O presente estudo teve por objetivo avaliar protocolo de reabilitação para o período pós-operatório inicial de artroscopias visando diminuir a inflamação no local operado e aumentar a mobilidade articular. Foram utilizados 12 equinos (total de 20 articulações) encaminhados para artroscopia com diagnóstico de osteocondrite dissecante. Dessas, dez articulações receberam protocolo de reabilitação nos primeiros cinco dias do período pós-operatório. O protocolo consistiu em crioterapia, movimentação passiva da articulação e exercício controlado de baixa intensidade, além de uso sistêmico de anti-inflamatório. O outro grupo, também composto por dez articulações, recebeu apenas a terapia utilizada rotineiramente no HOVET-USP, consistido de repouso em baia e antiinflamatório. As articulações foram avaliadas quanto à circunferência em centímetros, ângulo de flexão, termografia, grau de claudicação. Amostras de líquido sinovial foram coletadas imediatamente antes do procedimento cirúrgico (D1), após 48h (D3) e após 96h (D5) para análise física, qualidade do coágulo de mucina, e quantificação de biomarcadores (IL-1, IL-6 e IL-10, PGE2 e SAA). As análises de exame de claudicação, circunferência articular, ângulo de flexão articular e termografia não apresentaram diferenças significativas entre os grupos, nem entre os diferentes dias do mesmo grupo. Na análise do líquido sinovial, a cor e o aspecto apresentaram piora do D1 para o D3, de amarelo claro para avermelhado e de límpido para turvo, respectivamente, nos dois grupos. No entanto, no grupo tratado houve melhora do D3 para o D5, tanto para cor (de avermelhado para maioria xantocrômica e amarela) como aspecto (de maioria turva para ligeiramente turva). No grupo controle os líquidos permaneceram sem alteração em cor e aspecto de D3 para D5, e nas comparações entre os grupos não houve diferença para D1, D3 e D5. A viscosidade do líquido sinovial no grupo controle diminuiu significativamente quando comparados D1, D3 e D5. Já no grupo tratado a diminuição da viscosidade só foi observada quando comparados D1 e D5. O coágulo de mucina apresentou piora de D1 para D3 no grupo controle, com elevação não significativa de D3 para D5, enquanto que para o grupo tratado não houve diferença significativa de D1 para D3 e de D3 para D5, quando comparados o D5 dos dois grupos, o tratado obteve melhor qualidade. As concentrações de interleucina nas amostras não forneceram dados suficientes para análise. Na análise das concentrações de PGE2 não houve diferença entre os grupos nos diferentes momentos, ocorrendo elevação de D3 para D5 em ambos os grupos, porém, no grupo tratado não há diferença entre D1 e D5. Já para SAA os grupos apresentaram comportamento similar de resposta, com elevação de D1 para D3 e queda de D3 para D5, porém menos acentuado no grupo tratado, o que levou a diferença entre os grupos em D3. Pode-se concluir, que o protocolo de reabilitação, apesar de não gerar diferença significativa para as avaliações de exame físico dos animais, proporcionou melhor qualidade de líquido sinovial quanto a cor, aspecto, viscosidade e precipitado de mucina, além de evidenciar menores elevações nas concentrações de marcadores inflamatórios no liquido sinovial durante o período estudado. / The purpose of this study was to evaluate a rehabilitation protocol for the initial postoperative period of metatarsophalangeal, metacarpophalangeal and tarsocrural´s arthroscopies, which seeks to, minimize local inflammation, diminish swelling, promote better joint range of motion and pain relief during such period. Twelve horses participated in this study - amounting to 20 joints - with dissecans ostheochondritis diagnosis. The first group was formed by ten joints, which were treated under rehabilitation protocol for the first 5 days as from the surgery (Treated group). The rehabilitation protocol consisted of cryotherapy, passive range of motion, low intensity exercise and non-steroidal anti-inflammatory drug. The second group also formed of ten joints received the standard HOVET-USP therapy, which consists of rest and non-steroidal anti-inflammatory drug Both groups were treated with the same non-steroidal anti-inflammatory drugs. The joints were measured for circumference, maximal flexion angle, thermography, and lameness score on the day before the surgery (D0) and during the first four days after the surgery. Synovial fluid samples were collected immediately before surgery (D1), within 48 hours (D3), and within 96 hours from the surgery (D5). The analysis evaluated gross appearance (color and aspect), viscosity and mucin clot quality, as well as biomarkers (Il-1, Il-6, Il- 10, PGE2, and SAA) quantification. Lameness examination, joint circumference, flexion angle and thermography evaluation were not