L'écotoxicité de différentes nanoparticules de carbone (NPC) a été évaluée chez des organismes aquatiques, en particulier chez Xenopus laevis. Il a été montré que la surface des NPC est le paramètre le plus pertinent pour décrire l'inhibition de croissance chez le xénope, indépendamment de leur forme allotropique et de leur état de dispersion. L'induction des micronoyaux a aussi été étudiée chez le xénope, et l'oxyde de graphène (GO) s'est révélé génotoxique à faible dose, résultat corroboré par l'étude de l'expression des gènes. Les mécanismes de toxicité impliqués seraient notamment liés aux fonctions oxygénées de la particule. De plus, le GO a aussi entrainé de la génotoxicité chez Pleurodeles waltl. et de la tératogénicité, des retards de développement et de l'inhibition de croissance chez Chironomus riparius. La mise en interaction de ces organismes au sein d'un mésocosme a également conduit à l'observation de génotoxicité chez le pleurodèle en présence de GO. / The ecotoxicity of different carbon-based nanoparticles (CNPs) was assessed in freshwater organisms, especially in Xenopus laevis. The surface of the CNPs was shown to be the more relevant parameter to describe the growth inhibition in Xenopus, regardless of their allotropic form and their state of dispersion. Micronucleus induction was also studied in Xenopus and graphene oxide (GO) was found genotoxic at low dose. This result was in compliance with the study of genes expression. The involved toxicity mechanisms would be related to the oxidized functions of the CNP. Moreover, GO was also found responsible for genotoxicity in Pleurodeles waltl. and for teratogenicity, development delay and growth inhibition in Chironomus riparius. These organisms have finally been put together in a mesocosm, which has also led to genotoxicity in Pleurodeles in the presence of GO.

Philosophiae Doctor - PhD (Chemistry) / Supercapacitors, also known as ultracapacitors or electrochemical capacitors, are considered one of the most important subjects concerning electricity or energy storage which has proven to be problematic for South Africa. In this work, graphene oxide (GO) was supported with platinum, silver and copper nanoparticles anchored with dodecylbenzenesulphonic acid (DBSA) doped polyaniline (PANI) to form nanocomposites. Their properties were investigated with different characterization techniques. The high resolution transmission electron microscopy (HRTEM) revealed GO's nanosheets to be light, flat, transparent and appeared to be larger than 1.5 ?m in thickness. This was also confirmed by high resolution scanning electron microscopy (HRSEM) with smooth surfaces and wrinkled edges observed with the energy dispersive X-ray analysis (EDX) confirming the presence of the functional groups such as carbon and oxygen. The HRTEM analysis of decorated GO with platinum, silver and copper nanoparticles (NPs) revealed small and uniformly dispersed NPs on the surface of GO with mean particle sizes of 2.3 ± 0.2 nm, 2.6 ± 0.3 nm and 3.5 ± 0.5 nm respectively and the surface of GO showed increasing roughness as observed in HRSEM micrographs. The X-ray fluorescence microscopy (XRF) and EDX confirmed the presence of the nanoparticles on the surface of GO as platinum, silver and copper which appeared in abundance in each spectra. Anchoring the GO with DBSA doped PANI revealed that single GO sheets were embedded into the polymer latex, which caused the DBSA-PANI particles to become adsorbed on their surfaces. This process then appeared as dark regions in the HRTEM images. Morphological studies by HRSEM also supported that single GO sheets were embedded into the polymer latex as composite formation appeared aggregated and as bounded particles with smooth and toothed edges.

