Thesis (Ph. D.)--Ohio State University, 2005. / Title from first page of PDF file. Document formatted into pages; contains xv, 151 p.; also includes graphics. Includes bibliographical references (p. 134-151). Available online via OhioLINK's ETD Center

Thesis (M.S.)--University of Missouri-Columbia, 2004. / Typescript. Vita. Includes bibliographical references (leaves 63-65). Also available on the Internet.

Submitted by LIVEA MARIA GOMES (livea_pitti@hotmail.com) on 2016-02-02T12:14:18Z No. of bitstreams: 1 Dissertação Lívea M Gomes final com ficha (3).pdf: 1791851 bytes, checksum: eeb7f3ff7c0cffdcdb1eeb86397ed634 (MD5) / Approved for entry into archive by Sandra Manzano de Almeida (smanzano@marilia.unesp.br) on 2016-02-02T13:24:27Z (GMT) No. of bitstreams: 1 gomes_lm_me_bot.pdf: 1791851 bytes, checksum: eeb7f3ff7c0cffdcdb1eeb86397ed634 (MD5) / Made available in DSpace on 2016-02-02T13:24:27Z (GMT). No. of bitstreams: 1 gomes_lm_me_bot.pdf: 1791851 bytes, checksum: eeb7f3ff7c0cffdcdb1eeb86397ed634 (MD5) Previous issue date: 2015-12-11 / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / O objetivo do presente estudo foi avaliar os efeitos do aumento do nível de triptofano na dieta de leitões desmamados submetidos a diferentes programas de luz. Foram utilizados 72 leitões desmamados aos 21 dias de idade (peso inicial de 6,6+ 2,33 kg). O delineamento experimental foi o de blocos casualisados com 24 dias de duração em esquema fatorial 2 x 2 (PL: 12 ou 23h de luz/dia e dois NT digestível: 2,6 ou 5,2 g de L-Trp / kg de ração pré-inicial 1 (0-14 dias) e de 2,4 ou 4,8 g de L-Trp / kg na dieta pré-inicial 2 (15-24 dias), com 6 repetições e 3 animais por parcela. Foram avaliados o desempenho, glicemia, perfil plasmático do cortisol e comportamento dos animais. A glicose no sangue foi determinada nos dias 0 e 24, e o cortisol nos dias 0 e 8. O comportamento foi observado nos dias 2, 9, 16 e 23. Não houve interação entre PL e NT sobre desempenho e glicose. Nos períodos de 0-14 e 0-24 dias, os tratamentos não influenciaram o ganho diário de peso, consumo diário de ração e glicose no sangue, enquanto que a conversão alimentar melhorou (P<0,05) apenas no período de 0-14 dias para os animais que receberam 12 horas de luz/dia. Os tratamentos não influenciaram a frequência do comportamento “em movimento”, entretanto houve interação entre PL e NT para o cortisol plasmático, frequência dos comportamentos “ócio” e “alimentar”. Leitões que receberam o maior NT apresentaram menor nível de cortisol plasmático (P<0,05), menor comportamento “alimentar” (P<0,05) e maior comportamento “ócio” (P<0,05) em relação aos animais que receberam NT normal quando submetidos ao PL de 23 horas de luz/dia. O uso de PL de 23h de luz/dia não é indicado, entretanto, quando este manejo for adotado o maior nível de triptofano na dieta é recomendado. / The objective of the present study was to avaluate the effects of light programs (LP) and tryptophan levels (TL) in the diet on performance, blood glucose, plasma profile of cortisol and behavior. A total of 72 piglets weaned at 21 days of age (starting weight 2.33 kg 6,6±) was used. The experimental design was randomized blocks with 24 days duration in a factorial 2 x 2 (LP 12 or 23h light / day and two digestible TL: 2.6 or 5.2 g of L-Trp / kg diet on pre-starter 1 (0-14 days) and 2.4 or 4.8 g of L-Trp / kg diet on prestarter 2 (15-24 days), with 6 repetitions and 3 animals per pen. Glucose blood was determined on d 0 and 24, and cortisol on d 0 and 8. The behavior was observed on d 2, 9, 16 and 23. No effects of PL and NT interaction were observed on performance and blood glucose. In periods of 0- 14 and 0-24 days, the treatments did not influence the daily weight gain, daily feed intake and blood glucose, while feed conversion improved (P <0.05) only in the period of 0-14 days for piglets receiving PL 12h light / day. Treatments did not influence the frequency of “moving” behavior, however there was interaction between PL and NT for plasma cortisol, frequency of “feeding” and “lying” behavior. Piglets that received high NT had lower plasma cortisol level (P <0.05), lower “feeding” behavior (P <0 .05) and increased “lying” behavior (P <0.05) compared to animals that received normal NT when subjected to PL 23 hours light / day. The PL 23h light / day isn’t indicated for piglets, but when this happen, highest tryptophan level in the diet is recommended.

No description available.

