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Metal-organic frameworks (MOFs) have attracted much attention in the past few decades due to their ordered, crystalline nature, synthetic tunability, and porosity. MOFs represent a class of hybrid inorganic-organic materials that have been investigated for their applications in areas such as gas sorption and separation, catalysis, drug delivery, and electron or proton conduction. It has been the goal of my graduate research to investigate MOFs for their ability to transport electrons and store and separate gases for ultimate catalytic applications in alternative energy generation. I aim to provide new insight into the design and development of stable MOFs for such applications. We first investigated a cobalt(III) porphyrin based MOF comprised of Co(II)-carboxylate nodes, designated as CoPIZA, for its electron transport capabilities. Thin films of CoPIZA were formed solvothermally on conductive fluorine-doped tin oxide (FTO) substrates and used for electrochemical characterization. Electrochemistry coupled with spectroscopic analysis of the CoPIZA film revealed reversible reduction of the cobalt centers of the porphyrin linkers with maintenance of the overall framework structure. The mechanism of charge transport throughout the film was facilitated by redox hopping of electrons between the metal centers of the nodes and linkers. The ability to incorporate desired properties, such as pore functionalities or open metal centers, into frameworks makes them attractive for applications in separation of gaseous mixtures, such as CO2/N2 from combustion power plants. To investigate the selective adsorption properties, we performed gas sorption measurements on bulk MOF materials to determine their affinity toward CO2. Two Zn-based MOFs containing 2,5-pyridine dicarboxylate linkers were prepared in our laboratory and contained unsaturated Zn(II) metal centers, which possess a binding site on the metal without an activation procedure to remove bound solvent molecules. These MOFs were compared to the well-known Zn-based MOF-69C containing 1,4-benzene dicarboxylate linkers. Thermodynamic analysis of the gas sorption data revealed that the mechanism of CO2 binding involved the coordinatively unsaturated Zn(II) center. The microporous MOF also demonstrated selectivity for CO2 over N2 under the same conditions. As these materials were able to uptake CO2, their ability to transport electrons was also investigated for ultimate applications in catalysis. Electrochemical impedance spectroscopy was performed on the bulk MOF powders and was coupled with solid-state nuclear magnetic resonance spectroscopy. These results determined that the conduction mechanism proceeded via solvent molecules within the pores of the framework. The catalytic ability toward water oxidation of two MOFs was investigated electrochemically. Initial studies focused on a cobalt-based MOF comprised of 2-pyrimidinolate (pymo) linkers, designated as Co(pymo)2, which was prepared on FTO via drop-casting and used for electrochemical experiments. At applied anodic potentials, the CoII centers of Co(pymo)2 became oxidized to form a Co-oxide species on the electrode surface, which was found to be the active catalysis for water oxidation. Further investigations utilized a notably more stable Zr-based MOF with nickel(II) porphyrin linkers, designated as PCN-224-Ni. PCN-224-Ni was prepared solvothermally on FTO and used directly for electrochemical water oxidation. The mechanism of water oxidation at PCN-224-Ni proceeds via oxidation of the porphyrin macrocycle followed by binding of water to the Ni(II) center. Cooperative proton transfer to the Zr-oxo node facilitated water oxidation with the eventual release of O2. Thorough characterization revealed that PCN-224-Ni retained its structural integrity over the course of electrochemical catalysis. These results have allowed us a deeper understanding of the mechanisms of electron transport and conduction throughout frameworks. Specifically, the incorporation of metalloporphyrin molecules with redox active metal centers coupled with the presence of redox active metal nodes resulted in redox hopping charge transport throughout the MOF. In addition, the presence of solvent molecules in the pores of the framework provided an extended network for charge transport. We have gained insight into the structure-function relationship of MOFs for applications in selective gas sorption, where an unsaturated metal center serves as the binding site for gas molecules. Finally, through selection of the components that comprise the framework, a stable metalloporphyrin MOF was found to be capable of electrochemically facilitating the water oxidation reaction. As a result, we have gained valuable insight into the properties of frameworks necessary for charge transport and stability, which will allow for further improvements in the smart design of MOFs for catalytic applications. / Ph. D.

