Orientador: Julio Cezar de Melo Castilho / Banca: Orivaldo Tavano / Banca: Luiz Cesar de Moraes / RESUMO: O objetivo nesta pesquisa foi estimar o crescimento e desenvolvimento esqueletal observando radiograficamente as alterações morfológicas da primeira, segunda e terceira vértebras cervicais , de acordo com o método proposto por Hassel & Farman32 (1995) em pacientes que se encontravam no surto de crescimento puberal. A inspeção radiográfica das vértebras cervicais foi realizada por meio de telerradiografias laterais e o surto de crescimento puberal foi identificado através dos eventos de ossificação da mão e punho. Para tanto, foram selecionadas as telerradiografias laterais e radiografias carpais de 110 brasileiros leucodermas de ambos os sexos, com idade cronológica variando dos 8 a 14,6 anos para as meninas e dos 9,5 aos 15,4 anos para os meninos. Os resultados revelaram que houve correlação estatisticamente significante entre os indicadores de maturação das vértebras cervicais (IMVC) e aqueles pacientes que se encontravam no surto de crescimento puberal. Concluiu-se que a avaliação radiográfica das alterações morfológicas das vértebras cervicais, nas telerradiografias laterais, constitui um parâmetro alternativo, confiável e prático na avaliação esquelética, vindo a complementar a gama de informações que se deve obter do paciente em tratamento ortodôntico e, circunstancialmente, substituir outros métodos de avaliação / ABSTRACT: The aim of this research was to estimate the skeletal growth and development by observingradiographically the morphological changes of the first, second and third cervical vertebrae, according to the method proposed by Hassel &Farman32 (1995) in patients who were going through the outbreak of pubertal growth. The radiographic inspection of the cervical vertebrae was performed by means of cephalometric radiographs and the outbreak of pubertal growth was identified through the events of ossification of the hand and wrist. For that purpose, we selected thecephalometric radiographs and hand•wrist radiographs of 110 Brazilianleukoderms, male and female,with chronological age ranging from 8 to 14,6 years old for the girls and from 9,5 to 15,4 years old for the boys.The results showed that there was a statistically significant correlation between the maturation indicators of the cervical vertebrae and the patients who were going through the outbreak of pubertal growth. The conclusion was that the radiographic evalution of the changes of the cervical vertebrae, in the cephalometric radiographs, is an alternative parameter, which is reliable and practical in the skeletal evalution, and the it completes the range of information that must be obtained from the patient under orthodontic treatment and that it can, circumstantially, replace other methods of evalution / Mestre

Thesis (doctoral)--Universität Zürich, 1956.

Thesis (doctoral)--Universität Zürich, 1956.

Made available in DSpace on 2014-11-10T11:10:01Z (GMT). No. of bitstreams: 0 Previous issue date: 2000-03-31Bitstream added on 2014-11-10T11:57:19Z : No. of bitstreams: 1 000138226.pdf: 2159909 bytes, checksum: 304da4f97692e3030d7ee688e8eb17ad (MD5) / O objetivo nesta pesquisa foi estimar o crescimento e desenvolvimento esqueletal observando radiograficamente as alterações morfológicas da primeira, segunda e terceira vértebras cervicais , de acordo com o método proposto por Hassel & Farman32 (1995) em pacientes que se encontravam no surto de crescimento puberal. A inspeção radiográfica das vértebras cervicais foi realizada por meio de telerradiografias laterais e o surto de crescimento puberal foi identificado através dos eventos de ossificação da mão e punho. Para tanto, foram selecionadas as telerradiografias laterais e radiografias carpais de 110 brasileiros leucodermas de ambos os sexos, com idade cronológica variando dos 8 a 14,6 anos para as meninas e dos 9,5 aos 15,4 anos para os meninos. Os resultados revelaram que houve correlação estatisticamente significante entre os indicadores