The effect of stage of lactation on milk yield, somatic cell counts, mineral and fatty acid profiles in pasture-based Friesian, Jersey and Friesian × Jersey cows was investigated. Twenty Friesian, twenty Jersey and twenty Friesian × Jersey cows were randomly selected from a dairy herd. A total of 202 milk samples were collected and analysed in three stages of lactation. Genotypic differences were observed in milk yield and fat content. Friesian cows produced the highest yield and lowest fat content whereas the opposite was true for Jersey cows (P<0.01). No significant differences were observed in SCC in the different genotypes, but SCC levels were higher in mid and late lactation (P<0.001). There was no effect (P<0.005) of genotype and stage of lactation interaction on Ca, P, Mg, Na, Mn and Bo concentration. Jersey cows had the least concentration of Fe and Cu in all stages of lactation. Strong positive correlations were observed among Ca and P, Mg and Zn. Aluminium had a strong positive relationship with Bo, Fe, Mn and Zn (P<0.001). Generally, SCC had a weak positive relationship with macro elements but a significant negative relationship with microelements. Yield levels were negatively correlated with Mg, Na, Al, Mn and SCC. Pasture ALA, SFA, n-3, n-6/n-3 and PUFA/MUFA concentration did not differ across the study period. Linoleic acid was highest in the second phase which coincides with mid lactation in cows (P<0.001). Highest moisture content coincided with the least fat free dry matter content in early lactation (P<0.001). Significantly high fat content was observed in late lactation than in early lactation. Highest butyric, caproic, linoleic, n-6 and PUFA were observed for Friesian cows. All other fatty acids ratios were not significantly different among different genotypes. Highest CLA, ALA, LA, SFA, PUFA, n-6, and n-3 and atherogenicity index were observed in early lactation whereas desaturase activity indices were highest in late lactation. Strong positive correlations were observed among milk vaccenic, ALA, LA and CLA concentrations. Inverse relationships were observed between SFA and long chain fatty acids. It can be concluded, it may be of advantage to consume milk from early stage of lactation poses a lower risk to coronary diseases and are much safer to consume.

Orientador: Mary Ann Foglio / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba / Made available in DSpace on 2018-08-26T10:27:31Z (GMT). No. of bitstreams: 1 Lloret_FelipeCyrillo_M.pdf: 3758612 bytes, checksum: 7002aac1760fd55833ce91a085d63599 (MD5) Previous issue date: 2014 / Resumo: A Pterodon pubescens Benth, conhecida como sucupira é nativa das regiões que consiste o cerrado brasileiro, nos estados de Minas Gerais, São Paulo, Goiás e Mato Grosso do Sul. Nosso grupo iniciou estudos com a espécie Pterodon pubescens Benth em 1998. Inicialmente foram desenvolvidos estudos relacionado a eficácia dos seus extratos e determinação dos princípios ativos. O objetivo deste trabalho foi avaliar o efeito do calor sobre o processo de formação e degradação dos componentes vouacapanos nos frutos de Pterodon pubescens Benth. e sua influência sobre a atividade antiproliferativa in vitro em células tumorais humanas. Os frutos de P. pubescens foram coletados no município de São Carlos, localizado no estado de São Paulo, e no município de Ponto Chique, localizado no estado de Minas Gerais no período de agosto e setembro de 2012. Mensalmente foram preparados extratos por sistema Soxhlet e maceração dinâmica com as amostras de frutos provenientes dos municípios de São Carlos e de Ponto Chique armazenados em estufa, analisados por cromatografia gasosa capilar acoplada a um detector seletivo de massas e tiveram suas atividades antiproliferativas contra linhagem tumoral humana (PC-3) avaliadas por método de sulforrodamina B. Todos os resultados de teores dos compostos de interesse obtidos por CG-EM foram submetidos a análise de variância de uma única via (ANOVA), considerando-se como nível crítico p? 0,05 para que seja considerados diferença significante entre os grupos estudados, seguidos do Teste de Duncan. Para a análise dos compostos que possuem uma maior influência na atividade antiproliferativa do nos extratos obtidos foi utilizado a Análise de Componentes Principais. Foi possível concluir que há uma possível variação genética das espécies de Pterodon pubescens Benth., e que o calor não influenciou na formação do 6?- hidróxi- 7?