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Raman spectroscopy, together with multivariate statistical analyses, has proven to be a near real-time analytical technique capable of phenotyping cells, tissues and organs. This dissertation will show exclusively the application of the Raman spectroscopy phenotypic profiling method to; (i) microbial toxicity, (ii) ex-vivo organ perfusion, and (iii) subcellular location targeting. Real-time analytical methods for monitoring living biological systems will enable study of the physiological changes associated with growth, genetic manipulations, and adverse environmental conditions. Most existing analytical methods (NMR exempt), though highly accurate, must be performed off-line and most require destruction of the studied sample. These attributes make these methodologies less desirable to the study of physiological changes of cells, tissues, and organs. In this work, Raman spectroscopy has been identified and shown to be a good candidate for real-time analysis mainly because it can be performed: (i) in near real-time, (ii) non-destructively and with minimal sample preparation, (iii) through a glass barrier (i.e., can be performed in situ), and (iv) with minimal spectral interference from water. Here, Raman spectroscopy was used in combination with multivariate statistics to analyze the differing toxic effects of 4-C chain alcohols on E. coli. Good correlations were established between Raman spectra and off-line analytical techniques used to measure: (i) saturated, unsaturated, and cyclopropane fatty acids; (ii) amino acid composition of total protein; and (iii) cell membrane fluidity. Also, Raman 'fingerprint' analysis was used to discriminate among different phenotypic responses of cells. In addition, this methodology was applied to analyze perfusates of organs maintained by the VasoWave® organ perfusion system. Raman fingerprints can be used to assess organ health, and it is believed this data can be used to inform decisions such as whether or not to transplant an organ. Finally, molecular biology techniques were used to design and produce specific protein targets harboring a silver binding domain fusion, which upon release migrate to specific subcellular locations. By employing the related technique of surface-enhanced Raman scattering (SERS), which produces a highly amplified Raman signal in the presence of metallic nanoparticle substrates (e.g., silver nanoparticles), different regions of the E. coli cell structure were studied. The target regions studied by the technique included: (i) outer cell membrane, (ii) periplasm, and the (iii) cytoplasm. / Ph. D.

<p>With 2.1 million new cases, breast cancer is the most common cancer in women. Triple negative breast cancer (TNBC), which is 15-20% of these breast cancer cases is clinically negative for expression of estrogen and progesterone receptors (ER/PR) and human epidermal growth factor receptor 2 (HER2) receptors<a>.</a> It is characterized by its unique molecular profile, aggressive behavior, distinct patterns of metastasis, and lack of targeted therapies. TNBCs utilize glycolysis for growth, proliferation, invasiveness, chemotherapeutic resistance and hence has poor therapeutic response. There is an urgent need for novel/alternate therapeutic strategies beyond current standard of treatment for this subset of high-risk patients. Electrical pulse-based chemotherapy, known as electrochemotherapy (ECT) could be a viable option for TNBC therapy. ECT involves the local application of precisely controlled electrical pulses to reversibly permeabilize the cell membrane for enhanced uptake. ECT can increase the cytotoxicity of the chemotherapeutics up-to 1000 times, facilitating a potent local cytotoxic effect. </p> <p>The high cost and severe side-effects of conventional chemotherapeutics motivate the application of effective natural compounds. Combining electrical pulses with natural compounds will enhance the treatment efficacy. This dissertation focuses on curcumin, the yellow pigment of natural herb turmeric, that has been used for over 5000 years for its excellent anticancer properties. Previous studies have demonstrated the effectiveness of curcumin for treating multiple cancers, including TNBC, with limited side effects. The potency of curcumin can be enhanced further by combining it with ECT to provide an attractive and cost-effective alternative for TNBC treatment. </p> <p>Towards this we studied the effect of ECT with curcumin on MDA-MB-231 cell line, a human adenocarcinoma epithelial TNBC cell