In 1991, India adopted neoliberalism, a system of political economic practices that promotes private property and free trade, as its political and economic system to promote development in their country. India's neoliberal reform has created issues surrounding human development, resource accumulation, and power struggles. Eleven years later, in 2002, Bt cotton was introduced to the Indian agricultural sector. This research examines how the genetically modified organism Bt cotton is being used to commodify nature in the context of agriculture under neoliberalism. The research focuses on the dispossession of the rural farmers through the commodification of agriculture using Bt cotton. Dispossession of the rural farmers happen through the implications that arise from the commodification of nature. Through Marxist theory of primitive accumulation, this research analyzes accumulation by dispossession and how it neglects the working class and its struggle in rural India. Through this examination, the research will argue alternatives to the dispossession of the working class and the commodification of nature through Bt cotton. Dispossession, in this research, is examined both through working class, but also through the dispossession of biodiversity. Through the loss of biodiversity, the rural farmers are becoming dispossessed from a more sustainable environment. Along with these goals, the research will also incorporate themes of food security through changing landscape of agriculture due to the incorporation of Bt cotton. This research argues the contradictions that are presented through the commodification of agriculture under neoliberalism and provide a contribution to social justice literature, and our understanding of the relationship between technology and the commodification of nature.

Synthetic biology enables the engineering of interactions between living medicines to overcome the specific limitations of monotherapies. A major challenge facing tumor-antigen targeting therapies like chimeric antigen receptor (CAR)-T cells is the identification of suitable targets that are specifically and uniformly expressed on heterogeneous solid tumors. In contrast, certain strains of bacteria are gaining recognition as a new class of antigen-agnostic cell therapy due to their selective growth within the immunosuppressive niche of the solid tumor microenvironment (TME). In response, this dissertation aims to pair the cytotoxicity of CAR-T cells with the antigen-independent specificity of tumor-colonizing bacteria to create a new strategy for solid tumor recognition. Here, we reprogram the probiotic strain of E. coli Nissle 1917 to release synthetic CAR targets and human chemokines directly within the solid tumor core. To enable universal targeting, we design synthetic targets to bind ubiquitous components of the TME and broadly tag tumor tissue for CAR-mediated lysis. We demonstrate that these targets robustly coat the surface of cancer cell lines and lead to effective killing by CAR-T cells across various cancer types. We additionally show that injected probiotics selectively grow within the tumor core and maintain target production ¬ in situ – leading to therapeutic efficacy across multiple genetically distinct tumor models. Within this dissertation, we also reveal that intratumoral bacteria provide natural adjuvant effects that serve to activate and increase the effector functions of CAR-T cells in vivo. However, we discover that this can lead to early T cell exhaustion and terminal effector differentiation. To mitigate the counterproductive effects of overstimulation, we generate a new probiotic strain with reduced inflammatory properties that significantly improves CAR-T cell phenotype – leading to enhanced therapeutic benefit in a human model of leukemia. We conclude by discussing the numerous avenues available to optimize cross-Kingdom signaling and to ultimately leverage the full therapeutic benefit of combined cell therapies for future translation. Altogether, this dissertation highlights the potential of the probiotic-guided CAR-T cell (ProCAR) platform to address the critical roadblock of identifying suitable CAR targets by providing an antigen in situ that is orthogonal to both healthy tissue and tumor genetics – and, in turn, aims to establish the foundation for engineered communities of living medicines.

No description available.