This thesis presents an experimental study of the superconducting proximity effect in sub-micrometer sized ferromagnetic discs. Such discs belong to a class of mesoscopic ferromagnets intermediate between microscopic magnets with dimensions below about 10nm that behave as single giant spins and macroscopic structures that are larger than approximately 1 micrometer where domains are formed to minimise stray fields. The magnetic structure of mesoscopic magnets is strongly dependent on their geometric shape, allowing for purposeful engineering of magnetic structures using modern lithographic techniques. The ground magnetic state of mesoscopic ferromagnetic discs is the magnetic vortex where unusual time-asymmetric triplet superconductivity is predicted to exist and survive up to the non-magnetic coherence length, that is orders in magnitude larger than the ferromagnetic singlet coherence length. Magnetic Force Microscopy (MFM) was used to directly study the magnetic structure of the discs. To detect the proximity effect in the vortices, Andreev interferometers were used with normal parts replaced with mesoscopic ferromagnetic discs in the magnetic vortex state. The samples were fabricated using electron-beam lithography and a modified shadow evaporation technique developed within this project, allowing the whole structure to be made with highly precise alignment, without breaking vacuum and avoiding redundant ferromagnetic elements disturbing the magnetic vortices. Observations were made of superconducting phase periodic oscillations in the conductance of the Andreev interferometers. Such oscillations provide unambiguous evidence of phase coherent electron transport through the ferromagnetic vortex. Finally, further experiments are discussed that would provide a more detailed understanding of the long range proximity effect in SFS junctions.

This diploma thesis is focused on topics of and respiratory chain and their processing as educational animations for secondary schools. In the theoretical part of the diploma thesis, supporting terms are defined (visualisations, interdisciplinary relations, animations). Next, themes are included in educational field and thematic unit in RVP G. Two analyses of the topic of respiratory chain are mentioned and evaluated in this part. Firstly, there is an analysis focused on the most used chemical and biological textbooks. The second one is focused on available online animations. For research purpose of the diploma thesis quantitative method was used - a questionnaire survey. Results of this survey are listed in the practical part. Practical part presents educational materials specified for support of education topics of respiration and respiratory chain. Stress on illustrative nature and interdisciplinarity is laid in these materials. The principal materials are educational animations which were made in program Adobe Flash Professional CS6. Study text was also written to these animations. Created educational animations give a complex view on respiratory process. These materials can be used in biology and chemistry subjects or in scientific courses. Animations include two tests giving feedback to...

Espécies reativas de oxigênio (EROs) são normalmente e continuamente geradas em mitocôndrias, majoritariamente na cadeia de transporte de elétrons (CTE). Harman (1956, 1972 e 1992) teorizou que os radicais livres gerados nas mitocôndrias seriam a principal causa do envelhecimento. De fato, durante o envelhecimento é observado um desequilíbrio entre formação e remoção de EROs, que resulta em estresse redox. Essa condição favorece a formação de lesões oxidadas no DNA, acarretando em mutagênese ou morte celular. Diversos mecanismos moleculares cooperam para o reparo de DNA. Duas vias de reparo de DNA lidam com a maioria das lesões: o reparo por excisão de base (BER) e o reparo por excisão de nucleotídeos (NER). A via BER corrige pequenas modificações de bases que surgem de reações de desaminação, alquilação e oxidação. A via NER é mais versátil, reconhecendo lesões que distorcem a dupla hélice de DNA, como danos induzidos por luz UV e adutos volumos. Pacientes xeroderma pigmentoso (XP-A a XP-G) herdam mutações em um de sete genes que codificam proteínas envolvidas na via NER, ou em um gene que codifica uma polimerase translesão (XP-V). A doença é caracterizada por fotosensibilidade e incidência elevada de neoplasias cutâneas. A proteína XPC atua na etapa de reconhecimento da lesão de DNA na subvia de reparo global do genoma (GG-NER), e sua mutação dá origem aos sintomas clássicos de XP. Novas funções de XPC foram recentemente descritas: i) atuando como cofator na via BER auxiliando as DNA glicosilases OGG1, TDG e SMUG; ii) atuando como cofator transcricional de elementos responsivos a Oct4/Sox2, RXR e PPARα; e iii) na adaptação metabólica na transformação de queratinócitos. Então, propusemo-nos