No description available.

In the context of the somatic mutation theory of chemical carcinogenesis, mutations are thought to arise during the replication of DNA past carcinogen-DNA adducts. The work described in this thesis deals with the testing of a hypothetical mechanism whereby mammalian cells are able to replicate their DNA past polycyclic aromatic hydrocarbon DNA adducts. The second objective of this thesis work was to develop both in vivo and in vitro models to study the induction of mutations in a target human gene by chemical carcinogens from two different classes, polycyclic aromatic hydrocarbons and nitrosamines. To approach the hypothetical mechanism of bypass replication in mammalian cells, synchronized Chinese hamster ovary cells were treated with the ultimate carcinogenic form of benzo (a) pyrene, 7β, 8α-dihydroxy-9α, 10α-epoxy-7,8,9,10-tetrahydrobenzo (a) pyrene (BPDE I). Using the pH step alkaline elution assay, it was found that the reduced rate of S phase progression was due to a delay in the appearance of multiple replicon size nascent DNA. It was determined using agarose gel electrophoresis that the ligation of Okazaki size replication intermediates was blocked in BPDE I-treated, synchronized CHO cells. The data obtained were, therefore, supportive of the 'block-gap' model of DNA bypass replication in carcinogen damaged mammalian cells. To study mutagenesis of a specific sequence induced by chemical carcinogens, the human c-Ha-ras proto-oncogene was transfected into the mouse fibroblast cell line, NIH 3T3. Transfected NIH 3T3 cell lines (HHRN 1-4) were isolated that had a low copy number of the human c-Ha-ras proto-oncogene and a non-transformed phenotype. It was determined that the integrated human c-Ha-ras gene was hypomethylated, and expressed at the messenger level. The human c-Ha-ras protein, p21, was also detected in these transfected cell lines. Treatment of the HHRN cell lines with the nitrosamine, N-methyl-N-nitroso-N'-nitroguanidine (MNNG) resulted in transformed NIH 3T3 foci. In vitro MNNG treatment of the plasmid, z-6, and transfection into NIH 3T3 cells led to the isolation of transformed cell lines. Screening of the in vitro and in vivo treated, transformed cell lines by RNA:RNA duplex mismatch analysis led to the detection of no mutations within the first exon of the human c-Ha-ras oncogene.

The overriding objective of this study was to induce genetic variation in Vernonia (Vernonia galamensis variety ethiopica) using ethylmethanesulfonate (EMS) and select mutants for subsequent selective breeding. Vernonia is an underutilised, potential novel oilseed crop with multiple applications in industry mostly due to the production of naturally epoxidised vernolic acid oil. Commercial cultivation of vernonia is significantly hampered by non-uniform seed maturity, tall plant height, seed shattering and lack of appropriate technologies for mechanical harvesting, seed threshing and cleaning. Mutations of a single or few genes possessing target traits are invaluable in crop improvement programs. Chemical mutagenesis using EMS is an important, affordable and effective method to induce random useful genetic mutations in crop plants. Ethylmethanesulfonate mutagenesis has previously been reported to affect various agronomic traits, induce a wide variety of phenotypic mutations and alter both seed oil content and fatty acid profile on several crops. The objectives of this study were: (i) to determine an optimum EMS treatment combination i.e. exposure duration, temperature and dose that would enable 50-60% germination at minimum days to emergence in selected V. galamensis var. ethiopica lines (Vge-1, Vge-4, Vge-7 and Vge-10), (ii) to induce genetic variation using predetermined optimal treatment conditions and select mutants in V. galamensis variety ethiopica lines (Vge-1 and Vge-4) and (iii) to evaluate oil content and fatty acid compositions among seeds of chloroplast mutants, EMS treated seeds and untreated controls of Vge-1 and Vge-4. Before any mutation is administered in plants, it is important that the optimal mutation dose is determined. The lethal dose 50 (LD50) was the standard used in this study to find optimal treatment conditions. Significant interactions (P<0.001) existed between EMS, line, time and temperature with respect to days to 50% emergence, germination percentage and seedling height. Optimal days to 50% emergence (10-12 days) and germination (50- 58%) was achieved for Vge-1, Vge-7 and Vge-10 when treated with 0.372% EMS at 350C for 1 hour treatment. The optimal treatment combination for Vge-4 was 0.372% EMS at 32.50C for 2hr. The treatment combinations that yielded optimum results in the tested lines were utilized to induce large scale mutations in V. galamensis to select target mutants in the field. Large scale mutation was conducted using the observed optimal treatment conditions. Ethylmethanesulfonate mutagenesis significantly delayed days to head formation, days to flowering and days to maturity on both lines. Delays in days to emergence were only significant in Vge-4. EMS treatment also significantly reduced germination percentage, number of seeds per head, number of fertile plants, plant height and plot yield for both Vge-1 and Vge-4. Thousand seed weight significantly increased in treated seeds of the two lines. Chlorophyll mutants were observed for tested lines associated with high count of sterility for both lines. Ethylmethanesulfonate successfully induced phenotypic mutation in selected vernonia lines, however at this stage the effect of mutation on vernonia seed oil content and fatty acid was unknown. Liquid gas chromatography method was employed for oil and fatty acids analysis. In Vge-1, significant differences were observed in composition of linoleic and oleic acid due to the mutagenesis. Significant increases in linoleic and oleic acid composition were found in chloroplast mutants due to EMS mutagenesis. No significant differences were detected in fatty acid compositions in Vge-4 after the EMS treatment. Differential responses were observed when lines were compared at various EMS mutation levels showing significant effect on vernolic, linoleic and oleic acids compositions. In both lines no differences were detected on seed oil content, palmitic acid, steraic acid and arachidic acid compositions after the treatment. Oil content significantly and positively correlated with vernolic acid for Vge-1 (P<0.001; r= 0.898) and Vge-4 (P<0.05; r= 0.65). Vernolic acid significantly and negatively correlated with other fatty acids. The study found that EMS mutagenesis significantly changed the oleic acid and linoleic acid compositions in vernonia. However, the oil content and vernolic acid composition were not significantly affected by EMS treatment. This study established that EMS was successful in inducing genetic variation (in agronomic traits, seed oil content and fatty acid composition) in the two tested lines of V. galamensis. Data from a single planting generation is insufficient to conclude fully on the effect of EMS on V. galamensis; therefore it is highly recommended that further multigenerational studies should be conducted with an increased number of testing lines from a wide range of environmental backgrounds. / Thesis (M.Sc.Agric.)-University of KwaZulu-Natal, Pietermaritzburg, 2012.