de maturação das vértebras cervicais (IMVC) e aqueles pacientes que se encontravam no surto de crescimento puberal. Concluiu-se que a avaliação radiográfica das alterações morfológicas das vértebras cervicais, nas telerradiografias laterais, constitui um parâmetro alternativo, confiável e prático na avaliação esquelética, vindo a complementar a gama de informações que se deve obter do paciente em tratamento ortodôntico e, circunstancialmente, substituir outros métodos de avaliação / The aim of this research was to estimate the skeletal growth and development by observingradiographically the morphological changes of the first, second and third cervical vertebrae, according to the method proposed by Hassel &Farman32 (1995) in patients who were going through the outbreak of pubertal growth. The radiographic inspection of the cervical vertebrae was performed by means of cephalometric radiographs and the outbreak of pubertal growth was identified through the events of ossification of the hand and wrist. For that purpose, we selected thecephalometric radiographs and hand•wrist radiographs of 110 Brazilianleukoderms, male and female,with chronological age ranging from 8 to 14,6 years old for the girls and from 9,5 to 15,4 years old for the boys.The results showed that there was a statistically significant correlation between the maturation indicators of the cervical vertebrae and the patients who were going through the outbreak of pubertal growth. The conclusion was that the radiographic evalution of the changes of the cervical vertebrae, in the cephalometric radiographs, is an alternative parameter, which is reliable and practical in the skeletal evalution, and the it completes the range of information that must be obtained from the patient under orthodontic treatment and that it can, circumstantially, replace other methods of evalution

Over the past couple of years there has been rapid progress in the development and design of safe and effective delivery systems for the administration of protein and peptide drugs. The effective delivery of these type of drugs are not always as simple as one may think, due to various inherent characteristics of these compounds. Due to the hydrophilic nature and molecular size of peptide and protein drugs, such as recombinant human growth hormone, they are poorly absorbed across mucosal epithelia, both transcellularly and paracellularly. This problem can be overcome by the inclusion of absorption enhancers in peptide and protein drug formulations but this is not necessarily the best method to follow. This investigation focussed specifically on the evaluation of the ability of the PheroidTM carrier system to transport recombinant human growth hormone across mucosal epithelia especially when administered via the nasal cavity. The PheroidTM delivery system is a patented system consisting of a unique submicron emulsion type formulation. The PheroidTM delivery system, based on PheroidTM technology, will for ease of reading be called Pheroid(s) only throughout the rest of this dissertation. The Pheroid carrier system is a unique microcolloidal drug delivery system. A Pheroid is a stable structure within a novel therapeutic system which can be manipulated in terms of morphology, structure, size and function. Pheroids consist mainly of plant and essential fatty acids and can entrap, transport and deliver pharmacologically active compounds and other useful substances to the desired site of action. The specific objectives of this study can be summarised as follows: a literature study on Pheroid technology; a literature study on chitosan and N-trimethyl chitosan chloride; a literature study on recombinant human growth hormone (somatropin); a literature study on nasal drug administration; formulation of a suitable Pheroid carrier; entrapment of somatropin in the Pheroid carrier, and in vivo evaluation of nasal absorption of somatropin in Sprague-Dawley rats. / Thesis (M.Sc. (Pharmaceutics))--North-West University, Potchefstroom Campus, 2007.