a investigar as relações entre XPC e a manutenção da integridade do DNA mitocondrial, a sensibilidade celular a estresse redox mitocondrial e possíveis alterações bioenergéticas e redox. Para tal, padronizamos um ensaio in vitro de cinética de incisão em DNA plasmidial a fim de investigarmos o possível papel de XPC no reparo de lesões oxidadas em mtDNA. Porém, nossos dados revelaram que XPC não se encontra em mitocôndrias. Apesar disso, células XP-C são mais sensíveis ao tratamento com azul de metileno (AM), antimicina A (AA) e rotenona (ROT), que geram estresse redox mitocondrial. A sensibilidade à AA foi completamente revertida em células corrigidas. Células XP-C apresentaram alterações quanto ao uso dos complexos mitocondriais, com diminuição da taxa de consumo de oxigênio (OCR) via complexo I e um aumento da OCR via complexo II, dependente da presença de XPC. Ademais, a linhagem XP-C apresentou um desequilíbrio redox mitocondrial com maior produção de EROs e menor atividade de GPx. O DNA mitocondrial de células XP-C apresentou níveis elevados de lesão e deleção, que no entanto não retornaram aos níveis encontrados em células selvagens na linhagem XP-C corrigida. Observamos uma acentuada diminuição da expressão de PPARGC1A, um importante regulador de biogênese mitocondrial. Contudo, não foi possível determinar o mecanismo de supressão da expressão de PPARGC1A. Por fim, identificamos que o tipo de mutação em XPC pode estar associado a expressão de PPARGC1A. Esse estudo abre novas possibilidade na investigação do papel de proteína XPC, à parte da instabilidade genômica, na adaptação metabólica e desequilíbrio redox em direção da progressão tumoral. / Mitochondria continuously produce reactive oxygen species (ROS), mainly at the electron transport chain. Harman (1956, 1972 e 1992) proposed that normal aging is driven by increased mitochondrially generated free radicals. Indeed, during the course of aging there is an increased imbalance between formation and removal of ROS, leading to redox stress. This condition favours the formation of oxidized DNA lesions, given rise to mutations and cell death. Several molecular mechanisms cooperates to repair the DNA. Two DNA repair pathways deal with the majority of lesions: base excision repair (BER) and nucleotide excision repair (NER). The BER pathway corrects small base modifications that arise from deamination, alkylation and oxidation reactions. The NER pathway is more versitile, recognizing helix-distorting lesions, such as UV-induced damage and bulky adducts. Xeroderma pigmentosum (XP-A to XP-G) patients inherit mutations in one of seven protein-coding genes involved in NER pathway, or in a gene coding a translesion DNA polymerase (XP-V). Photosensitivity and a thousand-fold increased in the risk of developing cutaneous neoplasms are the main clinical features of XP. XPC protein functions in the recognition step of global genome NER (GG-NER) sub-pathway, and mutations in this gene lead to classical XP symptoms. Recently, it has been described that XPC acts: i) as a cofactor in BER pathway through functional interaction with DNA glycosylases OGG1, TDG and SMUG1; ii) as coactivator in transcription at Oct4/Sox2, RXR and PPARα responsive elements; iii) in metabolic shift during keratinocytes transformation. Thus, we sought to investigate a possible role for XPC in the maintenance of mtDNA integrity, cellular sensitivity to mitochondrial redox stress and eventual bioenergetic and redox changes. For this purpose, we established an in vitro plasmid incision assay to investigate the possible role of XPC in the repair of oxidized lesions in mitochondrial DNA. However, our data revealed that XPC did not localized in mitochondria. Nonetheless, XP-C cells are more sensitive to methylene blue, antimycin A (AA) and rotenone treatment, which induce mitochondrial redox stress. The XP-C sensitivity to AA was completely reverted in XPC-corrected cells. XP-C cells presented altered usage of mitochondrial complexes, with decreased oxygen consumption rate (OCR) via complex I and increased OCR through complex II, an XPC-dependent phenomenon. Furthermore, the XP-C cell line showed mitochondrial redox imbalance with increased ROS production and decrease GPx activity. MtDNA from XP-C cells accumulate lesions and deletions, which, however, were found at similar levels in the corrected cell line. We identified a sharp decrease in the expression of PPARGC1A, a master regulator of mitochondrial biogenesis. Nevertheless, it was not possible to determine the mechanism of suppression of PPARGC1A expression. Finally, our results suggest a possible link between the type of XPC mutation and PPARGC1A expression. This study unfolds new possible roles for XPC, aside from its established roles in genomic instability, in metabolic adaptation and redox imbalance towards tumour progression.

