The closure of the nuclear fuel cycle is a topic of interest in the sustainability context of nuclear energy. The implication of such closure includes considerations of nuclear waste management. This originates from the fact that a closed fuel cycle requires recycling of useful materials from spent nuclear fuel and discarding of non-usable streams of the spent fuel, which are predominantly the fission products. The fission products represent the near-term concerns associated with final geological repositories for the waste stream. Long-lived fission products also contribute to the long-term concerns associated with such repository. In addition, an ultimately closed nuclear fuel cycle in which all actinides from spent nuclear fuels are incinerated will result in fission products being the only source of radiotoxicity. Hence, it is desired to develop a transmutation strategy that will achieve reduction in the inventory and radiological parameters of significant fission products within a reasonably short time. In this dissertation, a transmutation strategy involving the use of the VHTR is developed. A set of specialized metrics is developed and applied to evaluate performance characteristics. The transmutation strategy considers six major fission products: 90Sr, 93Zr, 99Tc, 129I, 135Cs and 137Cs. In this approach, the unique core features of VHTRs operating in equilibrium fuel cycle mode of 405 effective full power days are used for transmutation of the selected fission products. A 30 year irradiation period with 10 post-irradiation cooling is assumed. The strategy assumes no separation of each nuclide from its corresponding material stream in the VHTR fuel cycle. The optimum locations in the VHTR core cavity leading to maximized transmutation of each selected nuclides are determined. The fission product transmutation scenarios are simulated with MCNP and ORIGEN-S. The results indicate that the developed fission product transmutation strategy offers an excellent potential approach for the reduction of inventories and radiological parameters, particularly for long-lived fission products (93Zr, 99Tc, 129I and 135Cs). It has been determined that the in-core transmutation of relatively short-lived fission products (90Sr and 137Cs) has minimal advantage over a decay-only scenario for these nuclides. It is concluded that the developed strategy is a viable option for the reduction of radiotoxicity contributions of the selected fission products prior to their final disposal in a geological repository. Even in the cases where the transmutation advantage is minimal, it is deemed that the improvement gained, coupled with the virtual storage provided for the fission products during the irradiation period, makes the developed fission product transmutation strategy advantageous in the spent fuel management scenarios. Combined with the in-core incineration options for TRU, the developed transmutation strategy leads to potential achievability of engineering time scales in the comprehensive nuclear waste management.

Orientador: Mário Roberto Hatayde / Banca: Mirela Tinucci Costa / Banca: Maria Angélica Dias / Resumo: Sabe-se que a C. spectabilis é muito utilizada como adubação verde e para controle de nematóides, com isso espalha-se facilmente, misturando-se com o milho e outros cereais contaminando as rações. O objetivo do presente estudo foi o de avaliar os efeitos tóxicos da semente de Crotalaria spectabilis, quando trituradas e misturadas à ração de cães. Nesse experimento foram utilizados 12 cães, 6 machos e 6 fêmeas, com idades variadas, divididos em três grupos de 4 animais. Os tratamentos foram divididos por grupos, sendo que cada grupo recebeu respectivamente, 0,2, 0,4 e 0,6% de sementes de C. spectabilis, por 28 dias, que foram trituradas e adicionadas à ração. Semanalmente avaliou-se o perfil hematológico e bioquímico dos cães, além do exame físico realizado diariamente. Nos exames bioquímicos realizados, houve alteração significativa da enzima GGT. Os sinais clínicos observados foram, principalmente, alopecia periocular e alopecia difusa, além de hiperceratose nasal, espirros, tosse, vômitos esporádicos, constipação e fezes ressequidas. Para o exame histopatológico foi extraído um fragmento de fígado através de agulha tipo TRU-CUT®. Observou-se hepatócitos tumefados, vacuolização de células nos sinusóides, necrose focal, anisocitose, cariomegalia, células de Kupffer com variadas pigmentações, fibrose e presença de pigmento biliar nos hepatócitos caracterizando doença hepática. Concluise baseado nos exames histopatológicos que as concentrações oferecidas de sementes de C. spectabilis 0,2, 0,4 e 0,6% no período de 28 dias são tóxicas aos cães / Abstract: It is known that C. spectabilis is widely used as green manure and for controlling nematodes, with that spreads easily, mingling with the infecting maize and other cereal rations. The aim of this study was to evaluate the toxic effects of Crotalaria spectabilis seeds, when crushed and mixed with the food of dogs. In this experiment we used 12 dogs, six males and six females, of various ages, which were divided into three groups of four animals. Each group received respectively 0.2, 0.4 and 0.6% of seeds of C. spectabilis, for 28 days, which were crushed and