significantly different between groups. In synovial fluid analyses de color and aspect have worsen from D1 (clear light yellow) to D3 (turbid hemorrhagic) in both groups. On treated group color and aspect improved from D3 (turbid hemorrhagic) to D5 (xanthochromic and yellow slightly turbid). On treated group there was no difference between D3 and D5. When the groups were compared, none significant differences was seen. The fluid viscosity of control group had significant decrease from D1, to D3 and from D1 and D5. In treated group this viscosity decrease was only seen between D1 and D5. The mucin clot formation worsened when D1 e D3 of control group was compared and remains similar from D3 to D5. In treatment group there were no differences when compared D1 with D3 and D3 with D5. The comparison between groups of D5 has shown treated group improved clot. The interleukin couldn´t be measured on sufficient number of samples for the statistics method. There were no differences between groups on all moments. The PGE2 response was similar in both group with a rise on concentration from D3 to D5. In treated group D1 was similar to D5. This results suggests more evident inflammatory response in the control group. For the SAA the groups have shown similar responses, with an increase from D1 to D3 and decrease from D3 to D5. The response on treated group was less intense and demonstrates lower values in D3 when compared with D3 control group. It was concluded with this study that rehabilitation protocol improved synovial fluid analyses for, color, aspect, viscosity and mucin clot. It even had promoted lower concentrations of inflammatory biomarkers for the treated group during the period.

Introdução: Os meningiomas são neoplasias intracranianas de crescimento lento que se originam das células meningoteliais da aracnoide e representam os tumores intracranianos mais comuns, contabilizando 13-26% deste total, sendo um dos primeiros tumores sólidos a terem alterações genéticas identificadas. Inúmeros tem sido os avanços para a melhor compreensão das vias moleculares correlacionadas com a tumorigênese e progressão tumoral dos meningiomas, neste contexto tem se destacado o papel dos microRNAs que são RNAs não-codificantes (ncRNAs) constituídos por 19 a 25 nucleotídeos, cuja função é o silenciamento do RNAm em nível póstranscricional. Portanto, o objetivo do nosso estudo foi avaliar a expressão tecidual e plasmática dos miRNAs miR-181d, miR-181c e miR-130a. Pacientes e métodos: Os miRNAs miR-181d, miR-181c e miR-130a foram selecionados a partir de estudo prévio do nosso grupo pela técnica de análise em larga escala de microarrays, onde foram comparados meningiomas grau I com amostras controles de aracnóides. Neste trabalho foi avaliada expressão destes miRNAs no tecido tumoral e plasma de meningiomas grau I, II e III. Resultados: O miR-181d apresentou-se hiperexpresso nos grupos estudados, no tecido tumoral quanto no plasma. O nível de expressão foi maior de acordo com a progressão do grau do tumor. Os miR-181c e miR-130a não apresentaram diferença estatística nos grupos estudados em ambos tecido tumoral e plasma. Conclusões: O miR-181d tem potencial para ser utilizado como biomarcador para meningiomas e está associado com sua progressão tumoral. / Introduction: Meningiomas are intracranial tumors of slow growth that originate from meningothelial arachnoid cells and represents the most common intracranial tumors, accounting for 13-26% of this total, beeing one of the first solid tumors to have identified genetic alterations There are technological advances available to a better understanding of the molecular pathways correlated with tumorigenesis and tumor progression of meningiomas. The role of microRNAs in this process is very importante. MicroRNAs are non-coding RNAs (ncRNAs) consisting of 19 to 25 nucleotides, with function of mRNA silencing post-transcriptional level. The aim of our study was to evaluate the tissue expression and plasma of miRNAs miR-181d, miR-181c and miR-130a. Patients and methods: The miRNAs miR-181d, miR-181c and miR-130a were selected from a previous study of our group by analysis technique on large scale called microarrays, which were compared meningiomas grade I with arachnoid controls samples. In this study, we evaluated expression of these miRNAs in tumor tissue and plasma meningiomas grade I, II and III. Results: The miR-181d was presented upregulated in the all groups in both tumor tissue and in plasma. The level of expression was increased according to the progression of tumor grade. The miR-181c and miR-130a showed no statistical difference in the groups studied in both tumor tissue and plasma. Conclusions: The miR-181d has potential as a biomarker for meningiomas and is associated with tumor progression.