Les travaux expérimentaux présentés dans cette thèse associent deux pans de la physique de la matière condensée, avec d'un côté la physique des verres de spins et de l'autre la physique mésoscopique. Le verre de spins est un exemple emblématique de système désordonné et frustré, il se caractérise à basse température par un ordre magnétique non conventionnel, où le désordre magnétique apparaît gelé. De plus, celui-ci est considéré comme un système modèle pour étudier les verres en général et de ce fait, il a fait l'objet de nombreuses études expérimentales et théoriques. Après d'importants efforts de recherche, la description de l'état fondamental de ce système a abouti à deux approches très différentes. La première, donnée par la résolution non triviale du problème en champ moyen, met en avant un état fondamental composé d'une multitude d'états organisés et hiérarchisés. La deuxième approche, dite des "gouttelettes", se base quant à elle sur la dynamique hors équilibre d'un unique état. Cependant, en dépit de ces contributions, la compréhension de cette phase est loin d'être complète et la nature de l'état fondamental reste encore un débat ouvert. Dans un conducteur mésoscopique, le transport se fait de manière cohérente : les électrons gardent la mémoire de leur phase, ce qui permet d'observer des effets d'interférences électroniques. La motivation à la base de ce travail est d'utiliser ces effets d'interférences comme outil pour étudier le verre de spins. En effet, étant donné que les interférences électroniques dépendent intiment de la disposition du désordre statique du conducteur, le transport cohérent peut se révéler être une sonde microscopique très efficace pour étudier la configuration du désordre dans un conducteur. Bien que quelques expériences pionnières de transport cohérent existent dans des verres de spins, ce domaine de recherche n'a que très peu été exploré. Néanmoins, il a connu un récent renouveau grâce à des travaux théoriques qui montrent de quelle manière cette sonde est sensible au désordre magnétique gelé et comment elle peut fournir des informations sur la nature de l'état fondamental du verre de spins. Ainsi, ce travail de thèse expérimental présente l'implémentation de mesure de transport dans des verres de spins mésoscopiques. La première partie de l'étude est consacrée aux caractéristiques générales de transport classique et quantique de ces systèmes. Nous avons examiné les propriétés de la résistivité en fonction de la température et du champ magnétique et nous montrons que ces systèmes mésoscopiques possèdent bien des comportements attendus pour des verres de spins. Dans une deuxième partie, nous nous sommes intéressés au comportement de la magnétorésistance à bas. Nous avons mis en avant que celle-ci présente une forte hystérésis dont l'amplitude dépend fortement, de la température dans la phase vitreuse et de la vitesse de balayage du champ magnétique. Nous avons argumenté que ce comportement particulier traduit la mise hors équilibre du système et montrons comment la température et la vitesse de balayage du champ magnétique pilotent l'écart à l'équilibre. Dans cette partie, nous avons aussi examiné par des mesures de transport la relaxation du système vers l'équilibre, après l'avoir excité. Nous présentons également les propriétés de transport étonnantes que nous avons observées à bas champ, résultant de protocoles en températures et en champs magnétiques plus complexes. / The experiments presented in this thesis associate two fields of condensed matter physic, on the one hand with the spin glass physic and the other hand with the mesoscopic physic. The spin glass state is one of the most emblematic of disordered and frustred system and at low temperature, it is caracterized by an unconventionel order where the magnetic disorder is quenched. Moroever, it is considered as a model system for glasses in general and thereby it has been extensively studied, both experimentally and theoreticlly. After extensive research efforts, the description of fundamental state of the system has lead towards two well different approaches. The first, given by the mean field solution, highlights a fundamental composed of mulitple states organised and hierarchical. The second, called droplet model is based on the off--equilibrium dynamic of a unique ground