The aim of this study was to characterize strains used in the cross-protection scheme in South Africa by establishing Polymerase Chain Reaction (PCR) systems aimed at differentiating the strains by targeting the conserved p23 gene and the variable 5' half of the Citrus tristeza virus (CTV) genome. Two cross-protecting sources GFMS 12 and GFMS 35; and eight single aphid sub-isolates were tested and classified into strain types or genotypes. An oligonucleotide microarray system was developed to differentiate T30 and T36 strains of CTV. The establishment and development of such tests will enable the South African Citrus Industry to better select mild strains for cross-protection and determine which strains are present in citrus growing areas so as to better understand the dynamics of the disease. The first aim was to characterise the p23 gene of possible mild-strain cross-protection isolates in South Africa (RSA) and compare them to known isolates worldwide. Isolates were amplified with bi-directional RT-PCR, sequenced and phylogenetic analysis performed. The predicted amino acid sequences were compared for areas of possible variability for further strain differentiation. A bi-directional PCR developed by Sambade et al. (2003) was established that targets differences in amino acid positions 78-80 of the p23 gene and allows discrimination of isolates into mild, atypical and severe groups. The group designations of RSA isolates 390-3 and 390-5 were atypical; 390-4, 389-4 and 389-3 were mild; GFMS 35 had mild and atypical isolates; GFMS12, 12-7 and 12-9 had mild and severe isolates and; 12-5 was severe. The three main clusters on the phylogenetic tree confirmed the group designations of these isolates. Isolates in the atypical group were more diverse than ones in the mild or severe groups. There were 53 polymorphic sites within the amino acid sequences of p23 gene of the RSA and reference isolates, of which 4 distinct regions showed variability. The amino acid region 78-80 was confirmed as being very useful in grouping these isolates as mild, severe or atypical. The PCR system was robust, reproducible and has potential in the RSA Citrus industry as a screening tool in selecting mild strains for cross-protection and in detecting mixed strains in isolates. The secondary aim was to establish the 23 primer pair PCR system developed by Hilf et al. (2000) to differentiate isolates as T36, T30, VT or T3 genotypes. Each isolate was tested with RT-PCR using 23 individually optimised genotype specific primer sets (Hilf et al., 2000). The most common genotype detected was T30 and the least common was T3. The GFMS 35, T30 plant and 389-3 isolates had a homogenous T30 genotype profile and isolate 12-5 had a VT genotype profile. The 389-4, 390-3 and 390-4 isolates had a predominantly T30 genotype profile and isolates 12-7 had a predominantly VT genotype profile. Isolate GFMS 12 had a mixed genotype profile indicative of a mixed infection while isolates 390-5 and 12-9 appeared to have mixed genotypes of VT, T30 and T36. Isolates 390-3 390-4 and 390-5 had no amplification within regions 4-7 and appear to be highly variable isolates or possible recombinants. The T3 genotype specific markers were found in region 2 of a few isolates and could be a cross-reacting primer set to the T3o genotype. It is useful for homogenous strains in determining the genotypes, molecular marker information, possible variability or recombination and for approving isolates for mild strain cross-protection. Potential drawbacks of the system include non-amplification of regions; cross-reacting primers; difficulty in optimising; and secondary structures. It was difficult to objectively draw conclusions if an isolated had mixed genotypes since mixed genotype amplifications were not consistently found in all regions targeted. The third aim was to develop an oligonucleotide (oligo) microarray system to differentiate mild T30 and severe T36 strains. The 5' half of the CTV genome was Cy3 5'-end labelled and amplified. Oligos were designed to be T36-strain specific with a Tm above 60 °C and if possible a GC content above 65 %; and differed in amount and position of mismatches to strain T30. A standard operating protocol was set up by testing different labelling methods, hybridization mixes and washing steps. The array was tested using individual T30 and T36 strains as templates at 42, 52 and 60 °C. Experimental variation was quantified and normalised. The secondary structures of the hybridizing amplicons were determined by mfold (Zuker et al., 2003). Some oligos were specific at 42 °C and others at 52 °C. The hybridization allowed a clear differentiation of strain T36 with 13 of the T36-specific oligos at their optimal hybridization temperature. A few oligonucleotides showed cross-hybridization to strain T30 and were not used in further analysis. Oligonucleotides with 21 % or more mismatches were successful oligos, whereas ones that had 18 % or fewer mismatches had cross-hybridization. Some oligos were modified to include Locked Nucleic Acid (LNA) instead of DNA in an attempt to increase specificity with two of them having increased specificity compared to the unmodified DNA oligonucleotides. The successful differentiation by hybridization to strain specific oligos opens paths for highly parallel, yet specific assays for strain differentiation of CTV strains and a more thorough insight into the future strains circulating in RSA. / Dissertation (MSc (Microbiology))--University of Pretoria, 2008. / Microbiology and Plant Pathology / unrestricted

It is fairly well known that alpha-amino dicarboxylic acids combine quite readily with basic metal ions such as the alkaline earths. The nature of the problem includes the synthesis and characterization of six platinum metal chelates of two alpha-animo dicarboxylic acids, namely aspartic and glutamic acids. The metal ion upon which the investigation is focused are platinum (II), palladium (II), and rhodium (III). Several of these are reported as having been repaired. The series is incomplete, however, and little study has been done correlating the abilities and trends of the complexes as a function of the metal ion and carbon chain length of the ligand acid molecule. Hence, the ultimate aim of the investigation is to study the tendency of chelation as the carbon chain length of the alpha-amino dicarboxylic acid increases and also to study the stability of the chelates as the central metal ion is varied. Because there are numerous problems involved here, the investigation extends somewhat beyond the scope of a Master’s research. Therefore the investigation is limited only to aspartic and glutamic acid. The methods of synthesis for the palladium, platinum, and rhodium chelates of aspartic and glutamic acids were studied and outlined. After synthesis, the means which were employed for characterization included elemental analysis, molecular weight determination, and infrared and ultraviolet spectra.