by Rebecca McGreal. / Vita. / Thesis (Ph.D.)--Florida Atlantic University, 2012. / Includes bibliography. / Electronic reproduction. Boca Raton, Fla., 2012. Mode of access: World Wide Web. / The central premise of this dissertation is that the small heat shock protein (sHSP), (Sa(BB-crystallin is essential for lens and retinal pigmented epithelial (RPE) cell function and oxidative stress defense. To date, the mechanism by which it confers protection is not known. We hypothesize that these functions could occur through its ability to protect mitochondrial function in lens and RPE cells. To test this hypothesis, we examined the expression of (Sa(BB-crystallin/sHSP in lens and RPE cells, we observed its localization in the cells, we examined translocation to the mitochondria in these cells upon oxidative stress treatment, we determined its ability to form complexes with and protect cytochrome c (cyt c) against damage, and we observed its ability to preserve mitochondrial function under oxidative stress conditions in lens and RPE cells. In addition to these studies, we examined the effect of mutations of (Sa(BB-crystallin/sHSP on its cellular localization and translocation patterns under oxidative stress, its in vivo and in vitro chaperone activity, and its ability to protect cyt c against oxidation. Our data demonstrated that (Sa(BB-crystallin/sHSP is expressed at high levels in the mitochondria of lens and RPE cells and specifically translocates to the mitochondria under oxidative stress conditions. We demonstrate that (Sa(BB-crystallin/sHSP complexes with cyt c and protects it against oxidative inactivation. Finally, we demonstrate that (Sa(BB-crystallin/sHSP directly protects mitochondria against oxidative inactivation in lens and RPE cells. Since oxidative stress is a key component of lens cataract formation and age-related macular degeneration (AMD), these data provide a new paradigm for understanding the etiology of these diseases.

Among the available methods of creating selectable variation, induced mutagenesis has been historically under-utilized in cotton improvement. Dick Auld showed that chemical mutagenesis could be used to enhance fiber length of a medium staple cotton cultivar without sacrificing yield. The goal of this project was to determine if mutagenesis could be used to improve the fiber quality of a germplasm line already considered to be at the upper-limits of fiber length. Seed of TAM 94 L-25 were treated with EMS in 2001 and the M2 generation was produced at Lubbock, Texas in 2002. More than 1200 M3 plants were grown at College Station, Texas in 2004, harvested individually, and HVI fiber properties determined. The top and bottom 1 percent for UHML, strength, and elongation were selected and the seeds of these individuals planted as an M4 progeny row nursery in 2005. Approximately ten individual plants per progeny row were harvested for re-evaluation of fiber parameters. From the approximately 1600 individual TAM 94L-25 M4 plants harvested in 2005, 61 were selected and subsequently treated as pure lines. Agronomic performance trials were conducted on 61 of those TAM 94L-25 M lines along with the M0 check and two commercial cultivar checks, Fiber Max 832 and Phytogen 355, in 2008 and 2009 in College Station and Weslaco, Texas. Within-boll yield components were examined for 13 representative mutant lines and the three checks. TAM 94L-25 averaged 751 kg lint ha^-1, 31.1 mm UHML, 303 kN m kg^-1 fiber bundle strength, and 6.0 percent elongation. The 61 mutant lines yielded from 366 to 932 kg lint ha^-1, exhibited UHML from 24.3 to 34.9 mm, fiber bundle strengths of 261 to 333 kN m kg^-1, and elongations from 5.4 to 8.1 percent. Seed surface area of the TAM 94L-25 M-lines was between 99 and 124 mm^2, and fibers per unit seed surface area from 123 to 168 fibers mm^-2. The M0 parent, TAM 94L-25 averaged 125 mm^2 seed-1, and 128 fibers mm^-2. The data presented herein demonstrate that EMS-induced mutagenesis was successful in creating TAM 94L-25 M-lines with superior fiber and yield traits to that of the non-mutated, high fiber quality parent, TAM 94L-25.