This study represents the behavioural component of a larger project investigating the life history tactics, physiological resource allocation and behavioural time budgeting of a genetically engineered animal. The "supermouse" is a transgenic strain (mMT-1/rGH) that has one chromosome genetically engineered with extra copies of rat growth hormone genes, each fused to a metallothionein-1 promoter. The GH transgenes are permanently incorporated into the genome of the mouse and are inherited as a block, in a Mendelian manner. "Supermice" exhibit an accelerated growth rate and reach body weights twice that of their normal siblings: both transgenic mice and normal mice are obtained by crossing transgenic males to normal females. Although there must be increased costs associated with achieving their higher growth rate, these: mice show no increases in their specific feeding rates. Consequently there must be a reallocation of resources among various physiological and behavioural demands. The reality of such tradeoffs is known as the Principle of Allocation and predicts that reductions in behavioural activities might be one avenue for realizing extra growth. To test this, six components of the behavioural time budget (resting, locomotion, wheel running, feeding, drinking and grooming) were compared between transgenic and normal mice. Infra-red videocameras recorded the activities of individual male mice in artificial enclosures over 24 hours. The time spent in each bout of activity was recorded and compared. Transgenic mice out-slept their normal counterparts by 126% (an increase of 3.4 h) and were only 53.83% as active in terms of locomotion and wheel running as normal mice. Pooling the data revealed that on average, large mice spent more time at rest and less time engaged in locomotion. Slight but significant decreases in time spent drinking and grooming were also found. Transgenic mice spent only 77.01% as much time drinking, and 69.01% as much time grooming as normal mice. No difference in the amount of time spent feeding was found. / Thesis / Master of Science (MS)

A series of experiments were used to evaluate three culture media and two incubation temperatures for their ability to support development of DNA microinjected porcine zygotes. Development in vitro was compared between embryos collected from postpubertal and prepubertal donors and between embryos injected with DNA into the pronucleus and the cytoplasm. Additionally, embryos were analyzed by the polymerase chain reaction (PCR) for the presence of the transgene. One-cell embryos (n=458) were recovered from 36 postpubertal gilts in Experiment 1. Injected and control embryos were cultured in modified media NCSU-23 (mNCSU-23), NCSU-37 (mNCSU-37), and CZB at 37°C and 38.8°C for 7 d. In Experiment 2, one-cell embryos (n=245) were collected from postpubertal (n=15) and prepubertal (n=14) gilts, microinjected with DNA, and cultured in medium mNCSU-23. Superovulated prepubertal gilts (n=22) were flushed in Experiment 3 to yield 343 one-cell embryos which had DNA injected into the cytoplasm or pronucleus. Whole embryos were assessed by PCR. Mean percentages of embryos developing to the expanded or hatched blastocyst stage in mNCSU-23 and mNCSU-37 did not differ from each other (p>.05), but both were greater than the development in CZB (p<.05). Development was greater at 38.8°C (p<.05) than at 37° C. Microinjection of DNA decreased the developmental percentage (p<.05) from that of non-injected controls. Embryos collected from postpubertal gilts had a higher percentage (68.0 ± 3.4) of expanded and hatched blastocysts than embryos from prepubertal donors (29.0 ± 4.6, p<.05). No difference was seen in development between embryos injected in the pronucleus or cytoplasm (p>. 05), but development for both was less than for control embryos (p<.05). Results of PCR analysis indicated that 40% of the embryos developing to the expanded blastocyst stage were positive for the transgene compared to a rate of 60% positive for degenerate embryos. These studies show that DNA microinjected porcine zygotes can be cultured to the expanded blastocyst stage in media mNCSU-23 and mNCSU-37 at 38.8°C. Microinjection of DNA decreases survival of embryos collected from both postpubertal and prepubertal sources, but postpubertal embryos exhibit a higher rate of development. Cytoplasmic injection does not improve embryo viability in vitro above that of pronuclear injection. Finally, whole embryo analysis by PCR is possible, but cross specificity of human Protein C and whey acidic protein (WAP) oligonucleotides for endogenous porcine DNA is strong and creates difficulty in applying PCR analysis to embryos microinjected with WAP-PC transgenes. / Master of Science