- acetoxivouacapano, porém influencia no teor dos outros compostos estudados / Abstract: Pterodon pubescens Benth (Fabeacea) chemical variability was investigated to understand if the secondary metabolites 6?-acetoxi 7?-hydroxy-vouacapan content variability, was produced as an artefact overtime or was characteristic of genotype variability. The oil was extracted both by soxhlet system and at room temperature with stirring. The chemical composition was monitored by gas chromatography-mass spectrometry. The samples were tested in an anticancer assay against prostate (PC-03) human cancer cell line. The response parameter (TGI) was calculated. The chemical variability data was evaluate using the statistical analysis using Principal Component Analysis (PCA) to understand which of the extract's components had a real impact on the in vitro antiproliferative activity. Hierarchical Cluster Analysis (HCA) was performed to visualize the similarities among the extracts components with TGI values. Unscrumbler® v. 9.7 software was employed for analysis. The chemical composition of samples from São Carlos, São Paulo and Ponto Chique, Minais Gerais were monitored monthly during one year for Geranilgeraniol, 6?-acetoxi 7?-hydroxy-vouacapan, 6?- hydroxy-7?-acetoxy-vouacapan-17?-oate methyl ester and 6?-acetoxy-7?-hydroxyvouacapan- 17?-oate methyl ester. In vitro cytotoxicity screening against human prostate cancer cell line displayed higher selectivity and potent anticancer activity with TGI 11.43 mg ml?1 when higher 6?-acetoxi 7?-hydroxy-Vouacapan over total Voaucapan ratio (3.14) was achieved. Nevertheless 6?-acetoxi 7?-hydroxyvouacapan maintained approximately the same content throughout the year among the samples in opposition to overall Voaucapan content. The total Vouacapan total content was directly proportional to Geranygeraniol content decrease content. Samples from São Carlos at time zero had 26% geranylgeraniol content whereas Minas Gerais samples contained the highest content of 1.3%. Throughout the stability test geranylgeraniol concentration decreased with a straight relationship of overall increase of vouacapan content. This study highlights the complexity of factors involved in the production of secondary metabolites in plants. To enable the development of an herbal medicine, one of the greatest challenges is plant input standardization in order to meet efficacy, safety and reproducibility final product's requirements as recommended by Brazilian Sanitary Regulatory Agency (ANVISA). Data presented herein suggest that 6?-acetoxi 7?-hydroxy-vouacapan content variability has a straight relationship with genotype other than produced as artefact overtime / Mestrado / Farmacologia, Anestesiologia e Terapeutica / Mestre em Odontologia

A partir da finalização do projeto genoma humano, fornecendo informação sobre o código genético de um indivíduo referência, pesquisadores direcionam a busca por sítios polimórficos existentes no DNA genômicos e mitocondrial capazes de influenciar direta ou indiretamente traços relevantes nas mais diversas áreas. No esporte esta busca tem sido direcionada para identificar marcadores genéticos associados aos fenótipos envolvidos com o desempenho físico (endurance vs. força e potência muscular). Atualmente, diversos polimorfismos foram associados ao desempenho físico-esportivo, em pelo menos um estudo. No entanto, como o rendimento desportivo é um fenômeno poligênico, ou seja, sob a regulação de vários genes, a associação de um polimorfismo é insuficiente para caracterizar o atleta. Hipóteses estão sendo desenvolvidas na tentativa de identificar o perfil poligênico (combinação de vários polimorfismos) em grupos de atletas. Alguns pesquisadores avaliaram o perfil poligênico preliminar de atletas europeus, analisando um conjunto de 7 a 10 polimorfismos, entretanto, esses achados precisam ser investigados em outras populações. O objetivo deste projeto foi traçar o perfil poligênico de atletas brasileiros. Para tanto, uma coorte brasileira envolvendo 342 atletas de endurance, 308 atletas de força e potência, 93 atletas de esportes coletivos, 165 atletas de lutas e 967 não atletas foi considerada neste estudo. O DNA genômico destes indivíduos foram extraídos e 34 polimorfismos foram genotipados. De uma forma geral, o grupo Endurance foi bem semelhante ao grupo Não atletas. Foi verificado que os polimorfismos AGT M235T - Alelo G, ACTN3 R577X - Alelo R, MCT1 D490E - Alelo T, PPARGC1A G482S - Alelo C, VEGFR2 Q472H - Alelo T, CNDP2 