Orientadores: Luzia Koike, Eugenio Vaz dos Santos Neto / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Química / Made available in DSpace on 2018-08-19T14:25:27Z (GMT). No. of bitstreams: 1 Fontes_RosaneAlves_D.pdf: 10963624 bytes, checksum: 8162d4b98904d9afe273a093366e9221 (MD5) Previous issue date: 2011 / Neste trabalho foram sintetizados quatro biomarcadores ácidos, a saber: ácido 5b (H) colânico (2), 5a (H) homopregnânico (4), 8,13-tetraidrocopálico (6) e 15-isocopálico (7). Para tanto, foram utilizadas matérias primas comerciais (ácido litocólico e estigmasterol) e o ácido copálico (extraído e purificado a partir de bálsamo de copaíba). Também foi preparado o biomarcador hidrocarboneto 5b (H) colano (8). Estas moléculas foram sintetizadas com o objetivo de serem utilizadas como padrões de referência na quantificação de biomarcadores ácidos e hidrocarbonetos presentes em dois conjuntos de óleos provenientes da Bacia Potiguar. O conjunto I contém cinco óleos de diferentes origens, sendo dois lacustres (LC-1 e LC-2), um marinho evaporítico (ME) e dois mistos (M-1 e M-2) e o conjunto II contém quatro óleos de mesma evolução térmica e mesma origem (ME-a, ME-b, ME-c e ME-d), todos marinho evaporíticos. O que os distingue são os diferentes níveis de biodegradação. Utilizou-se também o isocopalano (9), previamente preparado e cedido pelo Dr. Imamura (2011), na quantificação de biomarcadores terpanos. Os resultados das análises quantitativas dos biomarcadores terpanos e esteranos, quantificados com os padrões de referência 9 e 8, respectivamente, mostraram que quando se utiliza somente o 8 como padrão referência para quantificação de todos os biomarcadores saturados, a quantidade de hopanos fica subestimada em quase 60% quando comparada a quantificação dos mesmos utilizando-se 9. No entanto, isto não significa que este seja um padrão mais adequado a quantificação de todos os terpanos. Quanto às análises dos extratos ácidos de petróleo utilizando-se a técnica de espectrometria de massas seqüencial Trap Linear-Quadrupolo/Orbitrap (LTQ-Orbitrap), observou-se que os resultados foram satisfatórios contribuindo com informações qualitativas úteis para distinção de origem e biodegradação quando relacionadas às distribuições dos ácidos carboxílicos, de acordo com o grau de insaturação destas moléculas. Os perfis moleculares encontrados para os extratos ácidos são muito similares aos que se obtinham com os métodos convencionais de análise de ácidos carboxílicos de petróleo, quando convertidos a ésteres metílicos ou reduzidos a hidrocarbonetos e analisados por CG-EM. A principal vantagem da utilização do Orbitrap na análise qualitativa de ácidos carboxílicos está na redução drástica do tempo de análise atrelada à dispensa de reações de derivatização. Contudo, apesar da altíssima resolução de massas (100.000) o Orbitrap não é capaz de distinguir íons moleculares de estereoisômeros, os quais são altamente difundidos entre os biomarcadores de petróleo. Deste modo, no presente trabalho não foi possível realizar as análises quantitativas dos biomarcadores ácidos. Evidentemente, as técnicas convencionais de análise de biomarcadores ácidos, como CG-EM, permanecem altamente relevantes para estudos geoquímicos. Com relação aos estudos realizados de EM com os padrões de biomarcadores ácidos, sintetizados neste trabalho, foi observado que existe uma correlação direta entre as suas estruturas químicas e os fragmentos gerados a partir da dissociação induzida por colisão dos mesmos. Isto revela o grande potencial desta técnica para obtenção de fragmentos disgnósticos de famílias de biomarcadores ácidos presentes em petróleo a partir da análise direta de extratos ácidos / Four acidic biomarkers were synthesized in this work, namely: 5 b (H) Cholanic acid (2), 5a (H) homopregnanic acid (4), 8, 13-tetrahydrocopalic acid (6) and 15-isocopalic acid (7). For this purpose, were used commercial