Aquifers are an important storage location and source of fresh groundwater. They may become polluted by a number of contaminants including mobile divalent radionuclides such as strontium-90 which is a byproduct of uranium fission. A method for remediating such divalent radionuclides is sequestration through co-precipitation into calcium carbonate. Calcium carbonate precipitation occurs naturally but can be enhanced by the use of ureolytic microorganisms living within the aquifer. The microbial enzyme urease cleaves ammonia from urea (added as a stimulant to the aquifer) increasing the pH and subsequently pushing the bicarbonate equilibrium towards precipitation. Laboratory experimentation is necessary to better predict field scale outcomes of remediation that is driven by ureolytic calcium carbonate co-precipitation. To aid in such laboratory experiments, I constructed two ureolytic organisms which contain green fluorescent protein (GFP) so that the location of the microbes in relation to media flow paths and precipitation can be viewed by microscopy in a 2- dimensional porous medium flow cell reactor. The reactor was operated with a parallel flow regime where the two influent media would not promote microbially induced calcium carbonate precipitation until they were mixed in the flow cell. A demonstration study compared the results of parallel flow and mixing in the reactor operated with and without one of the GFP-containing ureolytic organisms. The growth and precipitation of calcium carbonate within the reactor pore space altered flow paths to promote a wider mixing zone and a more widely distributed overall calcium carbonate precipitation pattern. This study will allow optimization of remediation efforts of contaminants such as strontium-90 in aquifers. / Graduation date: 2012

A obtenção de plantas geneticamente modificadas tornou-se uma ferramenta biotecnológica de grande valor, contribuindo nos programas de melhoramento. Plantas transgênicas contendo genes de peptídeos antibacterianos são uma boa alternativa nos programas visando resistência às bactérias. A cecropina é um peptídeo isolado de inseto que apresenta ação antibacteriana contra diversas bactérias, entre elas, a Xylella fastidiosa causadora da clorose variegada dos citros (CVC). Com isso, este trabalho teve dois objetivos: 1) obter plantas transgênicas dos cultivares copa de laranja &#39;Hamlin&#39; e laranja &#39;Pêra&#39; (Citrus sinensis L. Osbeck) com o gene cecropin MB39 dirigido pelos promotores do gene da fenilalanina amônia-liase (PAL) clonado de citros (CsPP) e de Arabidopsis thaliana (AtPP), que conferem expressão gênica nos vasos do xilema. Obter plantas transgênicas de laranja &#39;Hamlin&#39; e laranja &#39;Valência&#39; com o gene GUS dirigido pelo promotor AtPP. 2) avaliar plantas de laranja &#39;Valência&#39; transformadas com o gene cecropin MB39 inoculadas com Xylella fastidiosa. As transformações genéticas foram realizadas através do co-cultivo de explantes, coletados de plantas cultivadas in vitro (segmentos de epicótilo) ou em casa de vegetação (segmentos internodais), com Agrobacterium tumefaciens. Para isso, utilizou-se a estirpe EHA-105 de A. tumefaciens contendo os vetores binários CsPPCEC/2201, AtPPCEC/2201 ou AtPP-GUS/2201. A seleção das gemas transformadas foi feita em meio de cultura contendo o antibiótico canamicina, e para confirmação da transformação foram realizados teste histoquímico GUS e análises moleculares de PCR e &#39;Southern blot&#39;. As gemas regeneradas foram microenxertadas em citrange &#39;Carrizo&#39; ou laranja &#39;Hamlin&#39; não transgênicos. Foi possível a obtenção de gemas transformadas para todos os cultivares, porém a regeneração de plantas ocorreu para laranja &#39;Hamlin&#39; transformada