones es van mostrejar mitjançant recomptes visuals i van ser abundants en tots els hàbitats mostrejats. La majoria de les espècies observades estarien exposades als efectes del cultiu del panís MG perquè les plantes larvàries eren presents en tots els hàbitats, incloent els camps de panís. Es va desenvolupar i aplicar un sistema per a seleccionar les papallones més adients per al seguiment dels impactes del panís MG. Les espècies més adients depenien de la regió geogràfica, però els indicadors que es podrien mostrejar amb menys esforç en les tres regions serien les agrupacions d’espècies i les espècies Polyommatus icarus (Rottemburg) i Pieris napi (L.). Els caràbids es van mostrejar mitjançant trampes de gravetat i van ser més abundants en els marges dels camps de panís. L’espècie indicadora més adient va ser l’omnívor Pseudoophonus rufipes (De Geer), ja que va satisfer els criteris d’abundància, rellevància, sensibilitat i facilitat de mostreig. Seria recomanable realitzar el seguiment del grup de caràbids carnívors com a indicadors de biodiversitat i control biològic d’invertebrats. La xarxa de seguiment ambiental més adient va ser la xarxa de seguiment de les papallones de Catalunya (CBMS). Es van analitzar dades obtingudes de 29 punts de mostratge durant un període de 15 anys. Aleshores es va determinar la capacitat de la prova-t de Welch per a detectar diferències en l’abundància de 12 indicadores en dos tipus de paisatge. La capacitat de detecció va ser molt bona per a alguns indicadors (es podria detectar un canvi poblacional del 30%). La capacitat de detecció depenía del tamany mostral i de la variabilitat i augmentava ràpidament quan s’agregàven espècies i anys de dades. La conclusió general d’aquesta tesi és que el SG dels panissos MG es podria millorar considerablement a través del mostratge de papallones i caràbids. A més, les dades generades per les xarxes de seguiment de papallones permetrien augmentar força la capacitat de detecció d’efectes ambientals.
El cultivo de plantas Modificadas Genéticamente (MG) podría tener efectos negativos sobre el medio ambiente. Por esta razón, en la Unión Europea es obligatorio el seguimiento post-comercialización de estos cultivos, llamado Seguimiento General (SG) o “General Surveillance-GS” en inglés. Sin embargo, en su forma actual, el GS carece de la capacidad necesaria para la detección de efectos ambientales de los cultivos MG. Así, el objetivo de esta tesis es potenciar la capacidad de detección del SG mediante: a) la selección de artrópodos no-diana que sean buenos indicadores para el seguimiento del maíz MG, y b) el análisis de datos generados por redes de seguimiento ambiental para determinar si tendrían la sensibilidad necessaria para detectar efectos ambientales derivados de prácticas agronómicas o de los cultivos MG. La literatura indica que dos de los taxones más adecuados para el seguimiento de los posibles efectos ambientales del maíz MG son las mariposas (Lepidoptera: Papilionoidea) y los carábidos (Coleoptera: Carabidae), ya que pueden ser sensibles a los impactos de maíces resistentes a insectos (Bt) y tolerantes a herbicidas (HT). Durante dos años se muestrearon diferentes hábitats y localidades en tres zonas de cultivo maicero en el nordeste de la Península Ibérica para determinar la abundancia, variabilidad y distribución de las mariposas y carábidos en el agroecosistema del maíz. Las mariposas se muestrearon mediante recuentos visuales y fueron abundantes en todos los hábitats muestreados. La mayoría de las especies observadas podrían estar expuestas a los efectos del cultivo del maíz MG ya que se encontraron plantas nutricias de las larvas en todos los hábitats, incluidos los campos de maíz. A partir de los resultados de campo y de la literatura, se desarrolló y aplicó un sistema para seleccionar las mariposas más adecuadas para el seguimiento de los impactos del maíz MG. Las mejores indicadoras dependieron fuertemente de la región considerada, sin embargo, en este estudio las indicadoras que se podrían muestrear con el menor esfuerzo fueron los indicadores multiespecie, Polyommatus icarus (Rottemburg) y Pieris napi (L.). Los carábidos se muestrearon mediante trampas de gravedad y fueron más abundantes en los márgenes de los campos de maíz por lo que éste sería