raw materials (lithocolic acid and stigmasterol) and copalic acid (extracted and purified from copaiba balsam). It was also assembled a hydrocarbon biomaker 5 b (H) Colane (8). These molecules were synthesized in order to be used as reference standards in the quantification of acids and hydrocarbons biomarkers present in two sets of oils from the Potiguar Basin. The set I contains five oils from different sources, two lacustrine (LC-1 and LC-2), a marine evaporitic (ME) and two mixed (M-1 and M-2) and set II contains four oils from the same source (ME-a, ME-b, ME-c and ME-d), all marine evaporitic and the same thermal evolution. What distinguishes them are the biodegradation levels. We also used the isocopalane (9), previously prepared and given by Imamura (2011) in the quantification of terpanes biomarkers. The results of quantitative analysis of terpanes and steranes biomarkers, quantified with reference standards 9 and 8, respectively, showed that when using only the 8 as the reference standard for quantification of all saturated biomarkers, the amount of hopanes is underestimated by almost 60 % when compared to quantification using 9. However, this does not mean that it is a more appropriate standard to quantify all terpanes. Concerning acidic biomarkers analysis using sequential mass spectrometry Linear-Quadrupole/Orbitrap (LTQ-Orbitrap) the results were satisfactory, contributing with qualitative useful information for distinction of origin and the of biodegradation levels. Results are very similar to that obtained with conventional methods of analysis of carboxylic acids from oil, when converted to methyl esters, or reduced to hydrocarbons and analyzed by GC-MS. The main advantage of using the Orbitrap, in the qualitative analysis of acids biomarkers, is the drastic reduction in the time consumed of analysis linked to absence of derivatization reactions. However, despite the very high mass resolution (100,000), Orbitrap is not able to distinguish molecular stereoisomers, which are highly disseminated among the biomarkers of oil. Thus, it was impossible to carry out the quantitative analysis of acidic biomarkers. Of course, conventional techniques of analyses of acidic biomarkers, such as GC-MS, remain highly relevant to geochemical studies. With regard to studies of MS with standard acidic biomarkers, synthesized in this work, the results showed that there is a direct correlation between their chemical structures and fragments generated from the collision induced dissociation. This shows the great potential of this technique to obtain diagnostic fragments of biomarker families present in oils from the direct analysis of acidic extracts / Doutorado / Quimica Organica / Doutor em Ciências

Orientador: Orival Andries Júnior / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Educação Física / Made available in DSpace on 2018-08-20T08:53:03Z (GMT). No. of bitstreams: 1 Moraes_RafaelCarvalhode_D.pdf: 3074022 bytes, checksum: 193ac376aa88ed39b3128919ba4f7299 (MD5) Previous issue date: 2012 / Resumo: Em se tratando de treinamento esportivo, é possível afirmar que gestos técnicos e táticos são bastante explorados e também aplicados às sessões de treino. Contudo, em relação às respostas e adaptações fisiológicas, ainda há muitos fatores que não recebem a devida importância. O objetivo deste trabalho foi analisar biomarcadores sanguíneos, Escala de Borg (EB) (dor e esforço) e frequência cardíaca (FC) em estímulos anaeróbios máximos no nado crawl (25, 50, 100 e 200 metros). Assim, 4 nadadores (sexo masculino) competitivos em nível nacional, com idade média de 20 anos (± 2,71), treinando há no mínimo 3 anos, foram voluntários no estudo. Os atletas seguiram uma rotina de treino em água de 2hs diárias - cinco a seis vezes por semana - durante 48 semanas, minuciosamente controladas, objetivando 3 competições-alvo, especialmente da 46ª semana deste período. Durante as 