C/G - Alelo G e PPARA G/C - Alelo C podem ser considerados marcadores genéticos para atletas de atividades que envolvam a força e a potência muscular. Ainda, dos sete marcadores genéticos associados ao grupo Força/Potência, dois deles foram associados em coortes estrangeiras ao grupo Endurance, sugerindo que a população Brasileira pode apresentar associações específicas e diferenciadas. O escore total dos genótipos (TGS; modelo aditivo), utilizando os sete polimorfismos mais relevantes para a força e potência, foi eficiente em discriminar o grupo alvo, dos demais grupos. Foi observado que possuir uma quantidade >= 9 alelos associados (escore >= 64,3) aumentou significativamente a chance (76,8%) do indivíduo pertencer ao grupo Força/Potência. No entanto, existem atletas do grupo alvo, de destaque internacional (top atletas), que possuem um baixo número de alelos associados (2 a 4 alelos associados), bem como existem atletas de endurance que possuem um alto número de alelos associados (10 a 12 alelos associados). Possuir um número maior de alelos associados ao fenótipo alvo pode ser uma vantagem para o atleta, que precisa ser adequadamente estimulado por fatores ambientais. Contudo, uma associação genótipo-fenótipo não indica que o indivíduo pode antecipar uma carreira vitoriosa. O desempenho físico-esportivo reflete uma complexa interação de fatores ambientais ou sociais / After the conclusion of the human genome project, information about the genetic code of an individual (reference) was provided. Since then, scientists directed their research to identify polymorphic sites in genomic and mitochondrial DNA able to be associated with relevant phenotypes. In sports, the search has been directed to identify genetic markers associated with performance-related phenotypes (endurance vs. muscle strength and power phenotypes). Currently, many polymorphisms have been associated with physical performance in at least one study. However, as the sport performance is a polygenic phenomenon (i.e., under the regulation of several genes), the association of a single polymorphism is insufficient to characterize the athlete biology. Hypotheses are being developed in an attempt to identify the athlete polygenic profile (i.e., the combination of several sports-relevant polymorphisms). Some researchers evaluated a preliminary polygenic profile of European athletes by analyzing a range from 7 to 10 polymorphisms; however, these findings need to be investigated in other populations. The objective of this project was to explore the polygenic profile of Brazilian athletes. A Brazilian cohort involving 342 endurance athletes, 308 strength and power athletes, 93 athletes from team sports, 165 combat sports athletes and 967 non-athletes was evaluated. Genomic DNA of these individuals was extracted and 34 polymorphisms were genotyped. In general, the Endurance group was very similar to the Non-athlete group. The main associations were found when comparing the Strength/Power group with the Non-athlete group. Seven genetic markers were highlighted in the Strength/Power group. It was found that the following polymorphisms AGT M235T - Allele G, ACTN3 R577X - Allele R, MCT1 D490E - Allele T, PPARGC1A G482S - Allele C, VEGFR2 Q472H - Allele T, CNDP2 C/G - Allele G e PPARA G/C - Allele C can be considered genetic markers for Brazilian strength and power athletes. Of the seven genetic markers associated with Strength/Power, two were previously associated in other cohorts to Endurance, suggesting that the Brazilian population may have different and specific associations. The Total Genotype Score (TGS; additive model) using the seven most relevant polymorphisms was efficient to discriminate the Strength/Power group. It was observed that having a quantity >= 9 alleles associated (score >= 64.3) significantly increased the chance (76.8%) of belong to the Strength/Power group. However, there are some athletes of the Strength/Power group (international-level) showing a low number of associated alleles (2 to 4 associated alleles), as well as Endurance athletes showing a high number of associated alleles (10 to 12 associated alleles). A high number of associated alleles may be advantageous to the athlete; however they need to be properly stimulated by environmental factors. Any genotype-phenotype association does not indicate that an individual can anticipate a successful career. The athletic performance reflects a complex interaction of genetic and environmental / social factors