la mejor localización para el seguimiento. El mejor indicador fue el omnívoro Pseudoophonus rufipes (De Geer), cumpliendo los criterios de abundancia, relevancia, sensibilidad y facilidad de muestreo. También se recomienda el seguimiento del grupo de carábidos carnívoros como indicador de biodiversidad y de control biológico de invertebrados. La red de seguimiento ambiental que podría utilizarse con mayor facilidad es la red de seguimiento de mariposas de Cataluña (CBMS). Se analizaron los datos obtenidos en 29 puntos de muestreo durante un período de 15 años. Se determinó la capacidad de la prueba-t de Welch para detectar diferencias en la abundancia de 12 indicadores en dos tipos de paisaje. La capacidad de detección fue muy buena para algunos indicadores (pudiéndose detectar un cambio poblacional del 30%). La capacidad de detección dependía del tamaño muestral y de la variabilidad, y crecía rápidamente a medida que se agrupaban especies y años de datos. La conclusión principal de la tesis es que el SG de los maíces MG se podría mejorar considerablemente a través del muestreo de determinadas mariposas y carábidos. Asimismo, se podrían utilizar los datos generados por las redes de seguimiento de mariposas ya que permitirían incrementar la capacidad de detección de efectos ambientales sin incurrir en elevados costes.
Genetically Modified (GM) crops could adversely affect the environment, therefore long-term post-market monitoring (General Surveillance, GS) is mandatory in the EU. However, in its current form, GS lacks the required sensitivity to detect environmental effects of GMs. The aim of this thesis is to increase the effect detection capacity of GS through a) selection of suitable non-target arthropods (NTAs) to monitor GM maize, and b) identify suitable environmental surveillance networks (ESNs) and determine if their data is sufficiently sensitive to detect long term environmental effects of agricultural practices –such as GM cultivation. Butterflies (Lepidoptera: Papilionoidea) and carabids (Coleoptera:Carabidae) were identified as the most appropriate surrogate taxa for monitoring effects of GM maize on non-target organisms due to their potential sensitivity to impacts of GM maize expressing insect resistance (Bt) or herbicide tolerance (HT). A two-year field survey was carried out in different maize agroecosystems in the north-east of the Iberian Peninsula to determine abundance, variability and distribution of carabids and butterflies across different habitats and sites. Butterfly adults were sampled by transect-counts and they were abundant in all habitats sampled. Most species could be exposed to effects of GM maize because their larval host plants were present in maize fields and neighbouring habitats. A step-by-step selection procedure was developed and applied, finding that the most appropriate species for monitoring GM effects depended on the region considered. Of these, the indicators requiring the lowest sampling effort were the multispecies pools and the single species Polyommatus icarus (Rottemburg) and Pieris napi (L.). Carabid adults were sampled by pitfall trapping and they most abundant in field margins. The best indicator was the omnivore Pseudoophonus rufipes (De Geer), satisfying criteria of abundance, relevance, sensitivity and ease of sampling. In addition, the carnivore group was a good indicator of biodiversity and invertebrate biological control. The most suitable environmental surveillance network (ESN) in the study region was the Catalan Butterfly Monitoring Scheme (CBMS). A 15-year dataset from 29 recording sites was analysed to determine the capacity of Welch’s t-test to detect differences in abundance of 12 farmland butterfly indicators across land-use types. Detection capacity was very good (a change below 30% could be detected) for some indicators and it depended mainly on sample size and variability of the data. Detection capacity rapidly improved when species were aggregated into multispecies indicators and when the longer time periods were used. The main conclusion of this thesis is that GS of GM maizes could be substantially improved through field sampling of butterflies and carabids; and that data from butterfly monitoring schemes would allow to greatly increase effect detection capacity.