CORTEZ, S.A.E. O efeito do trabalho infantil na estatura final de adultos jovens e características de sua escolaridade - Estudo da coorte de nascidos vivos entre 1978/79, nos hospitais de Ribeirão Preto, SP. 2005. 158 f. Tese (Doutorado) Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, 2005. Justificativa: Efeitos negativos a saúde da criança têm sido associados ao trabalho infantil. Entretanto, seus efeitos sobre o desenvolvimento humano ainda precisam ser melhor avaliados em razão do pequeno número de estudos existentes da deficiência de controle de fatores socioeconômicos. Objetivo: avaliar o efeito do trabalho infantil na estatura final e algumas características da escolaridade de jovens entre 22-25 anos pertencentes à coorte de nascidos vivos de parto único hospitalar de Ribeirão Preto, SP - 1978/79. Métodos: O presente estudo incluiu 2063 jovens, pertencentes a coorte inicial de 6827 nascidos vivos na cidade de Ribeirão Preto, Brasil (1978/79). Foram seguidos 30,2% dos homens e 33,5% das mulheres. O trabalho infantil foi classificado de acordo com a idade de início do primeiro trabalho: < 14, 14-16 e 17+ anos. Determinantes da altura foram considerados variáveis de confusão na análise, ao nascer (restrição do crescimento intra-uterino RCIU; comprimento ao nascer, idade materna, escolaridade materna, hábito de fumar na gravidez e ocupação do chefe de família) e na vida adulta (cor/etnia, número de irmãos e atividade física). Para as mulheres, idade da menarca também foi considerada. Análise de regressão linear múltipla estratificada por sexo foi realizada após análise bivariada. As variáveis foram selecionadas para o modelo utilizando-se o procedimento passo a passo com seleção retrógrada das variáveis. Interações plausíveis foram testadas. Análise seqüencial dos fatores associados à altura final também foi realizada, separadamente por sexo. Resultados: A altura média foi 176,0 cm (IC 95% 175,6 176,4) para homens e 162,7 cm (IC 95% 162,3 - 163,0) para mulheres. Trabalho antes dos 14 anos de idade foi observado em 20,4% dos homens e 12,4% das mulheres e trabalho entre 14 16 anos foi observado em 41,7 % dos homens e 36,7% das mulheres. Na análise bivariada o trabalho infantil foi associado à mais baixa estatura final tanto para homens (p= 0.007) quanto para mulheres (p=0.004). No entanto, esta associação não se manteve após o controle pelos fatores de confusão. A análise seqüencial sugere que tanto para os meninos como para as meninas as variáveis sócio econômicas interferiram na associação do trabalho infantil e altura final. Ter trabalhado na infância resultou em pior resultado escolar comparados com quem não trabalhou na infância. Aqueles que iniciaram no trabalho antes dos 14 anos e que não estudavam no momento da pesquisa, 62,3% pararam seus estudos no ensino médio sendo que 30,2% pararam de estudar antes de completar 9 anos de estudo. Para quem iniciou o trabalho entre 14-17 anos, 66% chegaram ao ensino médio e 16,4% tem escolaridade acima de 12 anos. Conclusão: Os resultados deste estudo não corroboram com a hipótese de associação independente entre trabalho infantil e altura final. Este resultado se explica pela ação dos confundidores sociais. Observou-se também que não ter trabalhado na infância propicia um melhor resultado escolar, sendo observado que 60,2 % dos que iniciaram atividade laborativa após 17 anos tem escolaridade alta (acima de 12 anos). / CORTEZ, S.A.E. The effects of child labor on the final height and schooling level of a 22-25 years old population-based-cohort from Ribeirão Preto, Brazil. 2005. 