state. However, despite these contributions, the understanding ot this phase is far from being complete and the nature of the ground state still remains an open question. In a mesoscopic conductor, the transport of electron is coherent: electrons keep the memory of their phase, so that one can observe interference effects. The main motivation of this work is to use these interference effects in order to to probe the spin glass state. Indeed, as electronic interference depends of the position of the static disorder, coherent transport can be a useful tool to study the configuration of the microscopic disorder. Althought few coherent transport experiments exist to probe the spin glass, this field of research has very little explored. Nevertheless, this area has been a revival thanks to theoritical work, showing how coherent transport is sensitived to the quenched disorder and how it may provide informations of the nature of fundamental state of spin glass. So, this experimental work deals with the implementation of transport measurements in mesoscopic spin glasses. The first part of the study is focused on the general charateristics of classical and quatum transport of these system. We have examined the resistivity as a function of the temperature and magnetic field and we show that these mesoscopic systems have a spin glass-like behaviour. In a second part, we have focused on the low field magnetoresistivity. We show that it presents a strong hysteresis, whose the amplitude is strongly depends, both of the temperature in the glassy phase and sweeping rate of the magnetic field. We argue that this particular behaviour is related to the out off-equilibrium of the system and we show how the temperature and the sweeping rate control the deviation to the equilibrium. In this part, we also examine by transport measurements how the system relaxes towards the equilibrium just after its excitation. In addition, we present surprinsing transport propreties that we observed, resulting of experimental protocols more sophisticated in temperatures and magnetic fields.

Espécies reativas de oxigênio (EROs) são normalmente e continuamente geradas em mitocôndrias, majoritariamente na cadeia de transporte de elétrons (CTE). Harman (1956, 1972 e 1992) teorizou que os radicais livres gerados nas mitocôndrias seriam a principal causa do envelhecimento. De fato, durante o envelhecimento é observado um desequilíbrio entre formação e remoção de EROs, que resulta em estresse redox. Essa condição favorece a formação de lesões oxidadas no DNA, acarretando em mutagênese ou morte celular. Diversos mecanismos moleculares cooperam para o reparo de DNA. Duas vias de reparo de DNA lidam com a maioria das lesões: o reparo por excisão de base (BER) e o reparo por excisão de nucleotídeos (NER). A via BER corrige pequenas modificações de bases que surgem de reações de desaminação, alquilação e oxidação. A via NER é mais versátil, reconhecendo lesões que distorcem a dupla hélice de DNA, como danos induzidos por luz UV e adutos volumos. Pacientes xeroderma pigmentoso (XP-A a XP-G) herdam mutações em um de sete genes que codificam proteínas envolvidas na via NER, ou em um gene que codifica uma polimerase translesão (XP-V). A doença é caracterizada por fotosensibilidade e incidência elevada de neoplasias cutâneas. A proteína XPC atua na etapa de reconhecimento da lesão de DNA na subvia de reparo global do genoma (GG-NER), e sua mutação dá origem aos sintomas clássicos de XP. Novas funções de XPC foram recentemente descritas: i) atuando como cofator na via BER auxiliando as DNA glicosilases OGG1, TDG e SMUG; ii) atuando como cofator transcricional de elementos responsivos a Oct4/Sox2, RXR e PPARα; e iii) na adaptação metabólica na transformação de queratinócitos. Então, propusemo-nos a investigar as relações entre XPC e a manutenção da integridade do DNA mitocondrial, a sensibilidade celular a estresse redox mitocondrial e possíveis alterações bioenergéticas e redox. Para tal, padronizamos um ensaio in vitro de cinética de incisão em DNA plasmidial a fim de investigarmos o possível papel de XPC no reparo de lesões oxidadas em mtDNA. Porém, nossos dados revelaram que XPC não se encontra em mitocôndrias. Apesar disso, células XP-C são mais sensíveis ao tratamento com azul de metileno (AM), antimicina A (AA) e