Thesis (M.S.)--School Computing and Engineering. University of Missouri--Kansas City, 2004.
"A thesis in mechanical engineering." Typescript. Advisor: Bryan R. Becker. Vita. Title from "catalog record" of the print edition Description based on contents viewed Feb. 28, 2006. Includes bibliographical references (leaves 71-73). Online version of the print edition.

Quite a number of important crops have been genetically modified with genes for agronomically important traits, such as insect and viral resistance. As the numbers of genetically modified foods continue to increase on the market, the need for rapid development of GMO detection methods is indispensable. This study was carried out to detect if genetically modified potatoes exist on food market in Turkey. Thirty samples from different places were collected. Using a DNA based PCR method, potato samples were examined for the presence of 35S promoter, Nos terminator, neomycin phosphotransferase (nptII) genes, and synthetic cry3A gene which is the general transgene in all approved Newleaf transgenic potato lines. The experimental design of this study was to detect Newleaf insect resistant lines. In 11 samples at least one genetic element was detected. Sample R from Ankara has shown to be belonging to Newleaf insect resistant lines. Since 35S promoter was not detected in samples M3, 14 and F1, it is proposed that they are belonging to Newleaf virus and insect resistant lines (Newleaf plus or Newleaf Y). Although Nos terminator was not detected in samples H2, Z2 and D, cry3A fragments amplified in those samples have been verified that they are from the synthetic cry3A regions of Newleaf lines. The detected synthetic cry3A gene in GM potatoes was amplified by specific primers, which cannot amplify Bacillus thuringiensis tenebrionis natural cry3A gene. In addition, the authenticity of the synthetic cry3A PCR products were confirmed by both sequencing and restriction digestions. Our results showed that genetically modified Newleaf potatoes exist in food market in Turkey. Further studies by accredited laboratories are strongly recommended.

This paper attempts to present and analyze problems that may arise from the use of Genetically Modified (GM) products and issues raised by the Intellectual Property (IP) rights that Genetic Engineering (GE) companies have on their products. Arguments in favor and against the existence of health risks and environmental risks of GM products are presented. The European policy of the socioeconomic effects of the GM products is discussed and is proposed the application of the precautionary principle for the prevention of unintended consequences from the GM products to other than health and environmental domains. The need of IP rights is supported, but is also suggested an IP rights flexibility. Do IP rights violate the rights of all people to a nourishing life, natural resources, the right to decide about what they eat and the right to live in a viable ecosystem? Finally, I provide an analysis of the effects on the farmers due to the IP rights on GM crop for cultivation and state dependency issues that may occur.

Cellulose is the major biopolymer on earth with tremendous economic importance as it has been utilised in a multitude of industrial applications including tissueengineering products, composite materials and electronics. It exhibits outstanding physical and mechanical properties when compared to plant-based cellulose. Although A. xylinum is the most efficient producer of bacterial cellulose (BC), its long doubling time results in insufficient yields and high cost. In this study, a novel and functional BC production system was developed by recombinant DNA technology. The simultaneous expression of bacterial cellulose synthase operon (bcsABCD) and its upstream operon (cmcax and ccpAx) was achieved by pBCS and pCDF, respectively. Three different Escherichia coli strains were utilised as host microorganisms: E. coli BL21 (DE3), E. coli HMS174 (DE3) and E. coli C41 (DE3). It was verified that bcsABCD and the upstream operon were successfully cloned and expressed in E. coli strains. Fermentation of genetically modified (GM) strains was conducted at various IPTG concentrations (0.025, 0.05, 0.1, 0.2, 0.5 and 1.0 mM) and various temperatures (22, 30, and 37 °C). BC production was achieved by genetically modified E. coli HMS174 (DE3) in the presence of 0.025 mM IPTG at 22°C. GM E. coli C41 (DE3) accomplished the production when IPTG supplement was lower than 0.2 mM at 22°C or 30°C. The products were characterised by SEM and FTIR, which exhibited that morphology of product was stain-specific. Finally a dynamic mathematical model was developed to design a fed-batch system capturing characteristics incorporating acetate inhibition and cell death, which allowed predicting glucose consumption, acetate production and induction time for batch cultures, resulted in a volumetric productivity of 1.7 mg/L.h. In conclusion, this thesis reports the development of a novel BC production system by creating valuable cellulose-producing E. coli strains, resulting in a reproducible and stable recombinant expression system for potential improvement of BC.