As conexinas (Cx) compõem as junções comunicantes do tipo gap, entre elas a Cx43 é a mais prevalente. A diminuição de sua expressão está relacionada com diversas alterações fisiológicas. Sua importância in vivo foi relatada em camundongos com deleção de um dos alelos de Cx43 (Cx43+/-), pois os animais Cx43-/- não são viáveis. Devido esta inviabilidade técnica, os estudos com essa proteína foram realizados com animais Cx43+/-. Assim, este trabalho propõe a criação de um modelo transgênico para o estudo da Cx43. Para isso, o gene Cx43 foi reintroduzido ao genoma murino, porém regulado pelo sistema induzido por doxiciclina TetOn. Vetores de expressão gênica foram construídos e validados quanto sua funcionalidade in vitro e posterior transferencia para zigotos murinos por meio de microinjeção pronuclear. Obtivemos sucesso na construção do sistema de expressão. A funcionalidade dos vetores foi confirmada in vitro utilizando células HeLa e E10. Nos experimentos in vivo, apesar das taxas adequadas de nascimento nenhum deles apresentou genótipo positivo para o transgene. / The connexins (Cx) comprise gap junctions type, and the Cx43 is the most prevalent. The decrease of its expression is related to various physiological changes. Its importance in vivo has been reported in mice with deletion of one allele of Cx43 (Cx43+/-), because the animals Cx43-/- are not viable. So all studies with this protein were performed with animals Cx43+/-. Thus, this paper proposes the creation of new a transgenic model for the study of Cx43 protein. For this, the Cx43 gene was reintegrated to the murine genome, but drived by doxycycline Teton inducible system. Gene expression vectors were constructed and validated in vitro and subsequent transfer to mouse zygotes by pronuclear microinjection. We got success in vector construction and functionality. The functionality of the vectors was confirmed in vitro using HeLa cells and E10. In experiments in vivo, despite adequate rates of birth, no one animal showed positive genotype for the transgene.

O experimento consistiu de dezesseis tratamentos distribuídos num esquema fatorial 4x2x2, sendo 4 genótipos (Excel (E) - Nn, Linha 21 (L21) - nn, Maximus (M) - NN e Optimus (O) - NN), 2 sexos (machos castrados e fêmeas) e 2 pesos (leves – 95 a 100Kg e pesados – 115 a 120Kg) e seis repetições. Foram realizadas medidas de composição da carcaça: Peso da Carcaça Resfriada (PCR), Área de Olho de Lombo (AOL), Comprimento de Olho de Lombo (COL), Profundidade de Toucinho (PT), Porcentagem de Carne Magra (%CM) e medidas de Espessuras de Músculo (EM) e Gordura (EG) e avaliações de qualidade de carne: pH, cor (L*a*b*), Perda por Exsudação (PE) e Capacidade de Retenção de Água (CRA). Os resultados estatísticos da composição da carcaça para o genótipo nn indicaram valor de AOL significativamente superior em relação aos genótipos Nn, NNM e NNO e valor de PT significativamente menor que NNM e NNO, porém, não diferindo de Nn. Quanto à EM, ocorreu uma tendência do NNM apresentar os maiores valores, seguido pelo Nn, nn e NNO. Em relação à EG, NNO mostrou os maiores valores, não diferindo significativamente de NNM e nn, enquanto o genótipo Nn apresentou o menor valor para esta característica. A %CM dos genótipos nn e Nn foi significativamente maior que NNM e NNO, confirmando a expressão do gene recessivo (n) em depositar mais carne na carcaça que o dominante (N). Os resultados estatísticos de qualidade de carne mostraram que o genótipo nn apresentou o menor valor de pH24 no músculo Semimembranosus, combinado com os maiores valores de luminosidade (L*) e PE em comparação com Nn, NNM e NNO. Não houve diferença significativa entre os genótipos NN and Nn na PE. Em relação à CRA, NNM obteve valor significativamente menor que Nn, nn e NNO. Os resultados obtidos permitem concluir que o gene halotano em homozigose (L21) contribuiu para a obtenção de uma carcaça mais musculosa, porém, com qualidade de carne inferior, apresentando maior incidência de carne PSE. Em relação ao sexo, os machos resultaram carcaças mais pesadas, mais gordas e carne com menor luminosidade. Finalmente, com exceção da %CM, os valores de composição da carcaça aumentaram com o peso de abate, não sendo constatadas mudanças nas características de qualidade avaliadas. / This experiment consisted of sixteen treatments in a 4x2x2 factorial arrangement, 4 genotypes (Excel – Nn, Line 21 – nn, Maximus – NN and Optimus – NN), 2 sexes (barrows and