rotenona (ROT), que geram estresse redox mitocondrial. A sensibilidade à AA foi completamente revertida em células corrigidas. Células XP-C apresentaram alterações quanto ao uso dos complexos mitocondriais, com diminuição da taxa de consumo de oxigênio (OCR) via complexo I e um aumento da OCR via complexo II, dependente da presença de XPC. Ademais, a linhagem XP-C apresentou um desequilíbrio redox mitocondrial com maior produção de EROs e menor atividade de GPx. O DNA mitocondrial de células XP-C apresentou níveis elevados de lesão e deleção, que no entanto não retornaram aos níveis encontrados em células selvagens na linhagem XP-C corrigida. Observamos uma acentuada diminuição da expressão de PPARGC1A, um importante regulador de biogênese mitocondrial. Contudo, não foi possível determinar o mecanismo de supressão da expressão de PPARGC1A. Por fim, identificamos que o tipo de mutação em XPC pode estar associado a expressão de PPARGC1A. Esse estudo abre novas possibilidade na investigação do papel de proteína XPC, à parte da instabilidade genômica, na adaptação metabólica e desequilíbrio redox em direção da progressão tumoral. / Mitochondria continuously produce reactive oxygen species (ROS), mainly at the electron transport chain. Harman (1956, 1972 e 1992) proposed that normal aging is driven by increased mitochondrially generated free radicals. Indeed, during the course of aging there is an increased imbalance between formation and removal of ROS, leading to redox stress. This condition favours the formation of oxidized DNA lesions, given rise to mutations and cell death. Several molecular mechanisms cooperates to repair the DNA. Two DNA repair pathways deal with the majority of lesions: base excision repair (BER) and nucleotide excision repair (NER). The BER pathway corrects small base modifications that arise from deamination, alkylation and oxidation reactions. The NER pathway is more versitile, recognizing helix-distorting lesions, such as UV-induced damage and bulky adducts. Xeroderma pigmentosum (XP-A to XP-G) patients inherit mutations in one of seven protein-coding genes involved in NER pathway, or in a gene coding a translesion DNA polymerase (XP-V). Photosensitivity and a thousand-fold increased in the risk of developing cutaneous neoplasms are the main clinical features of XP. XPC protein functions in the recognition step of global genome NER (GG-NER) sub-pathway, and mutations in this gene lead to classical XP symptoms. Recently, it has been described that XPC acts: i) as a cofactor in BER pathway through functional interaction with DNA glycosylases OGG1, TDG and SMUG1; ii) as coactivator in transcription at Oct4/Sox2, RXR and PPARα responsive elements; iii) in metabolic shift during keratinocytes transformation. Thus, we sought to investigate a possible role for XPC in the maintenance of mtDNA integrity, cellular sensitivity to mitochondrial redox stress and eventual bioenergetic and redox changes. For this purpose, we established an in vitro plasmid incision assay to investigate the possible role of XPC in the repair of oxidized lesions in mitochondrial DNA. However, our data revealed that XPC did not localized in mitochondria. Nonetheless, XP-C cells are more sensitive to methylene blue, antimycin A (AA) and rotenone treatment, which induce mitochondrial redox stress. The XP-C sensitivity to AA was completely reverted in XPC-corrected cells. XP-C cells presented altered usage of mitochondrial complexes, with decreased oxygen consumption rate (OCR) via complex I and increased OCR through complex II, an XPC-dependent phenomenon. Furthermore, the XP-C cell line showed mitochondrial redox imbalance with increased ROS production and decrease GPx activity. MtDNA from XP-C cells accumulate lesions and deletions, which, however, were found at similar levels in the corrected cell line. We identified a sharp decrease in the expression of PPARGC1A, a master regulator of mitochondrial biogenesis. Nevertheless, it was not possible to determine the mechanism of suppression of PPARGC1A expression. Finally, our results suggest a possible link between the type of XPC mutation and PPARGC1A expression. This study unfolds new possible roles for XPC, aside from its established roles in genomic instability, in metabolic adaptation and redox imbalance towards tumour progression.