line. We performed various assays, including cell viability, colony forming, cell cycle, apoptosis, H₂O₂ reactive oxygen species (ROS), immunoblotting, real time quantitative PCR (qPCR), and cellular metabolites detection to study the impact of ECT with curcumin on MDA-MB-231 cells. In addition, to better understand the underlying mechanisms, we used high throughput, label-free quantitative proteomics. While several studies have attempted to define the mechanism of action of curcumin on cancer cells, little is known on the action mechanism of the curcumin delivered with electrical pulses. This work unravels the molecular mechanism behind the enhanced effects observed under the ECT-based curcumin therapy in TNBC cells, employing a high-throughput, quantitative, label-free mass spectroscopy-based proteomics approach. The proteomics approach provides information on the thousands of cellular proteins involved in the cellular process, allowing a comprehensive understanding of the electro-curcumin-therapy mechanism. Similar studies were also performed for ECT with cisplatin to compare the efficacy of the electro-curcumin-therapy to the standard stand-alone cisplatin-based therapy.</p> <p>Our results revealed a switch in the metabolism from glycolysis to mitochondrial metabolic pathways. This metabolic switch caused an excessive production of H₂O₂ ROS to inflict apoptotic cell death in MDA-MB-231 cells, demonstrating the potency of this ECT based curcumin therapy. These results encourage further studies to extend the application of ECT for clinical practice.</p>

Cette thèse s’intéresse à la synthèse biosourcée de métallophosphates poreux pour trouver des solutions à l’épuisement des réserves naturelles de sources de phosphore qui servent habituellement à l’élaboration des réactifs inorganiques phosphorés (ex. H3PO4). Ceux-ci sont notamment utilisés pour la synthèse des métallophosphates poreux. Ces derniers possèdent des propriétés importantes dans les domaines de la catalyse, de l’adsorption et de la séparation. Toutefois il existe d’autres domaines peu étudiés tels que le biomédical et l’environnement où ces matériaux sont aussi utilisés. Au cours de cette thèse, la synthèse de zincophosphates et d’aluminophosphates poreux biosourcés a été réalisée à partir de différentes biomolécules (la caséine et les nucléotides) par la voie hydrothermale. Plusieurs paramètres de synthèse ont été étudiés (pH, température de synthèse, concentration des réactifs, influence des conditions de calcination ...). Les solides ont été caractérisés à l’aide de différentes techniques instrumentales (DRX, MEB, RMN31P, manométrie de N2, spectrofluorimétrie ...). Les propriétés antibactériennes et cytotoxiques des zincophosphates éventuellement enrichis en nanoparticules d’argent synthétisées à partir de caséine ont été évaluées sur la souche bactérienne Escherichia coli K 12 et des cellules ostéoprogénitrices STRO1A+ respectivement. Ces travaux situés à l’interface de la chimie des matériaux et de la biologie permettent d’envisager l’application des nouveaux métallophosphates synthétisés dans les domaines émergents du biomédical et de l’environnement. / This thesis work focuses on the biosourced synthesis of metal phosphates in order to find solutions to the depletion of the natural phosphorus resources that are usually used for the preparation of the phosphorus-containing inorganic reagents (i.e., H3PO4). Hydrothermal synthesis of metal phosphates involved them for example. Porous metal phosphates have important properties in catalysis, adsorption and separation fields. However, there are also less explored domains such as biomedical and environmental ones where they are further used. During this thesis, the synthesis of biosourced porous zinc phosphates and aluminum phosphates has been carried out using different biomolecules (casein and nucleotides) by hydrothermal method. Many synthesis parameters were studied (pH, synthesis temperature, reagents concentration, influence of calcination conditions ...). Solids were characterized using different techniques (XRD, SEM, 31P NMR, N2 adsorption, spectrofluorimetry ...). The antibacterial and cytotoxic properties of zinc phosphates optionally enriched in silver nanoparticles synthetized with casein were evaluated on the bacterial strain Escherichia coli K 12 and STRO1A+ osteoprogenitor cells respectively. This work at the interface of materials chemistry and biology makes possible to consider the application of the prepared new metal phosphates for emerging biomedical and environment fields.