gilts) and 2 weights (light – 95-100Kg and heavy – 115-120Kg) and 6 repetition. The exams were submitted to the following carcass composition measurements: Cold Carcass Weight (CCW), Loin Eye Area (LEA), Loin Eye Length (LEL), Fat Depth (FD), Lean Meat Percentage (LM%) and Fat (FT) and Muscle (MT) Thickness. Meat quality measurements were also realized: pH, Color (L*a*b*), Drip Loss (DL) and Water Holding Capacity (WHC). The results of this study showed that genotype nn had shown significantly higher values of LEA than another genotypes and significantly lower values of FD than NNM and NNO, and it didn’t differ from genotype Nn. There was a tendency by genotype NNM to show higher values of MT followed by Nn, nn and NNO. Genotype NNO showed higher values to FT and did not differed significantly from NNM and nn, while genotype Nn showed lower values to this characteristic. Genotypes nn and Nn showed significantly higher values of LM% than NNM and NNO. With reference to meat quality, genotype nn has shown lower value of pH24 measured in the muscle Semimenbranosus and higher values of L* and DL than genotypes Nn, NNM and NNO. Genotype NNM had significantly higher values of WHC than nn, Nn and NNO. There was no significant difference between NN and Nn when considered DL. The results above indicate that halothane gene in homozygous (nn) helped to obtain more muscular carcasses. On the other hand, this genotype showed worst meat quality. As a consequence of that, genotype nn has shown higher incidence of pale, soft and exudative meat. With reference to sex, barrows had significantly higher values to CCW, FD, FT, L* and b*, resulting paler meat and heavier and fatter carcass than gilts. Except for LM%, as carcass weight increased there were increases in all the characteristics. The data from this study suggest that there were no meat quality differences between the two slaughter weights.

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第22002号 / 医博第4516号 / 新制||医||1038(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 滝田 順子, 教授 松田 文彦, 教授 山田 亮 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

MSCAGR (Animal Science) / Department of Animal Science / See the attached abstract below

A variety of modern software packages are available for genotype imputation relying on advanced concepts such as pre-phasing of the target dataset or utilization of admixed reference panels. In this study, we performed a comprehensive evaluation of the accuracy of modern imputation methods on the basis of the publicly available POPRES samples. Good quality genotypes were masked and re-imputed by different imputation frameworks: namely MaCH, IMPUTE2, MaCH-Minimac, SHAPEIT-IMPUTE2 and MaCH-Admix. Results were compared to evaluate the relative merit of pre-phasing and the usage of admixed references. We showed that the pre-phasing framework SHAPEIT-IMPUTE2 can overestimate the certainty of genotype distributions resulting in the lowest percentage of correctly imputed genotypes in our case. MaCH-Minimac performed better than SHAPEIT-IMPUTE2. Pre-phasing always reduced imputation accuracy. IMPUTE2 and MaCH-Admix, both relying on admixed-reference panels, showed comparable results. MaCH showed superior results if well-matched references were available (Nei’s GST ≤ 0.010). For small to medium datasets, frameworks using genetically closest reference panel are recommended if the genetic distance between target and reference data set is small. Our results are valid for small to medium data sets. As shown on a larger data set of population based German samples, the disadvantage of pre-phasing decreases for larger sample sizes.

The concept of a fitness landscape arose in theoretical biology, while that of effective fitness has its origin in evolutionary computation. Both have emerged as useful conceptual tools with which to understand the dynamics of evolutionary processes, especially in the presence of complex genotype-phenotype relations. In this contribution we attempt to provide a unified discussion of these two approaches, discussing both their advantages and disadvantages in the context of some simple models. We also discuss how fitness and effective fitness change under various transformations of the configuration space of the underlying genetic model, concentrating on coarse-graining transformations and on a particular coordinate transformation that provides an appropriate basis for illuminating the structure and consequences of recombination.