No description available.

Bibliography : leaves i-xxiii. / vi, 95, xxiii, [170] leaves, [4] leaves of plates : ill. (some col.) ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Thesis (Ph.D.)--University of Adelaide, Dept. of Clinical and Experimental Pharmacology, 1991

Thesis (Ph.D.)--McMaster University (Canada), 1996. / Source: Dissertation Abstracts International, Volume: 58-06, Section: B, page: 3006. Adviser: B. E. McCarry.

Thesis (Ph.D.) - University of KwaZulu-Natal, Pietermaritzburg, 2009. / Submitted to the African Centre for Crop Improvement. Full text also available online. Scroll down for electronic link.

Orientador: Chung Man Chin / Coorientador: Jean Leandro dos Santos / Banca: Gustavo Henrique Goulart Trossini / Banca: Priscila Longhin Bosquesi / Resumo: Doenças neurodegenerativas são definidas como distúrbios que acarretam na perda progressiva de estrutura e/ou função, e até mesmo morte de células neuronais especificas, em diferentes distribuições anatômicas, e, consequentemente, dando origem a diferentes sintomas clínicos. Entre as varias doenças neurodegenerativas a doença de Alzheimer (DA) é a principal e a mais comum forma de demência. A DA é uma condição progressiva complexa que envolve interações sequenciais de cascatas patológicas, incluindo a interação dos peptídeos β-amiloides (βA) que se agregam levando ao desenvolvimento de placas senis, e a hiperfosforilação e agregação da proteína tau formando emaranhados neurofibrilares (ENF), juntamente a processos associados, como a inflamação e o estresse oxidativo. O fator de necrose tumoral alfa (TNF-α) é uma das principais citocinas pró-inflamatórias cujos níveis elevados são observados em diversas doenças neurodegenerativas. O óxido nítrico (NO) é uma importante molécula de sinalização na modulação da plasticidade sináptica e está associada ao processo de aprendizagem e memória, assim como seus efeitos neuroprotetores quando presente em baixas concentrações. Assim, compostos derivados ftalimídicos, capazes de inibir TNF-α, poderiam reduzir o processo de neuroinflamação que é descrita na DA, retardando a progressão da doença, e derivados doadores de NO poderiam contribuir na plasticidade sináptica, cognição e memória dos pacientes acometidos pela DA. Foi objetivo deste trabalho a obtenção dos compostos Lapdesf FUR-FD I-VI, planejados por meio da estratégia de hibridação molecular entre a subunidade ftalimídica e o núcleo furoxânico. Os compostos obtidos foram avaliados para determinar o potencial efeito anti-inflamatório através do modelo de inflamação aguda (edema de pata), a determinação do Log P experimentalmente para avaliar sua capacidade de permear a barreira ... / Abstract: Neurodegenerative diseases are defined as disorders that result in progressive loss of structure and/or function, and even cell death in neuronal specific in different anatomical distributions, and hence giving rise to varying clinical symptoms. Among the various neurodegenerative diseases Alzheimer's disease (AD) is the main and most common form of dementia. AD is a progressive condition that involves complex interactions and sequential pathological cascades, including the interaction of β-amyloid peptide (βA) which clump together leading to the development of senile plaques, and hyperphosphorylation and aggregation of tau forming neurofibrillary tangles (ENF) along the associated processes, such as inflammation and oxidative stress. The tumor necrosis factor alpha (TNF-α) is a major proinflammatory cytokines whose high levels are observed in several neurodegenerative diseases. Nitric oxide (NO) is an important signaling molecule in the modulation of synaptic plasticity and its correlation with the process of learning and memory, as well as its neuroprotective effects when present in low concentrations. Thus phthalimidic derived compounds capable of inhibiting TNF-α , could reduce the neuroinflammation process that is described in AD, slowing the disease progression, and NO donors derived may contribute to synaptic plasticity, cognition and memory of patients affected by AD. This work aims to obtain the compounds (Lapdesf FUR-FD I-VI), designed by molecular hybridization strategy between the subunit and core phthalimidic and furoxan. The compounds obtained were evaluated for potential anti-inflammatory effect through the model of acute inflammation (paw edema), their ability to permeate the blood-brain barrier (BBB) using a tool of the partition coefficient (log P) obtained experimentally mutagenicity with the in vivo micronucleus test in reticulocytes obtained in peripheral blood, the ability of the compounds to inhibit the ... / Mestre