10 semanas precedentes à competição principal cada atleta realizou 4 estímulos máximos nas metragens supracitadas em 3 momentos - semanas 1 e 2 (momento 1), 5 e 6 (momento 2) e 9 e 10 (momento 3), mantendo a rotina de treino proposta. Foram coletadas amostras de sangue para medir as concentrações de lactato, creatinina, creatina quinase (CK) e ácido úrico (AU). Além disso, foram aferidos os tempos de cada estímulo, velocidade, FC e EB. As amostras de sangue, dados de FC e EB, foram coletadas antes e em momentos pós-testes. Para análise estatística utilizou-se do teste Kruskal-Wallis para medidas não-paramétricas. Os valores para os biomarcadores sanguíneos não apresentaram diferenças significativas entre os momentos de avaliação (p<0,05). A FC apresentou diferenças entre 25, 50 e 100m de cada teste, e apresentou valores de correlação com velocidade (r=-0,409), com lactato (r=-0,8) e creatinina (r=0,341). Neste contexto, a EB mostrou-se um bom parâmetro para prescrição da intensidade de dor e esforço quando relacionada a velocidade (R2= 58,9% e R2=74,6% respectivamente), lactato (R2= 48,5% e R2= 44,3% respectivamente) e FC (R2= 28,9% e R2= 23,8% respectivamente). Assim, CK, creatinina, AU e FC não mostraram-se bons parâmetros para diferenciação de esforços nos estímulos analisados. Desta forma, adotar valores fixos de FC e/ou algum biomarcador sanguíneo pode sub ou superestimar o desempenho do atleta. Já o lactato pode ser considerado bom parâmetro avaliador para estímulos anaeróbios máximos sendo mais otimizado quando atrelado à FC / Abstract: In the case about sportive training, it is possible to affirm that technical and tactical gestures are fairly well explored and applied to the training sessions. However, about physiological adaptations there are many factors that still has not been given its due importance. The objective of this study was to analyze blood biomarkers along with the Borg scale (BS) (pain and effort) and heart rate (HR) in the maximum anaerobic bouts in craw swimming (25, 50, 100 and 200 meters). Thus, four swimmers (men) competing at the national level, with an average age of 20 years (± 2.71), training for at least three years, volunteered for the study. The athletes followed a routine of 2hs daily training - five to six times a week - 48 weeks, carefully controlled, aiming at 3 competitions, specially the 46ª week competition of period. In the 10 weeks preceding the main competition, each athlete performed 4 maximum bouts in that distances at 3 times - weeks 1-2 (moment 1), 5-6 (moment 2) and 9-10 (moment 3), keeping the training routine proposal. We collected blood samples to measure the concentrations of lactate, creatinine, creatine kinase (CK) and uric acid (UA). In addition, we measured the time of each bouts, speed, HR and SB. Blood samples, FC and EB data were collected before and after testing times. For statistical analysis we used the Kruskal-Wallis test for nonparametric measures. The values for blood biomarkers showed no significant differences between the moments (p <0.05). The HR differ even between the distances 25, 50 and 100m of each test, and reported correlated values with speed (r=-0,409), lactate (r=-0,8) and creatinine (r=0,341). So, the SB was shown to be a good parameter of pain abd effort for the prescription of exercise intensity when related with speed (R2= 58,9% e R2=74,6% respectively), lactate (R2= 48,5% e R2= 44,3% respectively) and HR (R2= 28,9% e R2= 23,8% respectively). Thus, CK, creatinine, UA and HR, wasn't good parameters to differ the effort between bouts analyzed. Then, adopting fixed values of HR and / or any blood biomarker may under or overestimate the performance of the athlete. The lactate coupled to HR can be treated like a good measure parameter of anaerobic bouts / Doutorado / Ciencias do Desporto / Doutor em Educação Física