158 f. Thesis (Doctoral) Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, 2005. Background: Several adverse health effects have been associated with child labor. However, its effects on human growth are still debatable mostly because of the poor study designs and lack of control of socioeconomic factors. Aim: to assess the effects of child labor on the final height and schooling level of a 22-25-yr old population-based-cohort from Ribeirão Preto, State of São Paulo, located in one of the wealthiest areas in Brazil.Methods: The study included 2063 singletons (30,2% males and 33,5% females), from the initial cohort of 6827 live births in the city of Ribeirão Preto, Brazil (1978/79). Height was measured in centimeters at the follow-up examination. Child labor was defined according to the age at the first job (< 14; 14-16; 17yrs+). Known determinants of height and possible confounders were considered in the analysis, at birth (intrauterine growth restriction IUGR, length at birth, mothers age and education, maternal smoking habit, and socioeconomic position), and at adult age (color/ethnicity, number of siblings, physical activity). For females, age at menarche was also considered as confounder. Statistical analysis was performed separately for males and females. Bivariate and stratified analysis was followed by multivariate linear regression analysis. Variables were selected into the model using a backward stepwise-like selection. Plausible interactions were tested and residual analysis was performed. Moreover, sequential analysis of confounder factors associated to the final height was performed. Results: Mean height was 176.0 cm (95% CI 175.6-176.4) for males and 162.7 cm (95% CI 162.3 163.0) for females. Labor before 14 yrs of age was observed in 20.4% of males and 12.4% of females, and labor between 14 and 16 yrs of age was observed for 41.7% of males and 36.7% of females. In the bivariate analysis child labor was statistically associated with lower height for both males (p=0.0065) and females (p=0.007). However, this association did not remain significant after adjusting for confounders. The sequential analysis suggested that for both males and females socioeconomic factors were the most important factors. Regarding schooling achievement, among those who worked before 14 years old and were not studying at the moment of the study, 62,3% finished or withdraw the studies at any moment during the high school the 30.2% before completing 9 years of study. High school profile was similar for those who begin to work between 14 and 17 years old and 16.4% achieved more than 12 years old of study. Conclusion: our study did not support the association between child labor and final height. Moreover, it was observed that those children who did not work early in life achieved higher schooling levels.

Ng, Siu Fung. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 206-211). / Abstracts in English and Chinese. / Acknowledgements --- p.i / Abstract --- p.iii / 摘要 --- p.v / Table of contents --- p.vii / List of figures --- p.xv / List of tables --- p.xix / List of abbreviations --- p.xx / Chapter / Chapter 1. --- Introduction / Chapter 1.1 --- Growth hormone --- p.1 / Chapter 1.1.1 --- Historic discovery of growth hormone --- p.1 / Chapter 1.1.2 --- Structural and functional study of GH --- p.1 / Chapter 1.1.2.1 --- Molecular evolution of GH --- p.1 / Chapter 1.1.2.2 --- Two-dimensional and three dimensional structures --- p.5 / Chapter 1.1.2.3 --- Heterogeneity of GH --- p.8 / Chapter 1.1.2.4 --- Regulation and secretion pattern of GH --- p.9 / Chapter 1.1.2.5 --- Circulation of GH in blood --- p.11 / Chapter 1.1.2.6 --- Biological activity of GH in human --- p.12 / Chapter 1.2 --- GH receptor and signal transduction --- p.12 / Chapter 1.3 --- GH disorder --- p.15 / Chapter 1.4 --- Treatment for GH disorder --- p.16 / Chapter 1.5 --- GH assay --- p.17 / Chapter 1.6 --- Aims of study --- p.19 / Chapter 