com os vetores CsPPCEC/2201, AtPPCEC/2201 e AtPP-GUS/2201. Para a avaliação da resistência de plantas transgênicas a Xylella fastidiosa foram selecionadas nove plantas de laranja &#39;Valência&#39; transformadas com o gene cecropin MB39, sendo que em quatros plantas o gene está sendo dirigido pelo promotor CsPP e nas outras cinco plantas pelo promotor CaMV 35S. Essas plantas foram multiplicadas por borbulhia e inoculadas mecanicamente com Xylella fastidiosa. Foram realizados isolamentos primários e quantificações bacterianas aos quatro e dez meses após a inoculação da bactéria para verificar a eficiência da inoculação e movimentação sistêmica da bactéria. Avaliações sintomáticas foliares também foram feitas aos oito meses após a inoculação. Os resultados dos isolamentos aos quatro meses mostraram que a inoculação da bactéria foi eficiente, já que houve crescimento bacteriano na maioria das placas com meio de cultura. A movimentação sistêmica da bactéria pôde ser detectada no isolamento aos dez meses. Das nove plantas avaliadas, uma planta transgênica apresentou crescimento populacional menor do que a da planta controle, indicando uma resistência ao patógeno. Nas avaliações sintomáticas foliares, as plantas transgênicas não diferiram do controle. / The production of genetically modified plants has become an important biotechnological tool, contributing with breeding programs. Transgenic plants expressing antibacterial peptides genes are a good alternative for breeding programs aiming to obtain bacterial resistance. Cecropin is a peptide isolated from an insect, which presents antibacterial effect against several bacteria including the citrus variegated chlorosis in which the causal agent is Xylella fastidiosa. This work had two objectives: 1) genetically transform &#39;Hamlin&#39; and &#39;Pêra&#39; sweet oranges scions with the cecropin MB39 gene under the control of the phenylalanine ammonia-lyase (PAL) gene promoter, cloned from citrus (CsPP) and from Arabidopsis thaliana (AtPP). This promoter confers gene expression within the xylem vessels. Obtain transgenic plants of &#39;Hamlin&#39; and &#39;Valência&#39; sweet oranges with the GUS gene under the control of the AtPP promoter. 2) Evaluate the resistance to Xylella fastidiosa of &#39;Valência&#39; sweet orange plants transformed with the cecropin MB39 gene. Co-culture of explants with Agrobacterium tumefaciens was used to genetically transform in vitro plants (epicotyl segments) and greenhouse plants (internodal segments). A. tumefaciens strain EHA-105 containing the CsPPCEC/2201, AtPPCEC/2201 or AtPP-GUS/2201 binary vectors were used for transformation. Culture medium containing the antibiotic kanamycin was used for selection of transformed buds. Confirmation of transgenesis was carried out by GUS assays, PCR and Southern blot analysis. Regenerated buds were in vitro grafted on non-transgenic &#39;Carrizo&#39; citrange or &#39;Hamlin&#39; sweet orange rootstocks. Transformed buds were obtained for all cultivars, however plant regeneration was only obtained for &#39;Hamlin&#39; sweet orange transformed with vectors CsPPCEC/2201, AtPPCEC/2201 and AtPP-GUS/2201. Nine &#39;Valência&#39; sweet orange plants transformed with the cecropin MB39 gene were selected for the Xylella fastidiosa resistance experiment. Four transgenic lines were controlled by the CsPP promoter and the other five lines were controlled by the CaMV 35S promoter. These transgenic lines were graft propagated and mechanically inoculated with Xylella fastidiosa. Primary isolations and bacterial quantifications were conducted on the fourth and tenth month after inoculation in order to detect inoculation efficiency and bacterial systemic movement. Leaf symptom evaluations were executed eight months after inoculations. The bacterial isolation results from the fourth month indicated that inoculations were successful, since bacterial growth could be detected in the majority of culture mediums. Systemic movement of bacteria within the plants was detected after ten months. From the nine transgenic lines tested, one presented reduced pathogen growth when compared to controls indicating resistance. Transgenic lines did not differ from controls regarding leaf symptoms.