La dengue est une maladie virale endémique dans les pays tropicaux. Bénigne dans sa forme classique (FC), elle peut être redoutable lors de complications associées à sa forme sévère (FS). Actuellement, l’établissement du pronostic d’évolution vers une dengue sévère est peu fiable. Afin d’identifier des marqueurs précoces d’évolution vers une forme sévère, une étude de caractérisation différentielle de plasmas FC et FS prélevés précocement, a été effectuée par différentes approches de protéomique. La protéine non-structurale NS1 est une des protéines virale associée à la pathogénicité. Une méthode de profilage des plasma utilisant la technologie SELDI-TOF/MS (Surface Enhanced Laser Desorption Ionization-Time of Flight/Mass Spectrometry) couplée à un anticorps monoclonal anti-NS1 a été utilisée afin d’identifier les ligands de cette protéine. La protéine NS1 a été détectée spécifiquement dans des plasmas de patients dengue. Aucun partenaire de la NS1 n’a pu être identifié. La méthode peut néanmoins être utilisée pour sérotyper les échantillons.Une analyse différentielle en ICPL (Isotope Coded Protein Labelling) des protéines plasmatiques de patients FC ou FS a permis d’identifier trois marqueurs potentiellement liés la sévérité de la maladie qui ont été validés en ELISA. La synthèse du virus mettant en jeu des partenaires cellulaires, le viroprotéome associé à chaque pathologie a été caractérisé par LC-MS/MS. Une méthode de purification du virus de la dengue a préalablement été mise au point sur un modèle in vitro. Cette méthode a ensuite été appliquée sur des pools de plasmas de patients prélevés en phase virémique. Des protéines virales ainsi que des protéines de l’hôte potentiellement associées aux virus ont été identifiées. Après analyse, une « empreinte » mettant en jeux des voies canoniques spécifiques a pu être déterminée pour les FS. Des ELISA ont été réalisés afin de valider le différentiel d’expression sur un choix de protéines.Au-delà de l’identification d’outils susceptibles d’aider les praticiens à poser un pronostic, ces études s’inscrivent dans la compréhension des mécanismes complexes qui sous-tendent le passage d’une FC à une FS. / Dengue is a endemic viral disease in tropical countries. If its classical form (CF) is benign, its severe form (SF) leads however to serious complications. Currently, the prognosis of severe dengue is unreliable. Differential proteomic studies on acute CF and FS plasma specimens were performed in order to identify early markers of progression to severe forms.The non-structural protein 1 (NS1) is a viral protein associated with pathogenicity. A method using SELDI-TOF/MS (Surface Enhanced Laser Desorption Ionization-Time of Flight/Mass Spectrometry) coupled to anti-NS1 monoclonal antibodies was developed in order to profile the proteins interacting with NS1 in plasma. Whereas NS1 protein was specifically detected in acute dengue plasma specimens, no NS1 ligand was identified. This method however allowed for sample serotyping.Plasma proteins of SF and CF patients were analyzed differentially using ICPL (Isotope Coded Protein Labeling). Three markers potentially related to disease severity were identified and validated by ELISA.As cellular partners are involved in virus biosynthesis, the viroproteome associated to each disease was characterized by LC-MS/MS. A method of dengue virus purification was first developed on an in vitro model. This method was then applied to pools of acute plasma of either CF or SF patients. We identified viral proteins as well as host proteins potentially associated with the viral particles. A footprint involving specific canonical pathways were subsequently identified in SF patients. Finally, a set of proteins found differentially expressed was validate by ELISA.Beyond identification of tools allowing assessment of dengue severity prognosis, these results give clues on the complex mechanisms underlying the transition from the CF to the SF.

No description available.