Currently organic-inorganic hybrid perovskite solar cells (PSCs) is one kind of promising photovoltaic technology due to low production cost, easy fabrication method and high power conversion efficiency. Charge transport layers are found to be critical for device performance and stability. A traditional electron transport layer (ETL), such as TiO2 (Titanium dioxide), is not very efficient for charge extraction at the interface. Compared with TiO2, SnO2 (Tin (IV) Oxide) possesses several advantages such as higher mobility and better energy level alignment. In addition, PSCs with planar structure can be processed at lower temperature compared to PSCs with other structures. In this thesis, planar-structure perovskite solar cells with SnO2 as the electron transport layer are fabricated. The one-step spin-coating method is employed for the fabrication. Several issues are studied such as annealing the samples in ambient air or glovebox, different concentration of solution used for the samples, the impact of using filter for solutions on samples. Finally, a reproducible fabrication procedure for planer-structure perovskite solar cells with an average power conversion efficiency of 16.8%, and a maximum power conversion efficiency of 18.1% is provided.

Decline in cardiac pump function is a hallmark of aging where mitochondrial decay is an important underlying cause. Although certainly multifactorial in nature, both dysfunction of the machinery involved in the chemiosmotic process of energy transduction and lower capacity to maintain fatty acid-driven respiration are identified as intrinsic factors of mitochondrial decay in the aged myocardium. Age-associated destabilization of electron transport supercomplexes as a potential factor of mitochondrial decay in the rat heart. Defective operation of the electron transport chain (ETC) constitutes a key mechanism involved in the age-associated loss of mitochondrial energy metabolism. Nevertheless, the molecular events underlying inefficient electron flux that ultimately leads to higher superoxide appearance and impaired respiration are not fully known. As recent biophysical evidence shows that the ETC may form large macromolecular assemblies (i.e. supercomplexes) that disintegrate in certain pathologies (e.g. heart failure or Barth syndrome) reminiscent of aging, we investigated the hypothesis that alterations in supercomplexes are partly responsible for the age-related loss of cardiac ETC function. In this dissertation, age-associated changes in supercomplex organization and stability were investigated in subsarcolemmal (SSM) and interfibrillary (IFM) mitochondria isolated from cardiac tissue from young (3-5 months) and old (24-28 months) male Fischer 344 rats. Blue native-PAGE (BN-PAGE) analysis of digitonin-solubilized mitochondrial membranes coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to investigate supercomplex organization. Results show that both SSM and IFM display supercomplexes comprised of various stoichiometries of complexes I, III and IV (never complex II), which typically organize as high mass (1500-2300 kDa) assemblies containing up to four copies of complex IV (i.e. I���III���IV[subscript N]-type supercomplexes). Interestingly, analysis of IFM proteins showed that, in general, supercomplex levels declined by up to 15 % (p < 0.05) with age; however, different degrees of supercomplex deterioration were observed, depending on the particular supercomplex investigated. Supercomplexes of the highest molecular weights (i.e. 1900-2300 kDa), which were also composed of the most complex stoichiometries (i.e. I1III2IVN, N ��� 2), were primarily lost with age. In particular, I���III���IV���, I���III���IV��� and I���III���IV��� supercomplexes were found to decline by 13% (p < 0.05), 30% (p < 0.05) and 45% (p < 0.05), respectively, on an age basis. Therefore, the age-associated loss of supercomplexes in IFM stems from destabilization of the assemblies that comprise several copies of complex IV, which could partially limit proper electron transfer to O��� for its reduction, affecting mitochondrial respiratory capacity. In contrast to IFM, the aging defects of SSM supercomplexes appeared to be confined to the