Pulmonary fibrosis is a feature of a number of important lung diseases, the commonest of which is idiopathic pulmonary fibrosis (IPF). IPF is refractory to current treatments and has a poor prognosis, so development of novel therapeutic strategies is paramount. Injury to the alveolar epithelium plays a central role in the pathogenesis of IPF, and represents a logical therapeutic target. Keratinocyte growth factor (KGF) exerts protective effects on the alveolar epithelium and is a potential therapeutic candidate. There is growing interest in combined gene and cell therapy approaches, and in this study the therapeutic potential of macrophages genetically modified to express KGF was explored in the mouse bleomycin model of pulmonary fibrosis. Lentiviral vectors were generated expressing KGF, and used to transduce murine BMDM and the IC-21 murine macrophage cell line. KGF-transduced IC-21 cells were characterised and were shown to induce proliferation in mouse lung epithelial cells in vitro. A luciferase-expressing lentiviral vector was produced, and IC-21 cell were transduced to express luciferase. Luciferase-transduced IC-21 cells were tracked in vivo using bioluminescence imaging after delivery to the lungs of mice, and were retained in the lungs of bleomycin-treated mice, but not saline-treated controls, and localised to injured areas. KGF-transduced IC-21 cells were delivered to mice during both the inflammatory and fibrotic phases of the mouse bleomycin model, and no improvements in outcomes reflecting alveolar-capillary membrane permeability, inflammation, cytotoxicity and fibrosis were demonstrated. The reasons for the lack of therapeutic efficacy of KGF-transduced macrophages were unclear, but delivery of macrophages per se was associated with an increase in inflammatory mediators. In conclusion, exogenously delivered macrophages were shown to localise to sites of lung injury, but KGF-transduced macrophages were not found to have therapeutic efficacy in bleomycin-treated mice. Future work should better characterise the effects of exogenous macrophage delivery in pulmonary fibrosis.

This research aims to analyze people’s perceptions about the potential risks associated with the presence of genetically modified organisms (GMOs) in food products. We formulated research questions and hypotheses based on parameters, including age, gender, state of residence, and more to analyze these perceptions. We conducted an online nationwide survey across the United States and recruited participants from the general population to understand their perceptions about risks for GMOs and GM foods. We formulated a set of questions regarding the effects of GMOs on food products (including both the pre- and post-study questions) and investigated the changes in people’s perceptions after reading selected news releases about GMOs. The survey responses were collected and categorized according to the research parameters and statistical assessments were conducted to test the hypotheses. Additionally, we introduced a novel approach to analyze the responses by creating a mind-map framework for both the pre- and post-study responses. We found that people residing in the southern region of the United States responded more positively toward GMOs when compared to individuals residing in the northeast, west and mid-west regions. We also deduced that people’s perceptions about GMOs were not significantly different from each other whether they resided in states with Republican or Democrat/non-partisan party affiliations. Further, we observed that the male participants responded more negatively compared to the female participants across the nation. We compared the results obtained from respondents in the general population to those from a group of Computer Science students at North Dakota State University who completed the same survey. We found that students considered GMOs less risky when compared to the general population. A third research study compared participants from the general population to a group of participants who were recruited from the general population. The second group didn’t read the news releases that separated the survey’s pre- and post-study questions. We observed that the news releases impacted the participants from the first group and, eventually, changed the individuals’ perceptions about GMOs when compared to the participants from the second group who possessed no or fewer perception changes.