added to the diet. Weekly it was evaluated the hematological and biochemical profile of dogs, in addition to physical examination performed daily. In biochemical tests performed, significant alteration of the enzyme GGT. Clinical signs observed were mainly periocular alopecia and diffuse alopecia, and hyperkeratosis on nose, sneezing, coughing, occasional vomiting, constipation and dry feces. For the histopathology was extracted through a liver biopsy needle type TRU-CUT®, it was observed tumefactive hepatocytes, vacuolization of cells in the sinusoids, focal necrosis, anisocytosis cariomegalia, Kupffer cells with different pigmentation, fibrosis and presence of bile pigment in hepatocytes, characterizing liver disease. It was based on histopathological examinations offered that concentrations of seeds of C. spectabilis 0.2, 0.4 and 0.6% during the 28 days are toxic to dogs / Mestre

Orientador: Wilter Ricardo Russiano Vicente / Banca: Marcelo Emílio Beletti / Banca: José Octávio Jacomini / Banca: Maria Rita Pacheco / Banca: Paulo Henrique Franceschini / Resumo: O objetivo deste estudo foi avaliar histologicamente as biópsias testiculares de cães obtidas por punção aspirativa por agulhas fina (PAAF) e "tru cut". Foram utilizados 40 cães, adultos, hígidos, distribuídos em 2 grupos: G1 - punção biópsia aspirativa; e G2 - biópsia com agulha "tru-cut". Cada grupo foi dividido em quatro subgrupos (Ga, Gb, Gc e Gd) com cinco animais cada, sendo estes orquiectomizados 3, 7, 14 e 62 dias após as biópsias PAAF ou "Tru-cut". O material colhido pela PAAF foi submetido à avaliação citológica, e o proveniente da biópsia "tru-cut" e orquiectomia submetidos à histopatologia. Foram avaliados os espermiogramas e mensuração de comprimento e largura do escroto e colhido o soro para pesquisa de anticorpo antiespermatozóides. Referente ao espermiograma e a mensuração do escroto não foram observados diferenças (p>0,05) significativas. A amostra direcionada para citologia e histologia obtida pelas duas técnicas foi considerada de quantidade suficiente para diagnóstico. Na histopatologia a PAAF apresentou menor área de lesão e reação inflamatória comparada a "tru cut", no entanto esta última apresentou maior quantidade de material, preservando a arquitetura dos túbulos seminíferos e interstício. Não foi observado diferença (p>0,05) na produção de anticorpos anti-espermatozóides, após as biopsias. Concluímos que apesar das biópsias fornecerem material em qualidade e quantidade adequadas, e da PAAF ter se mostrado menos traumática, a escolha da técnica a ser empregada dependerá da finalidade para o qual o material se destina. / Abstract: The aim of this study it was evaluate hystologicaly the testicular biopsies in dogs obtained by aspirative puncture by fine needle and "tru-cut". Forty males dogs, adults, healthies were used, distributed in 2 groups: G1 - Fine Needle aspiration; and G2- biopsy by "Tru-cut". Each group were shared in four groups (Ga, Gb, Gc e Gd) with 5 animals each, and all of then were orchiectomized after 3, 7, 14, 62 days after the biopsies by FNA and "tru-cut" being performed. The samples collected by FNA were submitted to cytology evaluate, and the sample from tru-cut biopsy and orchiectomy submitted to histopathology. The spermiograms were evaluated, the testicular length and with were measure, and the animal's blood were collected to the anti-sperm antibody quantification. About the spermogram and measurement of scrotal bag any significative difference was observed (p>0,05). The sample to cytology and histology obtained from two techniques showed be in quantity enough to diagnostic. In histopathology, the biopsy FNA showed smaller damage area and inflammatory reaction compared to tru-cut, however this one showed biggest quantity of material, preserving the seminiferous tubules architeture and interstitium. It was not observed significative difference at anti-sperm antibodies production. We concluded that althought the biopsies provide material in quantity and quality appropriate, and the PAAF showed less traumatic, the choose of the technique to be used will depend the purpose for which the material is intended. / Doutor

<p>In the process of designing an electric power supply system for an aircraft, parameters like low weight and low losses are important. Reliability, robustness and low cost are other important factors. In the Saab Gripen aircraft, the design of the primary power supply of the electric flight control system was updated by exchanging a switching transistor regulator to a magnetic amplifier (magamp). By introducing a magamp design, weight was saved and a more reliable power supply system at a lower cost was achieved.</p><p> In this particular case, with the power supply of the electric flight control system in the Saab Gripen fighter, advantage could be taken of a specific permanent magnet generator (PM-generator). The frequency of the generator offered the perfect conditions for a magamp controller. A key parameter in designing magnetic amplifiers (magamps) is low losses. New amorphous alloys offer new possibilities of the technique in designing magnetic amplifiers, because of their extremely low losses.</p><p> The core losses are evaluated by studying the equations and diagrams specifying the power losses. The core losses are evaluated and compared with the copper losses in the process of optimizing low weight and low losses. For this an engineering tool is developed and demonstrated.</p><p> Evaluations of the hysteresis characteristics for the magnetic alloys, as well as modeling and simulation of the core losses, are presented in this work. The modeling of the core losses includes hysteresis losses, eddy current losses and excess losses as well as copper losses. The losses are studied dynamically during realistic operational conditions. The model can be used for any generic analysis of hysteresis in magnetic circuits. Applications of magnetic amplifiers in aircrafts have been demonstrated to be a feasible alternative</p>