Bio-inspired or biomimetic chemistry deals with the replication of the nature’s fundamental processes, which can help in understanding the functioning of biological systems and develop novel applications. Although a large number of researchers worked towards the replication of natural synthetic pathways through biogenetic syntheses, enzyme mimicry by the small organic molecules and inorganic complexes emerged in leaps and bounds over the years. The development of biomimetic chemistry then continued in designing the molecules that can function like enzymes. And now, with the advent of nanotechnology, nanostructured materials have been shown to exhibit enzyme-like activities (nanozymes). Interestingly, the two distinct fields, biology and materials science, have been integrated to form an entirely new area of research that has captured a great attention. Along with the pronounced application of nanomaterials as drug delivery vehicles, anticancer agents, antimicrobials, etc., research is also focused on designing nanomaterials for the biomimetic applications. The thesis consists of five chapters. The first chapter provides a general overview of the recently discovered nanozymes that mimic heme-peroxidase, oxidase, superoxide dismutase, catalase, haloperoxidase and phosphatase. This chapter also deals with the nanozymes’ application in sensing and immunoassay, and as antioxidants, neuroprotective agents. The factors affecting the nanozymes’ activity and the challenges associated with them is also covered in this chapter. Chapter 2 is divided into two parts and it deals with the biomimetic properties of graphene-based materials. In part A, the remarkable peroxynitrite (PN) reductase and isomerase activities of hemin-functionalized reduced graphene oxide (rGO) is discussed. In part B, the activity of graphene oxide (GO) as peroxide substrate for the glutathione peroxidase (GPx) enzyme is discussed. In chapter 3, the oxidant material, V2O5, is shown to exhibit significant GPx-like antioxidant activity in its nano-form. Chapter 4 deals with the oxidase-like activity of MnFe2O4 nanooctahedrons for the antibody-free detection of major oxidative stress biomarker, carbonylated proteins. In chapter 5, the phosphotriesterase mimetic role of vacancy engineered nanoceria is discussed. instead of H2O2 for glutathione peroxidase (GPx) enzyme. As partial reduction of GO was observed when treated with GPx enzyme due to the fact that large sheet-like structures cannot be accessible to the active site, we studied the reaction with some GPx mimetics (Fig. 2). Varying the concentration of cofactor glutathione (GSH) required for the reaction, GPx mimic, ditelluride, could accomplish the reduction of GO following Michaelis-Menten kinetics. As the structure of GO is elusive and under active investigation, our study highlights the presence of peroxide linkages as integral part of GO other than hydroxyl, epoxy and carboxylic groups. This study also highlights an important fact that the modification of GO by biologically relevant compounds such as redox proteins must be taken into account when using GO for biomedical applications because such modifications can alter the fundamental properties of GO. Figure 2. The GO reductase and decarboxylase activities of GPx mimetic ditelluride compound, suggesting the presence of peroxide linkages on GO. In chapter 3, we have discussed about the novel antioxidant nanozyme that combats oxidative stress. During our attempts in the investigation of antioxidant nanozymes, we surprisingly noticed that the oxidant material, V2O5, shows significant GPx-like antioxidant activity in its nano-form. The Vn readily internalize in the cells and exhibit remarkable protective effects when challenged against reactive oxygen species (ROS). Although Vn has been shown to protect cells from ROS-induced damage, cells treated with bulk V2O5 and few vanadium complexes resulted in generation of ROS and severe toxicity. Detailed investigation on the mechanism of this interesting phenomenon Chapter 4 deals with the development of novel methodology for detection of biomarkers. Inspired by the use of antibodies and enzymes for detection of a specific antigen, we have shown for the first time that the nanozymes can entirely replace antibodies and enzymes in Enzyme-linked Immunosorbent Assays (ELISA). As a specific example, we focused on the antibody-free detection of chief oxidative stress biomarker, carbonylated proteins, as our target. To achieve this, we designed MnFe2O4 nanooctahedrons that can function as oxidase enzyme and form signaling point of detection. We functionalized MnFe2O4 nanooctahedrons with hydrazide terminating groups so that carbonylated proteins can be linked to nanozymes by hydrazone linkage (Fig. 4a). Treatment of various carbonylated proteins (hemoglobin (Hb), Myoglobin (Mb), Cytochrome c (Cyt c), RNase and BSA) coated in well plate with hydrazide-terminated MnFe2O4 nanooctahedrons and then with 3,3’,5,5’-tetramethylbenzidine substrate, resulted in instantaneous detection by well plate reader (Fig. 4b). Considering the challenges and difficulties associated with the conventional methods used to detect such modified proteins, this methodology opens up a new avenue for the simple, cost-effective, instantaneous and entirely antibody-free ELISA-type detection of carbonylated