assembly comprised of only one copy of complex IV (I���III���IV���, 1700 kDa) (37% loss; p = 0.06), while the higher molecular weight supercomplex sub-types that were most affected in IFM (i.e. I���III���IV[subscript N], N ��� 2) were not significantly altered with age. Thus, the results from this dissertation indicate that mitochondria from different subcellular locations in the myocyte show different degrees of supercomplex destabilization in the aging rat heart. The more robust supercomplex deficits noted for IFM fit well with previous observations that electron transport characteristics of this subpopulation are more adversely affected with age than SSM. Although the underlying factor(s) of supercomplex deterioration are not fully known, the hypothesis that age-related alterations of certain constituents of the IMM (e.g. cardiolipin) may be important factors of supercomplex destabilization in cardiac mitochondria was investigated in this dissertation. To this end, LC-MS/MS characterization of supercomplex proteins and HPLC analysis of cardiolipin were used as approaches to elucidate potential factor(s) of supercomplex destabilization in the aging rat heart. Age-related alterations of cardiolipin levels and its acyl-chain content showed a strong parallel to the age-associated destabilization of supercomplexes. Specifically, cardiolipin levels declined by 10% (p < 0.05) in IFM, the mitochondrial subpopulation displaying the highest degree of supercomplex deterioration. In addition, the content of (18:2)���-cardiolipin, the predominant species in the heart, was found to decline by 50% (p < 0.05) on average in both populations of cardiac mitochondria. Therefore, the data presented in this dissertation indicate that changes in cardiolipin may be at least one of the factors involved in supercomplex destabilization in the aging heart. Age-related decline in carnitine palmitoyltransferase I (CPT1) activity as a mitochondrial lesion that limits fatty acid catabolism in the rat heart. Loss of fatty acid utilization, another intrinsic factor of mitochondrial decay in the aged myocardium, has been associated with age-related alterations in the activity of carnitine palmitoyltransferase 1 (CPT1), the rate-controlling enzyme for overall fatty acid ��-oxidation. Nevertheless, the exact molecular mechanism involved in the age-related loss of fatty acid-driven bioenergetics is not fully understood. In this dissertation, it was also investigated whether the aging lesion for fatty oxidation lies in a particular mitochondrial subpopulation or more generally results from cardiac decrements in L-carnitine levels. In order to clarify the role of each one of these factors, the effect of long-term dietary supplementation with the L-carnitine analogue, acetyl-L-carnitine (ALCAR), was also investigated. Results show that aging selectively decreases CPT1 activity in IFM by reducing enzyme catalytic efficiency for palmitoyl-CoA. IFM displayed a 28% (p < 0.05) loss of CPT1 activity, which correlated with a decline (41%, p < 0.05) in palmitoyl-CoA-driven state 3 respiration. Interestingly, SSM had preserved enzyme function and efficiently utilized palmitate. Analysis of IFM CPT1 kinetics showed both diminished V[subscript max] and K[subscript m] (60% and 49% respectively, p < 0.05) when palmitoyl-CoA was the substrate. However, no age-related changes in enzyme kinetics were evident with respect to L-carnitine. ALCAR supplementation restored CPT1 activity in heart IFM, but not apparently through remediation of L-carnitine levels. Rather, ALCAR influenced enzyme activity over time, potentially by modulating conditions in the aging heart that ultimately affect palmitoyl-CoA binding and CPT1 kinetics. In conclusion, this dissertation presents a characterization of age-associated alterations in the macromolecular organization of the IMM components that could partly explain the loss of mitochondrial oxidative capacity that affects the aging heart. In addition, the characterization of an age-related lesion of the controlling enzyme for ��-oxidation is presented as another important factor that limits mitochondrial function and energy metabolism in cardiac mitochondria. / Graduation date: 2013