In the process of designing an electric power supply system for an aircraft, parameters like low weight and low losses are important. Reliability, robustness and low cost are other important factors. In the Saab Gripen aircraft, the design of the primary power supply of the electric flight control system was updated by exchanging a switching transistor regulator to a magnetic amplifier (magamp). By introducing a magamp design, weight was saved and a more reliable power supply system at a lower cost was achieved. In this particular case, with the power supply of the electric flight control system in the Saab Gripen fighter, advantage could be taken of a specific permanent magnet generator (PM-generator). The frequency of the generator offered the perfect conditions for a magamp controller. A key parameter in designing magnetic amplifiers (magamps) is low losses. New amorphous alloys offer new possibilities of the technique in designing magnetic amplifiers, because of their extremely low losses. The core losses are evaluated by studying the equations and diagrams specifying the power losses. The core losses are evaluated and compared with the copper losses in the process of optimizing low weight and low losses. For this an engineering tool is developed and demonstrated. Evaluations of the hysteresis characteristics for the magnetic alloys, as well as modeling and simulation of the core losses, are presented in this work. The modeling of the core losses includes hysteresis losses, eddy current losses and excess losses as well as copper losses. The losses are studied dynamically during realistic operational conditions. The model can be used for any generic analysis of hysteresis in magnetic circuits. Applications of magnetic amplifiers in aircrafts have been demonstrated to be a feasible alternative / QC 20101103

Reverse genetics is an innovative molecular biology tool that enables the manipulation of viral genomes at the cDNA level in order to generate particular mutants or artificial viruses. The reverse genetics system for the influenza virus is arguably one of the best illustrations of the potential power of this technology. This reverse genetics system is the basis for the ability to regularly adapt influenza vaccines strains. Today, reverse genetic systems have been developed for many animal RNA viruses. Selection-free reverse genetics systems have been developed for the members of the Reoviridae family including, African horsesickness virus, bluetongue virus and orthoreovirus. This ground-breaking technology has led to the generation of valuable evidence regarding the replication and pathogenesis of these viruses. Unfortunately, extrapolating either the plasmid-based or transcript-based reverse genetics systems to rotavirus has not yet been successful. The development of a selection-free rotavirus reverse genetics system will enable the systematic investigation of poorly understood aspects of the rotavirus replication cycle and aid the development of more effective vaccines, amongst other research avenues. This study investigated the importance of co-expressed rotavirus proteins in the development of a selection-free rotavirus reverse genetics system. The consensus sequences of the rotavirus strains Wa (RVA/Human-tc/USA/WaCS/1974/G1P[8]) and SA11 (RVA/Simian-tc/ZAF/SA11/1958/G3P[2]) where used to design rotavirus expression plasmids. The consensus nucleotide sequence of a human rotavirus Wa strain was determined by sequence-independent cDNA synthesis and amplification combined with next-generation 454® pyrosequencing. A total of 4 novel nucleotide changes, which also resulted in amino acid changes, were detected in genome segment 7 (NSP3), genome segment 9 (VP7) and genome segment 10 (NSP4). In silico analysis indicated that none of the detected nucleotide changes, and consequent amino acid variations, had any significant effect on viral structure. Evolutionary analysis indicated that the sequenced rotavirus WaCS was closely related to the ParWa and VirWa variants, which were derived from the original 1974 Wa isolate. Despite serial passaging in animals, as well as cell cultures, the Wa genome seems to be stable. Considering that the current reference sequence for the Wa strain is a composite sequence of various Wa variants, the rotavirus WaCS may be a more appropriate reference sequence. The rotavirus Wa and SA11 strains were selected for plasmid-based expression of rotavirus proteins, under control of a T7 promoter sequence, due to the fact that they