Made available in DSpace on 2017-07-24T19:38:14Z (GMT). No. of bitstreams: 1 RAFAELA D OLIVEIRA.pdf: 2756499 bytes, checksum: e1e60d222b6dbe62066658f22ea4de0b (MD5) Previous issue date: 2014-08-22 / In this work, starch-stabilized silver nanoparticles (AgNPs-Am) were synthesized by reduction of AgNO3 using the NaBH4. The temperature and concentration of reagents of the synthesis were optimized. The formation of AgNPs-Am was monitored by UV-Vis spectroscopy and dynamic light scattering (DLS). The optimum conditions found for the AgNPs-Am synthesis were starch 0.6 % (w/v), 3.6 x 10-3 mol L-1 NaBH4, 0.9 x 10-3 mol L-1 AgNO3 and synthesis in bath ice. The average size of the AgNPs-Am was between 21 and 77 nm. Transmission electron microscopy (TEM) confirmed the AgNPs-Am formation inside and outside of the starch chains, however the smaller sizes were referred to the NPs stabilized by starch. Measurements of Potential zeta indicated stability of the particles, confirmed by DLS monitoring that demonstrated low agglomeration of NPs in a period of 115 days. For the characterization of AgNPs-Am it was also used infrared spectroscopic (FTIR) and X-ray diffraction (XRD). The AgNPs-Am were applied as polyanion for the construction of films by Layer-by-Layer technique (LbL), alternating with polycation 3-n-propylpyridinium-silsesquioxane (SiPy+Cl-). In order to obtain the films, pH and immersion time of the polyelectrolytes were optimized, as well as the concentration of SiPy+Cl-, monitoring the deposition by UV-Vis. The optimum parameters were immersion time 240 seconds, 2 mg/mL SiPy+Cl- pH 6.5 and AgNPs-Am pH 9.0. Atomic force microscopy (AFM) images showed that film thickness increases linearly and the roughness decrease with the bilayers number. FTIR spectra and Raman confirmed the interaction between the polyelectrolytes in the assembly of LbL films. The LbL films with architecture (SiPy+Cl-/AgNPs-Am)n (n = bilayers number) were applied as modified electrodes for iodine detection, using differential pulse voltammetry (DPV). It was verified that interaction of the components in the LbL films improved the current intensity. The film (SiPy+Cl-/AgNPs-Am)n (n=5) showed better current response in phosphate buffered saline (PBS) 0.1 mol L-1 pH 7.0. The instrumental parameters Epulse, and tpulse were optimized by 23 factorial design. It was verified a significant effect for third order interaction for the intensity of iodine redox peak currents, so the instrumentals parameters were evaluated together. The results of optimization were tpulse = 0.05 s, = 40 mV/s e Epulse = 50 mV. Accordingly, the modified electrode obtained a linear response for iodine concentrations ranging from 4.34 x 10-5 to 3.47 x 10-4 mol L-1 (R=0,9936) and from 4.40 x 10-4 to 4.24 x 10-3 mol L-1 (R=0,9938). It was obtained limit of detection (LOD) 5.56 x 10-6 and 1.51 x 10-5 mol L-1 and for limit of quantification (LOQ) 1.85 x 10-5 and 5.04 x 10-5 mol L-1, respectively. The AgNPs-Am synthesized in this work also acted as colorimetric sensor for iodine, with three regions of linearity. Two analytical curves were obtained for iodine concentration range from 2.40 x 10-7 to 9.50 x 10-7 mol L-1 and from 2.40 x 10-6 mol L-1 to 1.60 x 10-5 mol L-1. It was obtained a LOD of 1.71 x 10-8 and 1.06 x 10-6 mol L-1 and LOQ of 5.69 x 10-8 to 3.55 x 10-6 mol L-1, respectively for each range. Color variations obtained in these concentrations correspond to interaction between iodine and silver, which were monitored by UV-Vis band showed in 410 nm. At concentrations above 5.50 x 10-5 mol L-1 there is the appearance of blue color, absorbance in 600 nm, corresponding to the interaction between starch and iodine. It was also obtained a linear relationship for iodine concentration from 5.50 x 10-5 mol L-1 to 9.50 x 10-5 mol L-1. For this concentration range, LOD and LOQ were respectively 1.37 x 10-6 and 4.58 x 10-6 mol L-1. The results presented confirm the potential use of AgNPs-Am for iodine detection, both for the modification of electrodes for