proteins. Our results provide a cumulative application of nanozymes’ technology in oxidative stress associated areas and pave a new way for direct early detection of post translational modification (PTM) related diseases. Figure 4. a) Nanozyme linked to the carbonylated protein coated on a plate through hydrazone linkage. b) General bar diagram showing detection of oxidized (carbonylated) proteins by nanozymes. Synopsis Figure 5. a) A cartoon view of surface of ceria showing vacancy. b) Zoomed portion of high resolution transmission electron microscopic image showing few vacancies on the surface of nanoceria. c) Catalytic mechanism of detoxification of paraoxon at the defect site. In the final chapter, chapter 5, we have discussed about the nanomaterial that can function as phosphotriesterase enzyme. Phosphotriesterase enzyme is a bacterial enzyme that is involved in the rapid hydrolysis of sarin gas-related deadly nerve agents such as paraoxon, parathion and malathion. When encountered with these orgnaophospatetriesters, living beings tend to undergo nerve shock to cause paralysis by inhibiting an extremely important enzyme called acetylcholine esterase. They are also known to cause severe oxidative stress problems and are associated with neurodegenerative disorders. Therefore, curbing the toxic effects and detoxification of these nerve agents is a world-wide concern and many research teams have focused their attention to address this important problem. Working on the development of nanozymes for important problems, we found that nanoceria, especially the vacancy engineered one (Fig. 5a,b), can serve as active mimic of phosphotriesterase enzyme in the presence of N-methylmorpholine (acting as a distal base histidine). Vacancy engineered nanoceria has been shown to catalyze the hydrolysis of high amounts of paraoxon quiet efficiently and within few minutes with very low activation energy and high kcat. Detailed mechanistic investigation revealed that the presence of both Ce(III) and Ce(IV) is very essential for detoxification activity (Fig. 5b). The vacancies on the surface of nanoceria, were the buried Ce(III) ions are directly exposed to the reaction environment, behave as hotspots or enzyme active sites for detoxification reaction (Fig. 5b).

My thesis is divided into five chapters, starting with a general introduction in first chapter and sample preparation and protein-NMR assignment techniques in second chapter. The remaining three chapters focus on three different areas/projects that I have worked on. Chapter 1: Introduction to nanomaterials and all the experimental techniques This chapter reviews different kinds of nanomaterials and their application utilized for protein-nanomaterial interaction in our study, along with the introduction to different spectroscopy and microscopy techniques used for the interaction studies. Starting with introduction of nanomaterials and all the experimental techniques, which constitute the arsenal for structural studies of the protein-nanomaterial interaction, different steps enroute to structural and dynamic interaction are outlined in detail. Chapter 2: Preparation and Characterization of Proteins used for nanomaterial interaction studies Proteins are generally of three kinds- globular (structured), intrinsically disordered and membrane bound. These proteins have different functions in living organisms and play a major role to maintain metabolism and other important factors. To probe protein-nanomaterial interactions, we have chosen different protein/peptides. This chapter describes the protocol/procedure used for purifying the proteins. For studying a globular protein, ubiquitin was chosen. Nanomaterial-IDP interaction was investigated using the intrinsically disordered central linker domain of human insulin like growth factor binding protein-2 (L-hIGFBP2). The hydrophobic membrane interacting part of the prion protein was chosen as a representative membrane protein. The characterization of the proteins by NMR spectroscopy is also described. Chapter 3: A nanomaterial based novel macromolecular crowding agent Carbon quantum dots (CQD) are nanomaterials with size less than 10 nm, first obtained in 2004 during purification of single-walled carbon-nanotubes. Since then CQDs have been used in a wide range of applications due to their low cost of preparation and favorable properties such as chemical inertness, biocompatibility, non-toxicity and solubility in aqueous medium. One of the applications of CQDs has been their use for imaging and tracking proteins inside cells, based on their intrinsic fluorescence. Further, quantum dots exhibit concentration dependent aggregation while retaining their solubility. Fluorescent carbon quantum dots (CQD) induce macromolecular crowding making them suitable for probing the structure, function and dynamics of both hydrophilic and hydrophobic peptides/ proteins under near in-cell conditions. We have prepared hydrophilic and hydrophobic quantum dots to see the crowding effect. After characterization of CQD, we tested the property of proteins with CQD and found that CQD behaves as a macromolecular crowding agent by mimicking near in-cell conditions. In our study, we have chosen a globular protein, an intrinsically disordered protein (IDP) and one hydrophobic membrane peptide. We have also compared the crowding