propagate well in MA104 cells and the availability of their consensus sequences. The T7 RNA polymerase was provided by a recombinant fowlpox virus. After extensive transfection optimisation on a variety of mammalian cell lines, MA104 cells proved to be the best suited for the expression rotavirus proteins from plasmids. The expression of rotavirus Wa and SA11 VP1, VP6, NSP2 and NSP5 could be confirmed with immunostaining in MA104 and HEK 293H cells. Another approach involved the codon-optimised expression of the rotavirus replication complex scaffold in MA104 cells under the control of a CMV promoter sequence. This system was independent from the recombinant fowlpox virus. All three plasmid expression sets were designed to be used in combination with the transcript-based reverse genetics system in order to improve the odds of developing a successful rotavirus reverse genetics system. Rotavirus transcripts were generated using transcriptively active rotavirus SA11 double layered particles (DLPs). MA104 and HEK293H cells proved to be the best suited for the expression of rotavirus transcripts although expression of rotavirus VP6 could be demonstrated in all cell cultures examined (MA104, HEK 293H, BSR and COS-7) using immunostaining. In addition, the expression of transcript derived rotavirus VP1, NSP2 and NSP5 could be confirmed with immunofluorescence in MA104 and HEK 293H cells. This is the first report of rotavirus transcripts being translated in cultured cells. A peculiar cell death pattern was observed within 24 hours in response to transfection of rotavirus transcripts. This observed cell death, however does not seem to be related to normal viral cytopathic effect as no viable rotavirus could be recovered. In an effort to combine the transcript- and plasmid systems, a dual transfection strategy was followed where plasmids encoding rotavirus proteins were transfected first followed, 12 hours later, by the transfection of rotavirus SA11 transcripts. The codon- optimised plasmid system was designed as it was postulated that expression of the DLP-complex (VP1, VP2, VP3 and VP6), the rotavirus replication complex would form and assist with replication and/or packaging. Transfecting codon- optimized plasmids first noticeably delayed the mass cell death observed when transfecting rotavirus transcripts on their own. None of the examined coexpression systems were able to produce a viable rotavirus. Finally, the innate immune responses elicited by rotavirus transcripts and plasmid-derived rotavirus Wa and SA11 proteins were investigated. Quantitative RT-PCR (qRT-PCR) experiments indicated that rotavirus transcripts induced high levels of the expression of the cytokines IFN- α1, IFN-1β, IFN-λ1 and CXCL10. The expression of certain viral proteins from plasmids (VP3, VP7 and NSP5/6) was more likely to stimulate specific interferon responses, while other viral proteins (VP1, VP2, VP4 and NSP1) seem to be able to actively suppress the expression of certain cytokines. In the light of these suppression results, specific rotavirus proteins were expressed from transfected plasmids to investigate their potential in supressing the interferon responses provoked by rotavirus transcripts. qRT-PCR results indicated that cells transfected with the plasmids encoding NSP1, NSP2 or a combination of NSP2 and NSP5 significantly reduced the expression of specific cytokines induced by rotavirus transcripts. These findings point to other possible viral innate suppression mechanisms in addition to the degradation of interferon regulatory factors by NSP1. The suppression of the strong innate immune response elicited by rotavirus transcripts might well prove to be vital in the quest to better understand the replication cycle of this virus and eventually lead to the development of a selection-free reverse genetics system for rotavirus. / PhD (Biochemistry), North-West University, Potchefstroom Campus, 2014

Reverse genetics is an innovative molecular biology tool that enables the manipulation of viral genomes at the cDNA level in order to generate particular mutants or artificial viruses. The reverse genetics system for the influenza virus is arguably one of the best illustrations of the potential power of this technology. This reverse genetics system is the basis for the ability to regularly adapt influenza vaccines strains. Today, reverse genetic systems have been developed for many animal RNA viruses. Selection-free reverse genetics systems have been developed for the members of the Reoviridae family including, African horsesickness virus, bluetongue virus and orthoreovirus. This ground-breaking technology has led to the generation of valuable evidence regarding the replication and pathogenesis of these viruses. Unfortunately, extrapolating either the plasmid-based or transcript-based reverse genetics systems to rotavirus has not yet been successful. The development of a selection-free rotavirus reverse genetics system will enable the systematic investigation of poorly understood aspects of the rotavirus replication cycle and aid the development of more effective vaccines, amongst other research avenues. This study investigated the importance of co-expressed rotavirus proteins in the development of a selection-free rotavirus