Finding a relation between a particular phenotype and genotype is one of the central themes in medical genetics. Single-nucleotide polymorphisms are easily assessable markers allowing genome wide association (GWA) studies and meta-analysis. Hundreds of such analyses were performed in the last decades. Even though several tools for such analyses are available, an efficient SNP-data transformation tool was tool was necessary. We developed a data management tool fcGENE which allows us easy transformation of genetic data into different formats required by different GWA tools. Genotype imputation which is a common technique in GWA, allows us to study the relationship of a phenotype at markers that are missing and even at completely un-typed markers. Moreover this technique helps us to infer both common and rare variants that are not directly typed. We studied different aspects of the imputation processes especially focussing on its accuracy. More specifically, our focus lied on the impact of pre-imputation filtering on the accuracy of imputation results. To measure the imputation accuracy, we defined two new statistical sores, which allowed us the comparison between imputed and true genotypes directly. Our direct comparison between the true and imputed genotypes showed that strict quality filtering of SNPs prior to imputation process may be detrimental. We further studied the impact of differently selected reference panels from publicly available projects like HapMap and 1000 genome projects on the imputation quality. More specifically, we analysed the relationship between genetic distance of the reference and the resulting imputation quality. For this purpose, we considered different summary statistics of population differentiation (e.g. Reich’s , Nei’s and other modified scores) between the study data set and the reference panel used in imputation processes. In the third analysis, we compared two basic trends of using reference panels in imputation process: (1) use of genetically best-matched reference panel, and (2) use of an admixed reference panel that allows the use of individual reference panel from all possible type of populations, and let the software itself select the optimal references in a piece-wise manner or as complete sequences of SNPs for each individual separately. We have analysed in detail the performance of different imputation software and also the accuracy of the imputation processes in both cases. We found that the current trend of using software with admixed reference panel in all cases is not always the best strategy. Prior to imputation process, phasing of study data sets by using an external reference panel is also a common trend especially when it comes to the imputation of large datasets. We studied the performance of different imputation frameworks with or without pre-phasing. It turned out that pre-phasing clearly reduces the imputation quality for medium-sized data sets.:Table of Contents List of Tables IV List of Figures V 1 Overview of the Thesis 1 1.1 Abstract 1 1.2 Outlines 4 2 Introduction 5 2.1 Basics of genetics 5 2.1.1 Phenotype, genotype and haplotype 5 2.1.2 Hardy-Weinberg law 6 2.1.3 Linkage disequilibrium 6 2.1.4 Genome-wide association analysis 7 2.2 Phasing of Genotypes 7 2.3 Genotype imputation 8 2.3.1 Tools for Imputing genotype data 9 2.3.2 Reference panels 9 3 Results 11 3.1 Detailed Abstracts 11 3.1.1 First Research Paper 11 3.1.2 Second Research Paper 14 3.1.3 Third Research Paper 17 3.1.4 Fourth Research Paper 19 3.2 Discussion and Conclusion 22 4 Published Articles 27 4.1 First Research Paper 27 4.1.1 Supplementary Information 34 4.2 Second Research Paper 51 4.2.1 Supplementary Information 62 4.3 Third Research Paper 69 4.3.1 Supplementary Information 85 4.4 Fourth Research Paper 97 4.4.1 Supplementary Information 109 5 Zusammenfassung der Arbeit 117 6 Bibliography 120 7 Eigene Publikationen 124 8 Darstellung des eigenen Beitrags 125 8.1 First Research Paper 125 8.2 Second Research Paper 126 8.3 Third Research Paper 127 8.4 Fourth Research Paper 128 9 Erklärung über die eigenständige Abfassung der Arbeit 129 10 Danksagung 130 11 Curriculum Vitae 131 List of Tables IV List of Figures V 1 Overview of the Thesis 1 1.1 Abstract 1 1.2 Outlines 4 2 Introduction 5 2.1 Basics of genetics 5 2.1.1 Phenotype, genotype and haplotype 5 2.1.2 Hardy-Weinberg law 6 2.1.3 Linkage disequilibrium 6 2.1.4 Genome-wide association analysis 7 2.2 Phasing of Genotypes 7 2.3 Genotype imputation 8 2.3.1 Tools for Imputing genotype data 8 2.3.2 Reference panels 8 3 Results 8 3.1 Detailed Abstracts 8 3.1.1 First Research Paper 8 3.1.2 Second Research Paper 8 3.1.3 Third Research Paper 8 3.1.4 Fourth Research Paper 8 3.2 Discussion and Conclusion 8 4 Published Articles 8 4.1 First Research Paper 8 4.1.1 Supplementary Information 8 4.2 Second Research Paper 8 4.2.1 Supplementary Information 8 4.3 Third Research Paper 8 4.3.1 Supplementary Information 8 4.4 Fourth Research Paper 8 4.4.1 Supplementary Information 8 5 Zusammenfassung der Arbeit 8 6 Bibliography 8 7 Eigene Publikationen 8 8 Erklärung über die eigenständige Abfassung der Arbeit 8 9 Danksagung 8 10 Curriculum Vitae 8