property of CQD with ficoll which is widely used commercial crowding agent. The overall study tells that the CQD acts like crowding agent and can be used for the study of macromolecular crowding effect. This makes them suitable for structural and functional studies of proteins in near in-cell conditions. Chapter 4: Ubiquitin-Graphene oxide interactions Described here is the interaction of human ubiquitin with GO using NMR spectroscopy and other techniques such as Fluorescence spectroscopy, isothermal titration calorimetry (ITC), UV-Visible spectroscopy, dynamic light scattering (DLS), zeta potential measurements and transmission electron microscopy (TEM). The globular protein ubiquitin interacts with GO and undergoes a dynamic and reversible association-dissociation in a fast exchange regimen as revealed by NMR spectroscopy. The conformation of the protein is not affected and the primary interaction is seen to be electrostatic in nature due to the polar functional groups present on the protein and GO sheet surface. For the first time we have shown that the interaction between ubiquitin and GO is dynamic in nature with fast and reversible adsorption/desorption of protein from the surface of GO. This insight will help in understanding the mechanistic aspects of interaction of GO with cellular proteins and will help in designing appropriate functionalized graphene oxide for its biological application. Chapter 5: Section A: Interaction of an intrinsically disordered protein (L-HIGFBP2) with graphene oxide The interaction between intrinsically disordered linker domain of human insulin-like growth factor binding protein-2 (L-hIGFBP2) with GO was studied using NMR spectroscopy and other techniques such as isothermal titration calorimetry (ITC), dynamic light scattering (DLS), zeta-potential measurements. The study revealed that the disordered protein L-hIGFBP2 interacts with GO through electrostatic interaction and undergoes a dynamic and reversible association-dissociation in a fast exchange regime. The conformation of the protein is not affected. Section B: Stability of an Intrinsically disordered protein through weak interaction with Silver nanoparticles Using NMR spectroscopy and other techniques we probed the mechanism of L-hIGFBP2–AgNP interactions which render the IDP stable. The study reveals a mechanism which involves a relatively fast and reversible association–dissociation of L-hIGFBP2 (dynamic exchange) from the surface of AgNP. The AgNP–L-hIGFBP2 complex remains stable for more than a month. The techniques employed in addition to NMR include UV-Visible spectroscopy, dynamic light scattering (DLS), zeta potential measurements and transmission electron microscopy (TEM) to probe the protein-AgNP interaction here in this section.

[pt] A escassez de recursos de água doce está ameaçando nossa sociedade. A urbanização, a industrialização, o crescimento populacional e as alterações climáticas estão a representar um grande desafio para a segurança dos recursos hídricos humanos. Com base nessa situação crítica, os cientistas estão prestando cada vez mais atenção à dessalinização. Os métodos tradicionais de dessalinização empregam o processo de destilação. Esses métodos desempenham um papel importante no serviço de abastecimento de água em alguns locais carentes de água. No entanto, devido ao alto consumo de energia desses métodos, o preço da água produzida é elevado. Portanto, desenvolver novas tecnologias de dessalinização com baixo consumo de energia é de grande interesse e uma delas tem atraído a atenção dos pesquisadores, que é a osmose reversa (OR). O RO utiliza a membrana semipermeável como filtro, o que permite que a água ou moléculas relativamente pequenas passem por si mesmas, mas impede que as grandes moléculas ou íons penetrem. Esta tecnologia reduziu significativamente o consumo de energia em comparação com os métodos de destilação e rapidamente ocupa mais de 60 por cento da capacidade total de dessalinização instalada. O desempenho da tecnologia RO depende fortemente do material das membranas desempenha um papel importante. Nas últimas décadas, polímeros, por exemplo, poliamida e acetato de celulose, dominaram a indústria de RO de membrana semipermeável por sua boa eficiência de rejeição de sal e baixo custo de consumo de energia. No entanto, mesmo com as vantagens das membranas poliméricas, o custo final da água produzida ainda é alto. É por isso que os recursos de água doce ainda continuam sendo a preocupação. Desde a primeira vez que o grafeno foi produzido a partir do grafite, chamou a atenção de pesquisadores em todo o mundo por sua estrutura 2D ultrafina, excelente condutividade e transparência etc. Logo depois, o grafeno e seus derivados, como óxido de grafeno e óxido de grafeno reduzido, exibem potencial na dessalinização devido à sua estrutura 2D fina e expansibilidade. Este trabalho explora a possibilidade de aplicação de derivados de grafeno em um processo de dessalinização relativamente prático. No prsente projeto foram produzidos tanto GO (pelo método de Hammer) e RGO (a partir de aquecimento em atmosfera inerte) e de membranas a partir de acetato de celulose com GO e RGO. Ensaios de dessalinação também foram realizados para amostras produzidas variando de modo sistemático diferentes parâmetros de síntese de GO e RGO e de fabricação das membranas de acetato de celulose. / [en] Fresh-water resource scarcity is threatening our society. Urbanization, industrialization, population growth and climate change are making big challenge to human s water resource security. Based on this critical situation, scientists are paying more and more attention to desalination. Traditional desalination methods employ distillation process. These methods play an important role in water supply service in some water-lacked places. However, due to high energy consumption of these methods, the price of produced water is very high. Therefore, developing new desalination technologies with low energy consumption is of high interest and one of them has attracted researchers attention, which is reverse osmosis (RO).