reverse genetics system. The consensus sequences of the rotavirus strains Wa (RVA/Human-tc/USA/WaCS/1974/G1P[8]) and SA11 (RVA/Simian-tc/ZAF/SA11/1958/G3P[2]) where used to design rotavirus expression plasmids. The consensus nucleotide sequence of a human rotavirus Wa strain was determined by sequence-independent cDNA synthesis and amplification combined with next-generation 454® pyrosequencing. A total of 4 novel nucleotide changes, which also resulted in amino acid changes, were detected in genome segment 7 (NSP3), genome segment 9 (VP7) and genome segment 10 (NSP4). In silico analysis indicated that none of the detected nucleotide changes, and consequent amino acid variations, had any significant effect on viral structure. Evolutionary analysis indicated that the sequenced rotavirus WaCS was closely related to the ParWa and VirWa variants, which were derived from the original 1974 Wa isolate. Despite serial passaging in animals, as well as cell cultures, the Wa genome seems to be stable. Considering that the current reference sequence for the Wa strain is a composite sequence of various Wa variants, the rotavirus WaCS may be a more appropriate reference sequence. The rotavirus Wa and SA11 strains were selected for plasmid-based expression of rotavirus proteins, under control of a T7 promoter sequence, due to the fact that they propagate well in MA104 cells and the availability of their consensus sequences. The T7 RNA polymerase was provided by a recombinant fowlpox virus. After extensive transfection optimisation on a variety of mammalian cell lines, MA104 cells proved to be the best suited for the expression rotavirus proteins from plasmids. The expression of rotavirus Wa and SA11 VP1, VP6, NSP2 and NSP5 could be confirmed with immunostaining in MA104 and HEK 293H cells. Another approach involved the codon-optimised expression of the rotavirus replication complex scaffold in MA104 cells under the control of a CMV promoter sequence. This system was independent from the recombinant fowlpox virus. All three plasmid expression sets were designed to be used in combination with the transcript-based reverse genetics system in order to improve the odds of developing a successful rotavirus reverse genetics system. Rotavirus transcripts were generated using transcriptively active rotavirus SA11 double layered particles (DLPs). MA104 and HEK293H cells proved to be the best suited for the expression of rotavirus transcripts although expression of rotavirus VP6 could be demonstrated in all cell cultures examined (MA104, HEK 293H, BSR and COS-7) using immunostaining. In addition, the expression of transcript derived rotavirus VP1, NSP2 and NSP5 could be confirmed with immunofluorescence in MA104 and HEK 293H cells. This is the first report of rotavirus transcripts being translated in cultured cells. A peculiar cell death pattern was observed within 24 hours in response to transfection of rotavirus transcripts. This observed cell death, however does not seem to be related to normal viral cytopathic effect as no viable rotavirus could be recovered. In an effort to combine the transcript- and plasmid systems, a dual transfection strategy was followed where plasmids encoding rotavirus proteins were transfected first followed, 12 hours later, by the transfection of rotavirus SA11 transcripts. The codon- optimised plasmid system was designed as it was postulated that expression of the DLP-complex (VP1, VP2, VP3 and VP6), the rotavirus replication complex would form and assist with replication and/or packaging. Transfecting codon- optimized plasmids first noticeably delayed the mass cell death observed when transfecting rotavirus transcripts on their own. None of the examined coexpression systems were able to produce a viable rotavirus. Finally, the innate immune responses elicited by rotavirus transcripts and plasmid-derived rotavirus Wa and SA11 proteins were investigated. Quantitative RT-PCR (qRT-PCR) experiments indicated that rotavirus transcripts induced high levels of the expression of the cytokines IFN- α1, IFN-1β, IFN-λ1 and CXCL10. The expression of certain viral proteins from plasmids (VP3, VP7 and NSP5/6) was more likely to stimulate specific interferon responses, while other viral proteins (VP1, VP2, VP4 and NSP1) seem to be able to actively suppress the expression of certain cytokines. In the light of these suppression results, specific rotavirus proteins were expressed from transfected plasmids to investigate their potential in supressing the interferon responses provoked by rotavirus transcripts. qRT-PCR results indicated that cells transfected with the plasmids encoding NSP1, NSP2 or a combination of NSP2 and NSP5 significantly reduced the expression of specific cytokines induced by rotavirus transcripts. These findings point to other possible viral innate suppression mechanisms in addition to the degradation of interferon regulatory factors by NSP1. The suppression of the strong innate immune response elicited by rotavirus transcripts might well prove to be vital in the quest to better understand the replication cycle of this virus and eventually lead to the development of a selection-free reverse genetics system for rotavirus. / PhD (Biochemistry), North-West University, Potchefstroom Campus, 2014