2. --- SUMO-hGH expression vector construction / Chapter 2.1 --- Introduction --- p.21 / Chapter 2.2 --- Fusion partner - SUMO --- p.23 / Chapter 2.3 --- Materials --- p.24 / Chapter 2.3.1 --- Reagents for bacterial culture --- p.24 / Chapter 2.3.2 --- Reagents for agarose gel electrophoresis --- p.26 / Chapter 2.3.3 --- 2'-deoxyribonucleoside 5'-triphosphate mix for polymerase chain reaction --- p.26 / Chapter 2.3.4 --- Sonication buffer --- p.26 / Chapter 2.3.5 --- Modified solubilization buffer --- p.27 / Chapter 2.3.6 --- Reagents for sodium dodecylsulphate polyacrylamide gel electrophoresis --- p.27 / Chapter 2.4 --- Methods --- p.29 / Chapter 2.4.1 --- General techniques in molecular cloning of hGH gene --- p.29 / Chapter 2.4.2 --- Expression of SUMO-hGH fusion protein - small scale --- p.42 / Chapter 2.4.3 --- General protein analysis --- p.43 / Chapter 2.5 --- Results --- p.45 / Chapter 2.5.1 --- Molecular cloning of hGH gene into expression vector --- p.45 / Chapter 2.5.2 --- Expression of SUMO-hGH --- p.46 / Chapter 2.5.3 --- Modification of the expression conditions --- p.46 / Chapter 2.6 --- Discussion --- p.50 / Chapter 2.6.1 --- Expression vector --- p.53 / Chapter 2.6.2 --- Protein expression --- p.53 / Chapter 2.7 --- Conclusion --- p.54 / Chapter 3. --- SUMO-hGH purification and downstream processing / Chapter 3.1 --- Introduction --- p.55 / Chapter 3.2 --- Immobilized-metal affinity chromatography --- p.55 / Chapter 3.3 --- SUMO protease --- p.57 / Chapter 3.4 --- Materials --- p.59 / Chapter 3.4.1 --- Reagents for IMAC purification of SUMO-hGH fusion protein --- p.59 / Chapter 3.4.2 --- Reagents for IMAC purification of mature rhGH --- p.60 / Chapter 3.4.3 --- Reagents for Western blotting --- p.60 / Chapter 3.4.4 --- Gel filtration running buffer --- p.62 / Chapter 3.5 --- Methods --- p.62 / Chapter 3.5.1 --- Purification of SUMO-hGH fusion protein by Ni2+-NTA affinity chromatography --- p.62 / Chapter 3.5.2 --- Cleavage of His-SUMO fusion partner to generate mature rhGH --- p.63 / Chapter 3.5.3 --- Purification of mature rhGH by 2nd round of Ni2+-NTA affinity chromatography --- p.64 / Chapter 3.5.4 --- Purification of rhGH by size exclusion chromatography - gel filtration chromatography --- p.64 / Chapter 3.5.5 --- General protein analysis --- p.65 / Chapter 3.6 --- Results --- p.67 / Chapter 3.6.1 --- Purification of SUMO-hGH fusion protein by Ni2+-NTA affinity chromatography --- p.67 / Chapter 3.6.2 --- Cleavage of His-SUMO fusion partner to generate mature rhGH --- p.69 / Chapter 3.6.3 --- Digestion efficiency of different constructs of SENP1C --- p.73 / Chapter 3.6.4 --- Purification of mature rhGH by 2nd round of Ni2+-NTA affinity chromatography --- p.77 / Chapter 3.6.5 --- Purification of rhGH by size exclusion chromatography -gel filtration chromatography --- p.78 / Chapter 3.7 --- Discussion --- p.81 / Chapter 3.7.1 --- Purification of SUMO-hGH fusion protein by Ni2+-NTA affinity chromatography --- p.81 / Chapter 3.7.2 --- Cleavage of His-SUMO fusion partner to generate mature rhGH --- p.82 / Chapter 3.7.3 --- Purification of mature rhGH by 2nd round of Ni2+-NTA affinity chromatography --- p.82 / Chapter 3.7.4 --- Purification of rhGH by size exclusion chromatography -gel filtration chromatography --- p.85 / Chapter 3.8 --- Conclusion --- p.85 / Chapter 4. --- Fermentation expression of SUMO-hGH and scale-up of downstream process / Chapter 4.1 --- Introduction --- p.86 / Chapter 4.2 --- Bioreactor system for E.coli host cultivation --- p.87 / Chapter 4.3 --- Mechanical cell disruption for cell --- p.88 / Chapter 4.4 --- rhGH binding assay --- p.88 / Chapter 4.5 --- Materials --- p.89 / Chapter 4.5.1 --- Reagents for bacterial culture by fermenter --- p.89 / Chapter 4.5.2 --- Reagents for