Thermus bacteria are of special interest because of their ability to live in high temperature environments. Their enzymes exhibit higher and stable activity in industry as compared to mesophilic or synthetic counterparts. Thermus bacteria are capable of reducing heavy metals which is essential in eradication of heavy metal pollution and controlling global warming. Genome rearrangements were investigated in Thermus species and the extent to which they affected the distribution of functionally related genes on the chromosome and its implication on the coherence of the metabolic network. The contribution of horizontal gene transfer to genome rearrangements and the shuffling of genes on the chromosome were analysed. Horizontally transferred genes were identified alongside their donors and recipients, their age and relative time of insertion. Metabolic networks were clustered and compared to determine the extent to which they were affected by rearrangements as a measure of evolutionary pressures experienced by organisms. Factors that enhance protein thermostability were analysed by determining dominant substitutions for amino acid residues and their properties. Protein thermostability was measured using the UNAFold algorithm. Amino acid substitutions were compared between less and highly thermophilic orthologous sequences in T. scotoductus SA-01 and T. thermophilus HB27 respectively. Protein structures were modelled for orthologs that met a defined selection criterion. Dominant amino acid substitutions were analysed in the structures to determine their locations. The contribution of dominant substitutions to energy changes in structures was analysed using FoldX program. Results revealed a uniform distribution of functionally related genes among thermophilic and mesophilic organisms. The contribution of horizontal gene transfer to genome rearrangements was found to be insignificant. Metabolic networks for Thermus species were poorly clustered in correlation with their optimum environmental growth temperatures. Non-polar, small and charged amino acids were found to significantly enhance thermostability. Higher occurrence of alanine substituted by serine and threonine; and arginine substituted by glutamine and lysine were observed to influence thermostability. Structural comparison showed that mutations were mostly located on the surfaces and helices of structures. The positions of mutations appeared to influence their energy contribution to the overall structure as measured by FoldX algorithm. / Thesis (PhD)--University of Pretoria, 2013. / gm2014 / Biochemistry / unrestricted

All twenty amino acids did not appear simultaneously in nature. Instead some of them appeared early, while others were added into the genetic code later. The amino acids that were formed by Miller (1953) are suggested to have appeared early in evolutionary history, and the amino acids associated with codon capture developed late in the course of evolution. The chronological order of appearance of the amino acids proposed by Trifonov (2000) was G/A, V/D, P, S, E/L, T, R, N, K, Q, I, C, H, F, M, Y, W. According to Romero et al. (1997) amino acids G, D, E, P and S are disorder-promoting residues and C, F, W and Y are order-promoting residues this means that the early or the ancient amino acids were disorder promoting and the order promoting residues came late into the genetic code. These observations led to the hypothesis that the first proteins, which were comprised of the early amino acids only, were disordered, and, furthermore, that the appearance of the late amino acids and the appearance of the structural proteins were concurrent. Software developed by Brooks et al. (2004) to find the amino acid composition of the LUA (Last Universal Ancestor) was used to test this hypothesis. For this work, the Clusters of Orhtologous Groups of proteins (65 COGs) were split into enzymes and non-enzymes. It was found that intrinsic disorder was abundant in both the groups of proteins, with non enzymes being much more disorder than enzymes. Further analysis was done to check for the frequency of the modern amino acids C, F, W, and Y in the Protein data bank (PDB) and Swissprot.

L- Histidine is a very important amino acid and is widely distributed in living systems. In the growth and multiplication of animal cells, it was found that L-histidine is one of the amino acids that must be present. From the kinetic studies of certain enzymes such as chymotrypsin and ribonuclease, it has been proposed that the imidazolyl group of the histidine residue may serve as the basic electron donor, and that the histidine residue in some cases may be the active site of the enzyme. The interactions of L-histidine with heavy metal ions, including Ni (II) (1), Zn (II) (1, 2), Fe (II) (1), Cu (II) (3, 4, 5, 7, 9, 10), Mn (II) (6), Cd (II) (3, 4) and Co (II) (1, 11, 12), have been investigated.

京都大学 / 0048 / 新制・課程博士 / 博士(工学) / 甲第21276号 / 工博第4504号 / 新制||工||1700(附属図書館) / 京都大学大学院工学研究科合成・生物化学専攻 / (主査)教授 跡見 晴幸, 教授 森 泰生, 教授 梅田 眞郷 / 学位規則第4条第1項該当 / Doctor of Philosophy (Engineering) / Kyoto University / DGAM