Research on sexual behavior in late life is limited but is growing. Despite ageist stereotypes associating old age with asexuality, older adults continue to desire and engage in sexual behavior. Previous studies have examined the relationship between health and the ability to engage in sexual behavior, sexual satisfaction, desire, or interest in sex. Research has yet to examine the potential reverse of this relationship, where sexual behavior may serve as a protective factor against health outcomes. This dissertation examined three research questions to test the relationship between sexual activity or hugging and self-reported health outcomes (e.g., arthritis and diabetes) and biomarkers (e.g., C-reactive protein (CRP) and hemoglobin (HbA1c)). The first question explored whether sexual activity or hugging is associated health outcomes. The second question explored whether social support modifies the relationship between sexual activity or hugging and health outcomes. The final question explored whether sexual activity or hugging and demographic or health variables interact as moderators to health outcomes. This study examined data of older adults between 57 and 85 years, from two waves of the National Social Life, Health, and Aging Project (NSHAP). The subjective health outcomes were self-reported arthritis and self-reported diabetes diagnoses. Objective health measures were analyzed using biomarkers. Both C-reactive protein and HbA1c were collected from dried blood spots. The main independent variables of interest were sexual activity and hugging. Sexual activity was assessed by combining participant responses to frequency of intercourse, foreplay and masturbation in the last 12 months. Hugging was assessed by participant responses to frequency of close physical contact over the last 12 months. Results from question one indicated at wave 1, engaging in hugging was associated with higher likelihood of self-reported arthritis (OR = 1.23, p = 0.029), while sexual activity was associated with decreased likelihood of diabetes (OR = 0.61, p < 0.001). Examining objective markers of health, sexual activity was associated with 0.25 points lower HbA1c level (p < 0.001). Although some significant results were found at wave 1, none of the associations remained significant when examining change in health at wave 2. The findings from question two examined the interaction of social support and sexual activity or hugging on health outcomes. The interactions between social support and sexual activity or social support and hugging did not significantly influence health at wave 1 or change in health at 2. Question three examined potential moderators of the relationship between sexual activity or hugging and health, including age, race, education, income, gender, and hours of sleep. At wave 1, the findings showed a significant interaction between race and sexual activity associated with a decreased likelihood of reporting arthritis (OR = 0.79, p = 0.021). A significant interaction of age and sexual activity was associated with an increased likelihood of diabetes (OR = 1.04, p = 0.008). At wave 2, a significant interaction between age and hugging was associated with decreased likelihood of arthritis (OR = 0.97, p = 0.006), while the interaction term of sleep and hugging was associated with increased likelihood of diabetes (OR = 1.19, p = 0.015). The results from this study show some support for the exploration of a potential bi-directional pathway between sexual activity or hugging and health. This study provides some evidence indicating the importance of understanding the role of sexual activity or hugging in the lives of older adults, and the possible influence on physical health. Future studies should continue exploring this pathway indicating possible benefits of engaging in sexual activity or hugging on health, and a larger impact on quality of life for older adults who desire to maintain intimate relationships in late life.

The study of circulating biomarkers and their association with disease outcomes has become progressively complex due to advances in the measurement of these biomarkers through multiplex technologies. Although the availability of numerous serum biomarkers is highly promising, multiplex assays present statistical challenges due to the high dimensionality of these data. In this dissertation, three studies are presented that address these challenges using L1 penalized regression methods. In the first part of the dissertation, an extensive simulation study is performed for the logistic regression model that compares the Least Absolute Shrinkage and Selection Operator (LASSO) method with five LASSO-type methods given scenarios that are present in serum biomarker research, such as high correlation between biomarkers, weak associations with the outcome, and sparse number of true signals. Results show that choice of optimal LASSO-type method is dependent on data structure and should be guided by the research objective. Methods are then applied to the Tucson Epidemiological Study of Airway Obstructive Disease (TESAOD) study for the identification of serum biomarkers of overweight and obesity. Measurement of serum biomarkers using multiplex technologies may be more variable as compared to traditional single biomarker methods. Measurement error may induce bias in parameter estimation and complicate the variable selection process. In the second part of the dissertation, an existing measurement error correction method for penalized linear regression with L1 penalty has been adapted to accommodate validation data on a randomly selected subset of the study sample. A simulation study and analysis of TESAOD data demonstrate that the proposed approach improves variable selection and reduces bias in parameter estimation for validation data as small as 10 percent of the study sample. In the third part of the dissertation, a measurement error correction method that utilizes validation data is proposed for the penalized logistic regression model with the L1 penalty. A simulation study and analysis of TESAOD data are used to evaluate the proposed method. Results show an improvement in variable selection.