electrochemical determination as a colorimetric sensor. / Neste trabalho foram sintetizadas nanopartículas de prata estabilizadas em amido (AgNPs-Am). A síntese foi realizada por redução do sal AgNO3 utilizando o NaBH4. A temperatura de síntese e concentração dos reagentes foram otimizadas, a formação das AgNPs-Am foi monitorada por espectroscopia na região do UV-Vis e medidas de espalhamento dinâmico da luz (DLS). As condições ótimas encontradas para síntese de AgNPs-Am foram amido 0,6 % (m/v), 3,6.10-3 mol.L-1 de NaBH4, 0,9.10-3 mol.L-1 de AgNO3 e síntese em banho de gelo. O tamanho médio das AgNPs-Am foi entre 21 e 77 nm. Imagens de microscopia eletrônica de transmissão (TEM) confirmaram que houve formação das AgNPs no interior e fora das cadeias de amido, sendo os menores tamanhos referentes às NPs estabilizadas pelo amido. Medidas de Potencial zeta indicaram estabilidade das NPs, confirmada por acompanhamento DLS que demonstrou baixa aglomeração em um período de 115 dias. Para caracterização das AgNPs-Am utilizou-se também espectroscopia na região do infravermelho (FTIR) e difração de raios X (XRD). As AgNPs-Am foram utilizadas como poliânion para construção de filmes pela técnica Layer-by-Layer (LbL), alternando com policátion 3-n-propilpiridínio-silsesquioxano (SiPy+Cl-). Para construção dos filmes, pH e tempo de imersão dos polieletrólitos foram otimizados, assim como concentração de SiPy+Cl-, monitorando a deposição por UV-Vis. Os parâmetros ótimos foram tempo de imersão 240 segundos, solução de 2 mg/mL de SiPy+Cl- pH 6,5 e AgNPs-Am pH 9,0. Imagens de microscopia de força atômica (AFM) mostram que a espessura dos filmes aumenta linearmente e a rugosidade decresce com o número de bicamadas. Espectros FTIR e Raman confirmaram a interação entre os polieletrólitos na montagem dos filmes. Os filmes LbL com configuração (SiPy+Cl-/AgNPs-Am)n (n = número de bicamadas) foram aplicados como eletrodos modificados para detecção de iodo, utilizando-se voltametria de pulso diferencial (VPD). Verificou-se que a interação dos componentes no filme LbL melhorou a intensidade de corrente. O filme (SiPy+Cl-/AgNPs-Am)n (n=5) apresentou melhor resposta de corrente em eletrólito suporte tampão fosfato salino (PBS) 0,1 mol.L-1 pH 7,0. Os parâmetros instrumentais Epulso, e tpulso foram otimizados por planejamento fatorial 23. Verificou-se efeito significativo para interação de terceira ordem para intensidade de corrente do pico de redução do iodo, portanto os parâmetros instrumentais foram avaliados em conjunto. Os resultados da otimização foram tpulso = 0,05 s, = 40 mV/s e Epulso = 50 mV. Nestas condições, o eletrodo modificado obteve resposta linear para concentrações de iodo que variam de 4,34.10-5 a 3,47.10-4 mol.L-1 (R=0,9936) e de 4,40.10-4 a 4,24.10-3 mol.L-1 (R=0,9938). Respectivamente, obteve-se limite de detecção 5,56.10-6 e 1,51.10-5 mol.L-1 e para o limite de quantificação 1,85.10-5 e 5,04.10-5 mol.L-1. As AgNPs-Am sintetizadas neste trabalho também atuaram como sensor colorimétrico de iodo, com três regiões de linearidade. Duas curvas analíticas foram construídas para concentrações de iodo de 2,40.10-7 a 9,50.10-7 mol.L-1 e de 2,40.10-6 mol.L-1 a 1,60.10-5 mol.L-1. Obteve-se LD de 1,71.10-8 e 1,06.10-6 mol.L-1 e LQ de 5,69.10-8 e 3,55.10-6 mol.L-1, respectivamente para cada intervalo. As variações de cores obtidas nestas concentrações correspondem a interação entre o iodo e a prata, as quais foram acompanhadas pela banda UV-Vis apresentada em 410 nm. Em concentrações acima de 5,50.10-5 mol.L-1 observa-se o aparecimento de coloração azul, absorbância em 600 nm, correspondente a interação entre amido e iodo. Obteve-se linearidade para concentrações de 5,50.10-5 mol.L-1 a 9,50.10-5 mol.L-1. Para este intervalo de concentração, os LD e LQ são respectivamente 1,37.10-6 e 4,58.10-6 mol.L-1. Os resultados apresentados confirmam a potencialidade do uso das AgNPs-Am para detecção de iodo, tanto na modificação de eletrodos para determinação eletroquímica, quanto o uso como sensor colorimétrico.

CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / As nanopartÃculas metÃlicas apresentam inÃmeras aplicaÃÃes, como na construÃÃo de sensores, microeletrÃnica, catÃlise, aÃÃo bactericida, cÃlulas fotovoltaicas, entre outras. Dentre elas, a nanopartÃculas de prata (NPAg) tem ganhado destaque, em especial pela a sua atividade bactericida e por exibirem propriedades Ãpticas e eletromagnÃticas diferentes das observadas no metal agregado. VÃrias metodologias de sÃntese de nanopartÃculas de prata jà foram desenvolvidas. Para substituir os mÃtodos clÃssicos, que se baseiam na reduÃÃo dos Ãons prata por boroidreto de sÃdio ou citrato de sÃdio. A sÃntese verde, uma dessas novas metodologias, propÃe o uso de extrato de plantas, aÃÃcares, microrganismos e polissacarÃdeos para a reduÃÃo dos Ãons prata e estabilizaÃÃo das nanopartÃculas obtidas. O objetivo deste trabalho à propor a sÃntese verde de nanopartÃculas de prata, usando o extrato aquoso de sementes de girassol como agente redutor e soluÃÃo de galactomanana da fava danta como agente estabilizante. As NPAg foram sintetizadas adicionando-se 500 &#956;L de soluÃÃo de AgNO3 a 20 mL de soluÃÃo de galactomanana 0,032% (m/v) e 5 mL do extrato aquoso. As sÃnteses foram conduzidas a temperatura ambiente, 50, 70 e 90 ÂC, utilizando soluÃÃes 10 e 100 mmol/L de AgNO3 e soluÃÃo de galactomanana 0,032 %. O extrato aquoso das sementes de girassol liofilizado, apresentou um teor de umidade de 11,7%, um teor de proteÃnas de 15,5% e um resÃduo a 900ÂC de 12,8%. A formaÃÃo das nanopartÃculas foi confirmada por espectroscopia de absorÃÃo na regiÃo do UV-Vis, pela banda de ressonÃncia de plasmon na regiÃo de 400 nm. Os sistemas reacionais, com maior concentraÃÃo de Ag+, mostraram-se mais eficiente que os sistemas com menor concentraÃÃo desse Ãon. Entretanto, as NPAg formadas apresentaram maiores valores de &#955;mÃx (lambda mÃxima) e LMA (largura de banda à meia altura), indicando maiores tamanhos e maior polidispersividade das mesmas. O aumento da temperatura favorece a formaÃÃo das NPAg para todos os sistemas, inclusive os sistemas com menor concentraÃÃo de Ãons prata. Os coloides obtidos em presenÃa de soluÃÃo de galactomanana apresentaram bandas de plasmon simÃtricas, bem definidas e estreitas, indicando a galactomanana como estabilizante na formaÃÃo das NPAg. Avaliou-se a estabilidade temporal dos coloides por anÃlise de UV-Vis, os resultados demonstraram que os coloides obtidos sÃo estÃveis. O poder redutor do extrato das sementes de girassol nÃo sofreu alteraÃÃo com o aquecimento prÃvio a temperatura de 90 ÂC e nem apÃs processo de liofilizaÃÃo e resuspensÃo, porÃm o poder redutor do extrato diminuiu apÃs uma partiÃÃo com acetato de etila e apÃs um processo de diÃlise. O MET mostrou que