(1) RO utilizes the semi-permeable membrane as a filter, which allows the water or relatively small molecules pass through itself, but prevents the large molecules or ions from penetrate. This technology significantly reduced the energy consumption compared to the distillation methods and quickly takes more than 60 percent of the total installed desalination capacity.(2) The performance of RO technology strongly depends on the material of membranes plays an important role. In the past decades, polymers, for instance polyamide and cellulose acetate, dominate the semi-permeable membrane RO industry for their good salt rejection efficiency and low cost of energy consumption. However, even with the advantages of polymer membranes, the final cost of produced water is still high. That s why fresh-water resource still remain the concern. Since the first time that graphene was produced from graphite, it caught researcher’s attention all over the world for its ultra-thin 2D structure, excellent conductivity and transparency, etc. Soon after, graphene and its derivatives, such as graphene oxide and reduced graphene oxide, exhibit potential in desalination due to their thin 2D structure and expandability.(3) This work explores the possibility of application of graphene derivatives in a relatively practical desalination process. In the present project, GO (by the Hammer method), RGO (from heating in air atmosphere) and cellulose acetate membranes with GO and RGO were produced. Desalination tests were also performed for samples produced by systematically varying different parameters of GO, RGO and fabrication of cellulose acetate membranes.

A doença cardíaca valvar é um problema clínico que está presente em todos os países independente do nível de desenvolvimento econômico. Anualmente são realizadas mais de 275.000 cirurgias para substituição da válvula aórtica em todo o mundo e a estimativa é que esse número triplique até 2050. Dentre as próteses utilizadas destaca-se a bioprótese, que apesar de apresentar uma rejeição menor que a prótese mecânica, a sua durabilidade é reduzida em virtude de sua calcificação e posterior deterioração. A fim de aumentar a durabilidade da prótese confeccionada a partir do pericárdio bovino, neste estudo, foi incorporado o óxido de grafeno funcionalizado com amino-polietilenoglicol 4.000 Mn (OG-PEG-NH2 4.000 Mn) e 6.000 Mn (OG-PEG-NH2 6.000 Mn) ao biomaterial. A incorporação do óxido de grafeno funcionalizado ao pericárdio bovino foi realizada, após sua esterilização com radiação gama, de duas formas distintas: por adsorção física e por reação química. Os resultados demonstraram que houve a incorporação do OG-PEG-NH2 4.000 Mn, verificado pelo aumento dos parâmetros de ensaio de resistência analisados, como por exemplo, a resistência à deformação plástica permanente e também a carga máxima suportada. Em relação ao OG-PEG-NH2 6.000 Mn houve uma melhora na biocompatibilidade do nanomaterial, diminuindo a sua citotoxicidade. Sendo assim, pode-se concluir que o OG-PEG-NH2 4.000 Mn possui o potencial para melhorar a resistência mecânica e consequentemente aumentar a durabilidade do biomaterial. / Valvular heart disease is a clinical problem that is present in all countries regardless of the level of economic development. More than 275000 surgeries are performed annually for aortic valve replacement worldwide, and it is estimated that this number will triple by 2050. Among the prostheses used, the bioprosthesis stands out, which, although presenting a smaller rejection than the mechanical prosthesis, its durability is reduced by cause of its calcification and subsequent deterioration. In order to increase the durability of the prosthesis made from the bovine pericardium, the functionalized graphene oxide with amino-polyethylene glycol 4000 Mn (OG-PEG-NH2 4000 Mn) and 6000 Mn (OG-PEG-NH2 6000 Mn) to the biomaterial. The incorporation of functionalized graphene oxide to the bovine pericardium was performed, after its sterilization with gamma radiation, in two different ways: physical adsorption and chemical reaction. The results showed that the OG-PEG-NH2 4000 Mn was incorporated, verified by the increase of the resistance test parameters analysed, for example, the resistance to permanent plastic deformation and also the maximum supported load. In relation to OG-PEG-NH2 6000 Mn there was an improvement in the biocompatibility of the nanomaterial, reducing its cytotoxicity. Thus, it can be concluded that the OG-PEG-NH2 4000 Mn has the potential to improve mechanical strength and consequently increase the durability of the biomaterial.