Submitted by admin tede (tede@pucgoias.edu.br) on 2016-09-02T12:27:50Z No. of bitstreams: 1 CECY PEREIRA FIGUEIRA DA SILVA NETA MELLO.pdf: 1451626 bytes, checksum: 863ab02a5e133691994a8b778ef75e6e (MD5) / Made available in DSpace on 2016-09-02T12:27:50Z (GMT). No. of bitstreams: 1 CECY PEREIRA FIGUEIRA DA SILVA NETA MELLO.pdf: 1451626 bytes, checksum: 863ab02a5e133691994a8b778ef75e6e (MD5) Previous issue date: 2016-03-14 / With the biotechnology advancement and development, with special focus on genetic engineering, also rises the requirement of ethical fundamentals in a scenario of constant modifications. When it comes to transgenic food, there‟s controversy in the philosophical, ethical, environmental, political, legal and economic field because it‟s a subject which is inside of the daily reality, with very fast changes and in that context, the Brazilian and international legislation seems to doesn‟t follow the technology velocity and Science becomes a political and economical instrument. In this article, international law and national law related to the using of genetically modified organism are analyzed, pointing out the 11.105/05 law, considering the origin, shape and evolution of genetically modified organism, the importance of obeying the environmental law when handling and distributing products resulting of these genetic modifications to the public consumption, this law analysis, going through the environmental impacts, giving importance to the cost-benefit and the risk expressly taken by the scientific community, besides the own concept of consumer, proving the need of genetically modified food labelling and the reality of what really happens in the process of patenting acquisition in the international law and in the Brazilian law and jurisprudence decisions. / Com o avanço e o desenvolvimento da biotecnologia, com enfoque especial na engenharia genética, surge também a necessidade da inserção da ética e de seus princípios em um cenário de modificações constantes. No que tange aos alimentos transgênicos, polêmicas são levantadas no campo filosófico, ético, ambiental, político, social, jurídico e econômico, pois o tema está inserido em uma realidade do nosso dia a dia, com mudanças muito rápidas e diante deste contexto, as legislações brasileiras e internacionais parecem não acompanhar a velocidade da tecnologia e a ciência passa a ser instrumento da política e da economia. No presente trabalho são analisadas leis internacionais, legislações relativas à utilização dos organismos geneticamente modificados, ressaltando-se a Lei 11.105/05, considerando a origem, a forma e a evolução do organismo geneticamente modificado, a importância da obediência às leis de Direito Ambiental no manuseio e distribuição dos produtos oriundos destas modificações genéticas para o consumo da população, a análise desta legislação, passando pelos impactos ambientais, dando ênfase ao custo benefício e ao risco assumido expressamente pela comunidade científica, discorrendo ainda sobre o direito do consumidor, além do próprio conceito de consumidor, demonstrando também a necessidade da rotulagem dos alimentos geneticamente modificados e a forma como acontece o processo para obtenção de patentes tanto na legislação exterior quanto na legislação brasileira e as decisões jurisprudenciais.

A obtenção de plantas geneticamente modificadas tornou-se uma ferramenta biotecnológica de grande valor, contribuindo nos programas de melhoramento. Plantas transgênicas contendo genes de peptídeos antibacterianos são uma boa alternativa nos programas visando resistência às bactérias. A cecropina é um peptídeo isolado de inseto que apresenta ação antibacteriana contra diversas bactérias, entre elas, a Xylella fastidiosa causadora da clorose variegada dos citros (CVC). Com isso, este trabalho teve dois objetivos: 1) obter plantas transgênicas dos cultivares copa de laranja &#39;Hamlin&#39; e laranja &#39;Pêra&#39; (Citrus sinensis L. Osbeck) com o gene cecropin MB39 dirigido pelos promotores do gene da fenilalanina amônia-liase (PAL) clonado de citros (CsPP) e de Arabidopsis thaliana (AtPP), que conferem expressão gênica nos vasos do xilema. Obter plantas transgênicas de laranja &#39;Hamlin&#39; e laranja &#39;Valência&#39; com o gene GUS dirigido pelo promotor AtPP. 2) avaliar plantas de laranja &#39;Valência&#39; transformadas com o gene cecropin MB39 inoculadas com Xylella fastidiosa. As transformações genéticas foram realizadas através do co-cultivo de explantes, coletados de plantas cultivadas in vitro (segmentos de epicótilo) ou em casa de vegetação (segmentos internodais), com Agrobacterium tumefaciens. Para isso, utilizou-se a estirpe EHA-105 de A. tumefaciens contendo os vetores binários CsPPCEC/2201, AtPPCEC/2201 ou AtPP-GUS/2201. A