as nanopartÃculas de prata formada apresenta formato esfÃrico com pouca polidispersÃo e diÃmetro na faixa de Â17 nm. / Metallic nanoparticles have several applications, such as in biosensor, microelectronics, catalysis, bactericidal action and photovoltaic cells. Silver nanoparticles (NPAg) has gained prominence, especially for its bactericidal activity and exhibit different optical and electromagnetic properties. Several methods for the synthesis of silver nanoparticles have been developed to replace traditional methods, which are based on the reduction of silver ions by sodium borohydride or sodium citrate. The green synthesis proposes the use of plant extracts, sugars, polysaccharides, and microorganisms for the reduction of silver ions and stabilization of the nanoparticles obtained. The objective of this work is to propose the green synthesis of silver nanoparticles using aqueous extract of sunflower seeds as a reducing agent and galactomannan from fava danta, as a stabilizer agent. NPAg were synthesized by adding 500 &#956;l of a solution of AgNO3 and 20 mL of galactomannan 0.032 % (w/v) and 5 mL of aqueous extract. The syntheses were carried out at room temperature, 50, 70 and 90  C, using 10 to 100 mmol/L AgNO3 solution and 0.032% solutions of galactomannan. The aqueous extracts of sunflower seeds, when dried showed a moisture content of 11.7 %, protein content of 15.5% and a residue at 900 C of 12.8 %. All colloid synthesized were analyzed by absorption spectroscopy in the UV-Vis confirming the formation of the band NPAG the appearance of surface plasmon resonance (SPR) in the region of 400 nm. The reaction systems with higher concentrations of Ag+, were more efficient than systems with lower concentration of this ion. However the NPAg formed, showed higher &#955;mÃx (maximum lambda) and LMA (bandwidth at half maximum), indicating larger sizes and higher polydispersity of the same. Raising the temperature favors the formation of NPAG to all systems, including systems with lower concentrations of silver ions. Colloids obtained in the presence of galactomannan solution showed bands of symmetric plasmon, well defined and narrow, indicating the galactomannan as a stabilizer in the formation of NPAg. The temporal stability of the colloids was evaluated by UV-Vis analysis, and the results obtained showed that the colloids are stable. The reducing power of the extract of sunflower seeds did not change with the preheating (90  C) and even after lyophilization and resuspension process, but the reducing power of the extract decreased after a partition with ethyl acetate and after a dialysis process. TEM analysis showed that silver nanoparticles presents spherical shape with low polydispersity and diameter in the range of Â17 nm.