Submitted by Rosivalda Pereira (mrs.pereira@ufma.br) on 2017-06-02T19:09:40Z No. of bitstreams: 1 RolffSpindola.pdf: 1912678 bytes, checksum: 00839e9b1b8a2b01a86770dd2561ec07 (MD5) / Made available in DSpace on 2017-06-02T19:09:40Z (GMT). No. of bitstreams: 1 RolffSpindola.pdf: 1912678 bytes, checksum: 00839e9b1b8a2b01a86770dd2561ec07 (MD5) Previous issue date: 2016-07-25 / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPQ) / Fundação de Amparo à Pesquisa e ao Desenvolvimento Científico e Tecnológico do Maranhão (FAPEMA) / A novel platform for electroanalysis of isoniazid based on multi-walled carbon nanotube oxide and graphene oxide composite as support to iron phthalocyanine (MWCNTO-GO/FePc) has been developed. The FePc/MWCNTO-GO modified electrode is sensible for isoniazid, decreasing substantially its oxidation potential to + 200 mV vs. Ag/AgCl. Electrochemical and electroanalytical properties of the FePc/MWCNTO-GO modified electrode were investigated by cyclic voltammetry, electrochemical impedance spectroscopy, scanning electrochemical microscopy, and amperometry. The sensor presents better performance in 0,1 mol L-1 phosphate buffer at pH 7,4. Under optimized conditions, a linear response range from 5 μmol L-1 to 476 µmol L-1 was obtained with a sensitivity of 0,0225 μA L μmol-1. The limits of detection and quantification for isoniazid determination were 0,56 μmol L-1 and 1,88 μmol L-1, respectively. The relative standard deviation for 10 determinations of 100 μmol L-1 isoniazid was 2,5%. The sensor was successfully applied for isoniazid determination in artificial body fluids. / No presente trabalho é apresentado o desenvolvimento de uma nova plataforma para eletroanálise de isoniazida baseada em (MWCNTO-GO/FePc). O eletrodo modificado com FePc/MWCNTO-GO diminuiu sensivelmente o potencial de oxidação da isoniazida para +200 mV vs. Ag/AgCl. As propriedades eletroquímicas e eletroanalíticas do eletrodo modificado com FePc/MWCNTO-GO foram investigadas por voltametria cíclica, espectroscopia de impedância eletroquímica, microscopia eletroquímica de varredura, e amperometria. O sensor apresentou melhor desempenho em tampão fosfato 0,1 mol L-1 com pH 7,4. Sob condições otimizadas uma faixa de resposta linear de 5 μmol L-1 a 476 µmol L-1 foi obtida com uma sensibilidade de 0,0225 μA L μmol-1. O limite de detecção e quantificação para a isoniazida foram determinados como sendo 0,56 μmol L-1 e 1,88 μmol L-1, respectivamente. O desvio padrão relativo para 10 determinações de 100 μmol L-1 de isoniazida foi de 2,5%. O sensor foi aplicado com sucesso para determinação de isoniazida em fluidos corporais artificiais.