HEK293 Hi cultivation --- p.91 / Chapter 4.5.3 --- Reagents for Dual-Luciferase® Reporter Assay System --- p.92 / Chapter 4.5.4 --- Reagents for silver stain of SDS-PAGE mini-gel --- p.93 / Chapter 4.6 --- Methods --- p.94 / Chapter 4.6.1 --- Bioreactor system and fixed volume fed-batch fermentation --- p.94 / Chapter 4.6.2 --- Large scale mechanically disruption of cell membrane --- p.97 / Chapter 4.6.3 --- Downstream processing of SUMO-hGH --- p.97 / Chapter 4.6.4 --- Culture of HEK293 Hi cells --- p.97 / Chapter 4.6.5 --- Dual-Luciferase® Reporter Assay System --- p.98 / Chapter 4.6.6 --- Silver staining of SDS-PAGE mini-gels --- p.101 / Chapter 4.7 --- Results --- p.101 / Chapter 4.7.1 --- Fed-batch fermentation of E. coli BL21 --- p.101 / Chapter 4.7.2 --- Comparison on disruption methods and the purification of SUMO-hGH from cell lysate --- p.106 / Chapter 4.7.3 --- Optimization of His-MBP-SENP1C digestion condition --- p.108 / Chapter 4.7.4 --- Optimization of rhGH purification in 2nd round of IMAC --- p.110 / Chapter 4.7.5 --- Characterization of mature rhGH --- p.112 / Chapter 4.8 --- Discussion --- p.116 / Chapter 4.8.1 --- Fed-batch fermentation of E. coli BL21 --- p.118 / Chapter 4.8.2 --- Downstream processing of fermentation culture and characterization of rhGH --- p.120 / Chapter 4.8.3 --- M9 based defined medium fermentation study --- p.122 / Chapter 4.8.4 --- rhGH production yield estimation --- p.128 / Chapter 4.8.5 --- Comparison of our fermentation expression system to the published data --- p.130 / Chapter 4.9 --- Conclusion --- p.132 / Chapter 5. --- His-MBP-SENPIC expression and purification / Chapter 5.1 --- Introduction --- p.133 / Chapter 5.2 --- Materials --- p.134 / Chapter 5.2.1 --- Reagents for bacterial culture --- p.134 / Chapter 5.2.2 --- Reagents for immobilized metal affinity chromatography purification of His-MBP-SENP1C --- p.135 / Chapter 5.3 --- Methods --- p.136 / Chapter 5.3.1 --- Expression of His-MBP-SENP1C --- p.136 / Chapter 5.3.2 --- Semi-purification of His-MBP-SENP1C by Ni2+-NTA affinity chromatography --- p.138 / Chapter 5.4 --- Results --- p.139 / Chapter 5.4.1 --- Expression of His-MBP-SENP1C --- p.139 / Chapter 5.4.2 --- Digestion activity of His-MBP-SENP1C expressed --- p.139 / Chapter 5.5 --- Discussion --- p.141 / Chapter 5.5.1 --- Expression and purification of His-MBP-SENP1C --- p.141 / Chapter 5.5.2 --- His-MBP-SENP1C production yield estimation --- p.143 / Chapter 6. --- Production and characterization of monoclonal antibodies against rhGH / Chapter 6.1 --- Introduction --- p.145 / Chapter 6.2 --- Materials --- p.146 / Chapter 6.2.1 --- Reagents for Sp2/0-Ag14 cultivation --- p.146 / Chapter 6.2.2 --- Reagents for PEG fusion --- p.147 / Chapter 6.2.3 --- Reagents for enzyme linked immunosorbent assay --- p.149 / Chapter 6.2.4 --- Reagents for mAbs purification by HiTrap´ёØ Protein G HP Column --- p.150 / Chapter 6.3 --- Methods --- p.151 / Chapter 6.3.1 --- ELISA --- p.151 / Chapter 6.3.2 --- Immunization --- p.152 / Chapter 6.3.3 --- Culturing of myeloma fusion partner cells --- p.153 / Chapter 6.3.4 --- Isolation of splenocyte --- p.153 / Chapter 6.3.5 --- PEG fusion --- p.154 / Chapter 6.3.6 --- Limiting dilution --- p.155 / Chapter 6.3.7 --- Cryopreservation of hybridoma cell lines --- p.156 / Chapter 6.3.8 --- Mass production of monoclonal antibodies --- p.157 / Chapter 6.3.9 --- Purification of IgG mAbs from ascites --- p.157 / Chapter 6.3.10 --- MAbs isotyping --- p.159 / Chapter 6.3.11 --- Determination of kinetic parameters of mAbs --- p.159 / Chapter 6.4 --- Results --- p.162 / Chapter 6.4.1 --- Production of murine anti-rhGH monoclonal antibodies --- p.162 / Chapter 6.4.2 --- Characterization of anti-rhGH mAbs --- p.170 / Chapter 6.5 --- Discussion --- p.178 / Chapter 6.5.1 --- Mass production of mAbs --- p.179 / Chapter 6.5.2 --- Future works on mAbs --- p.179 / Chapter 6.6 --- Conclusion --- p.181 / Chapter 7. --- Development of sandwich ELISA for rhGH / Chapter 7.1 --- Introduction --- p.182 / Chapter 7.2 --- Materials --- p.184 / Chapter 7.2.1 --- Reagents for sandwich ELISA --- p.184 / Chapter 7.3 --- Methods --- p.184 / Chapter 7.3.1 --- Production of rabbit polyclonal antiserum against rhGH --- p.184 / Chapter 7.3.2 --- Sandwich ELISA --- p.185 / Chapter 7.4 --- Results --- p.186 / Chapter 7.4.1 --- Production of rabbit antiserum against rhGH --- p.186 / Chapter 7.4.2 --- Sandwich ELISA --- p.188 / Chapter 7.4.3 --- Optimization of sandwich ELISA --- p.190 / Chapter 7.4.4 --- Specificity of sandwich ELISA --- p.194 / Chapter 7.4.5 --- Cross reactivity of sandwich ELISA to E.coli cell lysate --- p.196 / Chapter 7.4.6 --- Measurement of SUMO-hGH with sandwich ELISA --- p.198 / Chapter 7.5 --- Discussion --- p.201 / Chapter 7.5.1 --- Application of sandwich ELISA --- p.203 / Chapter 7.5.2 --- Future works on sandwich ELISA --- p.205 / Chapter 7.6 --- Conclusion --- p.205 / References --- p.206 / Appendix - pJ2:G01458 nucleotide sequence --- p.213

Thesis (M. Sc. (Statistics)) -- University of Limpopo, 2018 / TheEllisrasLongitudinalStudy(ELS)openedthepossibilitiesforunderstandingthegrowthvariationsamongchildreninruralSouthAfrica. Theaimofthe study was to analyse the growth spurt in height and weight of children using the ELS. This is part of the on going ELS and this study followed secondary analytical longitudinal study using data collected from November 1996 to November 2003. All children underwent a series of anthropometric measurements of height and weight according to the standard procedures recommended by the International Society for the Advancement of Kinanthropometry. The descriptive statistics was done for age, height, weight, velocity and acceleration by gender amongst rural children in Ellisras. Thelinearmixedmodelwasusedto analyse data. Based on the smallest values of AIC and BIC, the best model to fit the ELS data which was found to be the unstructured covariance structure model was chosen. The interaction between age and gender, which was significant at 5% level suggested that the relationship of age with growth varies depending on gender. There was also a significant positive linear relationship of age with distance. The onset of growth spurt for rural children in Ellisras was at 12.05 years for boys and at 12.32 years for girls, while the Senegalese boys took off earlier at 11.02 years. Ellisras rural boys and USA boys had their onset of growth spurt almost at the same age at 13 years for USA boys and 12.05 years for Ellisras rural boys. USA girls had their onset of growth spurt earlier at 11 years than Ellisras rural girls at 12.32 years. Newcastle upon Tyne adolescents reached ii their PHV at 14 and 12 years for boys and girls respectively, similarly with Ellisras rural children at 14.21 and 11.80 years for boys and girls respectively. Ellisras rural girls had their PHV at 11.80 years earlier than Ellisras rural boys at 14.21 years. Children in rural Ellisras in the ELS and their growth variations do not differ that much compared with other children across the world. Key words: Growth spurt, peak height velocity, boys, girls. / National Research Foundation (NRF)