Doctor of Philosophy / Department of Animal Sciences and Industry / Jim L. Nelssen / Four experiments using 350 pigs were conducted to determine the effects of dietary nutrients on the incidence of osteochondrosis (OC) and to evaluate the use of biomarkers to predict its occurrence in growing-finishing pigs. The first experiment was conducted to evaluate the potential of dietary ingredients with known functions in cartilage and bone metabolism on incidence of OC in pigs (PIC 327 × 1050, initially 39 kg). Results suggest that pigs fed high levels of added copper and manganese, silicon, methionine and threonine, or proline and glycine had reduced OC severity scores. A second experiment evaluated other dietary ingredients that may impact OC as well as with or without ractopamine HCl (PIC 327 × 1050, initially 47 kg). Results suggest that feeding a combination of added methionine, manganese, proline, and glycine or added silicon can numerically reduce OC severity scores compared to pigs fed a standard corn-soybean meal based diet. Feeding ractopamine HCl did not affect the incidence or severity of OC. A third experiment was conducted to evaluate the effect of dietary lysine concentration with or without the addition of high methionine, manganese, and copper in a 3 × 2 factorial arrangement in growing-finishing pigs (PIC 327 × 1050, initially 41 kg). Results suggest that increasing dietary lysine concentrations increased the severity of OC. Furthermore, feeding additional methionine, manganese, and copper reduced OC severity scores. A fourth experiment was conducted to determine the usefulness of ten different biomarkers of cartilage and bone metabolism to predict the incidence of OC and the correlation of individual markers with the severity of OC. Results suggest that measuring serum C-propeptide of type II collagen (CPII) will predict animals with OC, as gilts with a two-fold increase in CPII are 97 times more likely to have OC. As well, serum collagen type II carboxy-terminal 3/4 long fragment (C2C) explained 49% of the variation in OC severity scores. The results of these experiments suggest that feeding added copper, manganese, methionine, silicon, or proline and glycine may reduce OC severity scores in pigs, while measuring biomarkers CPII and C2C can aid in determining animals with OC.

Philosophiae Doctor - PhD / Breast cancer is one of the major causes of death in South Africa. About 1 in 29 South African women are at risk of developing this type of cancer in their lifetime. The global incidence of breast cancer also increases annually with over 1 million new cases diagnosed every year. Molecular diagnostic techniques such as qRT-PCR, Fluorescent In Situ Hybridization (FISH), Immunohistochemistry (IHC) and ELISA are used to diagnose breast cancer. Some of these diagnostic techniques make use organic fluorophores as fluorescent reporter molecules. The principle of all these diagnostic techniques is reliant on the detection of molecular biomarkers that are associated with the disease. In most cases these molecular biomarkers are DNA, RNA or proteins that are up-regulated in response to or as a result of the disease. The first aim of this study was therefore to identify membrane proteins that are up-regulated in cancers that can potentially be used as biomarkers for the detection of breast cancer. The second aim of this study was to investigate the application of quantum dots in the development of a molecular diagnostic test that can detect a breast cancer biomarker. The most commonly used method to identify molecular biomarkers for diseases have traditionally been gene expression analysis using technologies such as DNA microarray. These technologies have certain limitations and have therefore not been very successful in identifying useful disease biomarkers. Biomarker II discovery by proteomics can overcome some of these limitations and is potentially a more suitable method to identify molecular biomarkers for breast cancer. In this study proteomics in combination with Stable Isotope Labelling with Amino Acids in Cell Culture SILAC was used to do a comparative analysis of the expression levels of membrane proteins present in a human breast cancer cell line (MCF-7) derived from a breast cancer patient and a human breast cell line (MCF- 12A) derived from a healthy individual. This led to the identification of the transmembrane protein, GFRA1 as potential new biomarker for breast cancer. This study showed that this protein is over expressed in MCF-7 cells as compared to MCF-12A cells and that it is also highly expressed in the myoepthelial cells of the milk ducts of breast cancer patients. This study also demonstrates the use of molecular beacon technology to develop a DNA probe for the detection of cDNA encoding the CK19 gene, which is a known biomarker for breast cancer. In the development of this probe, quantum dots were used as the fluorescence reporter. This molecular beacon probe was able to demonstrate the over expression of CK19 in MCF-7 cells. This study shows that this technology can potentially be used as a diagnostic test for breast cancer and since quantum dots are used in the development of these molecular beacon probes, this diagnostic test can potentially facilitate the development of multiplex detection systems for the diagnosis of breast cancer. Molecular beacon technology can potentially also be used to detect novel biomarkers such as GFRA1.