seleção das gemas transformadas foi feita em meio de cultura contendo o antibiótico canamicina, e para confirmação da transformação foram realizados teste histoquímico GUS e análises moleculares de PCR e &#39;Southern blot&#39;. As gemas regeneradas foram microenxertadas em citrange &#39;Carrizo&#39; ou laranja &#39;Hamlin&#39; não transgênicos. Foi possível a obtenção de gemas transformadas para todos os cultivares, porém a regeneração de plantas ocorreu para laranja &#39;Hamlin&#39; transformada com os vetores CsPPCEC/2201, AtPPCEC/2201 e AtPP-GUS/2201. Para a avaliação da resistência de plantas transgênicas a Xylella fastidiosa foram selecionadas nove plantas de laranja &#39;Valência&#39; transformadas com o gene cecropin MB39, sendo que em quatros plantas o gene está sendo dirigido pelo promotor CsPP e nas outras cinco plantas pelo promotor CaMV 35S. Essas plantas foram multiplicadas por borbulhia e inoculadas mecanicamente com Xylella fastidiosa. Foram realizados isolamentos primários e quantificações bacterianas aos quatro e dez meses após a inoculação da bactéria para verificar a eficiência da inoculação e movimentação sistêmica da bactéria. Avaliações sintomáticas foliares também foram feitas aos oito meses após a inoculação. Os resultados dos isolamentos aos quatro meses mostraram que a inoculação da bactéria foi eficiente, já que houve crescimento bacteriano na maioria das placas com meio de cultura. A movimentação sistêmica da bactéria pôde ser detectada no isolamento aos dez meses. Das nove plantas avaliadas, uma planta transgênica apresentou crescimento populacional menor do que a da planta controle, indicando uma resistência ao patógeno. Nas avaliações sintomáticas foliares, as plantas transgênicas não diferiram do controle. / The production of genetically modified plants has become an important biotechnological tool, contributing with breeding programs. Transgenic plants expressing antibacterial peptides genes are a good alternative for breeding programs aiming to obtain bacterial resistance. Cecropin is a peptide isolated from an insect, which presents antibacterial effect against several bacteria including the citrus variegated chlorosis in which the causal agent is Xylella fastidiosa. This work had two objectives: 1) genetically transform &#39;Hamlin&#39; and &#39;Pêra&#39; sweet oranges scions with the cecropin MB39 gene under the control of the phenylalanine ammonia-lyase (PAL) gene promoter, cloned from citrus (CsPP) and from Arabidopsis thaliana (AtPP). This promoter confers gene expression within the xylem vessels. Obtain transgenic plants of &#39;Hamlin&#39; and &#39;Valência&#39; sweet oranges with the GUS gene under the control of the AtPP promoter. 2) Evaluate the resistance to Xylella fastidiosa of &#39;Valência&#39; sweet orange plants transformed with the cecropin MB39 gene. Co-culture of explants with Agrobacterium tumefaciens was used to genetically transform in vitro plants (epicotyl segments) and greenhouse plants (internodal segments). A. tumefaciens strain EHA-105 containing the CsPPCEC/2201, AtPPCEC/2201 or AtPP-GUS/2201 binary vectors were used for transformation. Culture medium containing the antibiotic kanamycin was used for selection of transformed buds. Confirmation of transgenesis was carried out by GUS assays, PCR and Southern blot analysis. Regenerated buds were in vitro grafted on non-transgenic &#39;Carrizo&#39; citrange or &#39;Hamlin&#39; sweet orange rootstocks. Transformed buds were obtained for all cultivars, however plant regeneration was only obtained for &#39;Hamlin&#39; sweet orange transformed with vectors CsPPCEC/2201, AtPPCEC/2201 and AtPP-GUS/2201. Nine &#39;Valência&#39; sweet orange plants transformed with the cecropin MB39 gene were selected for the Xylella fastidiosa resistance experiment. Four transgenic lines were controlled by the CsPP promoter and the other five lines were controlled by the CaMV 35S promoter. These transgenic lines were graft propagated and mechanically inoculated with Xylella fastidiosa. Primary isolations and bacterial quantifications were conducted on the fourth and tenth month after inoculation in order to detect inoculation efficiency and bacterial systemic movement. Leaf symptom evaluations were executed eight months after inoculations. The bacterial isolation results from the fourth month indicated that inoculations were successful, since bacterial growth could be detected in the majority of culture mediums. Systemic movement of bacteria within the plants was detected after ten months. From the nine transgenic lines tested, one presented reduced pathogen growth when compared to controls indicating resistance. Transgenic lines did not differ from controls regarding leaf symptoms.

Abstract not available