In this study, the effects of bioprocess operation parameters on recombinant human growth hormone (rhGH) production by P. pastoris were systematically investigated. In this frame, first, for the extracellular expression and purification of human growth hormone by recombinant P. pastoris the cDNA of hGH, fused with a polyhistidine tag and also fused with a target site for the Factor Xa protease in which cleavage produces a mature N- and C- termini of rhGH, was cloned into pPICZ&amp / #945 / A plasmid and the constructed system within the plasmid, pPICZ&amp / #945 / A::hGH, was integrated to AOX1 locus of P. pastoris and expressed under alcohol oxidase promoter which is induced by methanol. With dot-blot analysis, the appropriate two strains producing human growth hormone at high levels and having different methanol utilization phenotype (Mut+ and Muts) were chosen among the other transformants. Then, the effects of methanol concentrations on the expression of rhGH and cell growth were analyzed and both of the phenotypes were compared in defined and complex media in laboratory scale air filtered shake bioreactors. The highest rhGH concentration for Mut+ and MutS, was found as 0.052 kg m-3 and 0.16 kg m-3, respectively, at 2 %(v/v) methanol concentration in complex medium. When methanol was used as the sole carbon source in defined medium, Muts phenotype had very low specific growth rate on methanol due to the intrinsic characteristics of it, therefore detectable rhGH was not observed, on the other hand, optimum rhGH concentration produced by Mut+ strain was found as 0.032 kg m-3 at 3% (v/v) methanol concentration in defined medium. In mixed system (glycerol/methanol) which is also defined, when the optimum glycerol concentration, 30 kg m-3, was used, Muts produced the highest rhGH, 0.110 kg m-3, at 1% (v/v) methanol concentration and any increase in methanol concentration resulted in lower rhGH production, on the other hand, Mut+ strain produced 0.060 kg m-3 rhGH at 4% (v/v) methanol concentration, which indicated that higher rhGH production capacity of Mut+ strain was obtained at high methanol concentrations. Using the designed defined medium for Mut+ phenotype where methanol was used as the sole carbon source with an optimum concentration of 3% (v/v), the effects of oxygen transfer on rhGH production, by-product formation, and cell growth, oxygen transfer and fermentation characteristics were investigated by using pilot scale bioreactor. Oxygen transfer effects on rhGH production were investigated at QO/VR=0.5 vvm / N=250, 500, 625, 750 min-1 conditions. The variations in rhGH , cell, amino acid and organic acid concentrations with the cell cultivation time, specific cell growth rate, the oxygen uptake rate, the liquid phase coefficient by using the dynamic method, maintenance coefficient for oxygen and yield coefficients were determined. The highest rhGH concentration was obtained at 0.5 vvm, 500 min-1 condition as 0.023 kg m-3 with 5.37 kg m-3 cell density.

The purpose of the study is to purify human growth hormone from the fermentation broth by affinity chromatography. For this purpose, human growth hormone specific oligonucleotide aptamers are selected among an aptamer library / selected oligonucleotides were synthesized and used as ligands. Effect of pH on ligand-human growth hormone complex formation was investigated and the highest complex formation was obtained at pH= 7.0. Human growth hormone is separated from the fermentation broth with 99.8% purity and 41% overall yield. The equilibrium data obtained was described by Langmuir type isotherm where saturation constant (q0) and affinity constant (K) are calculated as 0.338 mg hGH/&igrave / mol aptamer and 0.059 mg hGH/ml, respectively. Further, equilibriumdata obtained using aptamer affinity column was described by Langmuir type isotherm where saturation constant (q0) and affinity constant (K) are 0.027 mg hGH/&igrave / mol aptamer and 1.543 mg hGH/ml, respectively. It is possible that, selected aptamer can be used for purification of bulk amounts of recombinant human growth hormone by using aptamer affinity chromatography.