A specific double-stranded DNA sensing system is of great interest for diagnostic and other biomedical applications. Zinc finger domains, which recognize double-stranded DNA, can be engineered to form custom DNA-binding proteins for recognition of specific DNA sequences. As a proof of concept, a sequence-enabled reassembly of TEM-1 β- lactamase system (SEER-LAC) was previously demonstrated to develop zinc finger protein (ZFP) arrays for the detection of a double-stranded bacterial DNA sequence. Here, we implemented the SEER-LAC system to demonstrate the direct detection of pathogenspecific DNA sequences present in E. coli O157:H7 on the lab-on-a chip. ZFPs customdesigned to detect shiga toxin in E. coli O157:H7 were immobilized on the cyclic olefin copolymer (COC) chip, which can function as a non-PCR based molecular diagnostic. Pathogen-specific double-stranded DNA was directly detected by engineered ZFPs immobilized on the COC chip, providing a detection limit of 10 fmole of target DNA in colorimetric assay. Therefore, in this study, we demonstrated a great potential of ZFP arrays on the COC chip for further development of a simple and novel lab-on-a chip technology for detection of pathogens. Antibiotic resistance is a serious, and rapidly growing global threat. Here, we designed a novel screening method to detect antibiotic resistance genes (ARGs) in bacteria using a graphene oxide-based biosensor utilizing engineered ZFPs. Two-dimensional graphene oxide (GO) sheet possesses unique electronic, thermal, and mechanical properties. The quenching ability of GO can create novel methods for detection of biomolecules. Our approach utilizes quenching of fluorescence signal by GO in the absence of target ARGs, but restoring the signal in the presence of target ARGs. Quantum dot (QD)- labeled ZFP can bind to GO via stacking interactions of aromatic and hydrophobic residues in conjunction with hydrogen bonding interaction between hydroxyl or carboxyl groups of GO and hydroxyl or amine groups of the protein. Due to fluorescence resonance energy transfer (FRET) between QD and GO when they are in close proximity, fluorescence signal of QD-labeled ZFP is expected to be quenched. In the presence of target DNA, the bound DNA-protein complex is released from GO, restoring the fluorescence signal.

L’oxyde de graphène est un nanomatériau prometteur grâce à ses caractéristiques physicochimiques. Cependant, jusqu’à aujourd’hui, sa composition exacte reste encore inconnue. Ceci est dû à la complexité et au caractère non-stoechiométrique de ce matériau. Nous avons commencé par étudier sa composition de surface et sa réactivité. Nous avons utilisé des échantillons synthétisés de manière différente pour explorer la relation entre la méthode de synthèse et la composition de surface. En outre, nous avons préparé un dérivé fonctionnalisé avec un agent chélatant de radionucléides pour étudier sa biodistribution et l’impact de la taille latérale.Par la suite, nous avons essayé plusieurs stratégies de multi-fonctionnalisation. L’avantage est de pouvoir combiner différentes propriétés. Nous avons observé que, souvent après la fonctionnalisation, la dispersabilité de l’oxyde de graphène diminue. Ainsi, nous avons développé un échantillon fonctionnalisé par un polymère soluble dans l’eau. Enfin, nous avons exploré et amélioré les méthodes de caractérisation de l’oxyde de graphène. Une caractérisation approfondie par différentes techniques est fondamentale pour comprendre les modifications que le matériau a subies. / Graphene oxide is a promising nanomaterial thanks to its physicochemical characteristics. However, until today its exact composition remains still unknown. This is due to the complexity and non-stoichiometric character of this material.We started by investigating the surface composition of graphene oxide and its reactivity. We used differently synthesized samples to explore the relationship between the synthesis method and the surface composition. Furthermore, we functionalized graphene oxide with a chelating agent of radionuclides to study its biodistribution, and the impact of the lateral size. Afterwards, we tried different strategies for multifunctionalization with the aim to combine different properties. We observed that the dispersibility of graphene oxide often decreased after functionalization. Thus, we developed a highly water-stable graphene oxide sample by grafting awater-soluble polymer on its surface. Finally, we explored and improved the characterization methods for graphene oxide. Athorough investigation using different characterization techniques is fundamental to understand the modifications that the material underwent.

>Magister Scientiae - MSc / These days there are deep concerns over the environmental consequences of the rate of consumption of energy from non-renewable sources because of the accelerated increase in greenhouse effect. There is, therefore, increasing interest in research activities on renewable energy systems (e.g., supercapacitors, batteries, fuel cells and photovoltaic cells) and their materials. Supercapacitor materials have attracted much attention because of their high energy storage capacity, large surface area, high specific power density (watts/kg) and low cost. The development of advanced supercapacitor devices requires active electrode materials with high storage capacity and dispensability. Graphene oxide-dendritic star copolymer nanocomposites are fascinating as electrode materials, both scientifically and technologically, due to their exceptional properties, including light weight and high potential. / 2020-08-31