Nous avons étudié la chimie de surface de carbones poreux fonctionnalisés avec des oximes et amidoximes d’une part, et avec de la guanidine d’autre part. Nous avons axé ces travaux sur la compréhension et la quantification de ces fonctionnalisations par analyse thermique et plus particulièrement par thermo-désorption programmée en température couplée à la spectroscopie de masse (TPD-MS). Par la suite, nous avons évalué l’intérêt de la fonctionnalisation par la guanidine des carbones mésoporeux pour deux applications : le piégeage du CO2 et la capacité à synthétiser et stabiliser des nanoparticules métallique. Pour la première application, des tests d’adsorption du CO2 à 0°C et 20°C à 1 bar ont été réalisés sur un carbone mésoporeux oxydé sous air en présence ou non de guanidine. L’intensité des interactions (physisorption versus chimisorption) a été sondée par des calculs de chaleurs isostériques d’adsorption. Pour la seconde applications, nous avons utilisé des carbones mésoporeux oxydés en présence ou non de guanidine pour synthétiser des nanoparticules d’argent de taille inférieure à 2 nm et relativement monodisperses en taille. Des particules bimétalliques d’AgCu ont également été obtenues au sein de ces matrices carbonées mésoporeuses imprégnées par la guanidine. On a pu mettre ici en évidence un rôle bénéfique de la guanidine sur le taux de cuivre réduit. Des suivis thermiques par MET in situ ont également été réalisés afin d’évaluer la stabilité thermique de ces nanoparticules et de mieux appréhender l’intérêt de la fonctionnalisation sur les phénomènes de frittage de ces nanoparticules. Finalement, des tests catalytiques pour l’époxydation sélective du styrène ont aussi été effectués sur le matériau Ag/C. Les résultats préliminaires sont très prometteurs pour un procédé de préparation de catalyseur très simple à mettre en œuvre. / In this work, we studied the surface chemistry of porous carbons functionalized firstly with oximes and amidoximes and then with guanidine. We focused this work on the understanding and quantification of these functionalization with thermal analysis and more particularly by Temperature Programmed Desorption/Mass Spectrometry (TPD-MS). Subsequently, we evaluated the interest of guanidine functionalization of mesoporous carbons for two applications : the capture of CO2 and the ability to synthesize and stabilize metal nanoparticles. For the first application, CO2 adsorption tests at 0 ° C. and 20 ° C. at 1 bar were carried out on a mesoporous carbon oxidized under air in the presence or absence of guanidine. The intensity of interactions (physisorption versus chemisorption) was probed by isosteric adsorption heat calculations. For the second applications, we used oxidized mesoporous carbons with or without guanidine to synthesize silver nanoparticles less than 2 nm in size and relatively monodisperse in size. Bimetallic AgCu particles were also obtained from these mesoporous carbon matrices impregnated with guanidine. A beneficial role of guanidine on the reduced copper content has been shown here. In situ TEM has also been carried out in order to evaluate the thermal stability of these nanoparticles and to understand the interest of the functionalization on the sintering of these nanoparticles. Finally, catalytic tests for the selective epoxidation of styrene were also performed on these materials. The preliminary results are very promising for a catalyst preparation process and very simple to implement.

Chapter I: This chapter briefly gives an introduction about microorganisms, their varieties, growth, reproduction etc. In particular, about bacterial function. A sincere attempt is made to review this briefly, including an account of the studies already reported in the literature. Chapter II deals with the antimicrobial activity of Ag/agar film on Escherichia coli (E. coli), Staphylococcus aureus (S. aureus), and Candida albicans (C. albicans). In particular, films were repeatedly cycled for its antimicrobial activity. The antimicrobial activity of Ag/agar film was found to be in the order, C. albicans > E. coli > S. aureus. Chapter III describes the synthesis of Ag@AgI NPs in agarose matrix. A detailed antibacterial studies including repetitive cycles are carried out on E. coli and S. aureus bacteria. EPR and TEM studies are carried out on the Ag@AgI/Agarose and the bacteria, respectively, to elucidate a possible mechanism for killing of the bacteria. The hybrid could be recycled for the antibacterial activity many times and is found to be non toxic towards human cervical cancer cell (HeLa cells). Chapter IV reports the antibacterial efficacy of silver nanoparticles (Ag NPs) deposited alternatively layer by layer (LBL) on chitosan polymer in the form of a thin film over a quartz plate and stainless steel strip against E. coli. AFM studies are carried out on the microbe to know the morphological changes affected by the hybrid film. The hybrid films on aging (3 months) are found to be as bioactive as before. Cytotoxicity experiments indicated good biocompatibility. Chapter V describes the fabrication of carbon foam porous electrode modified with reduced graphene oxide-Ag nanocomposites. The device can perform sterilization by killing pathogenic microbes with the aid of just one 1.5V battery with very little power consumption. Chapter VI Here we have studied in particular a property say the influence of surface defect in the production of ROS by ZnO NPs and in turn the bactericide activity. Secondly, a homogeneous ZnO and ZnO/Ag nanohybrid has been synthesized by employing chitosan as mediator. The synergistic antibacterial effect of ZnO/Ag nanohybrid on bacteria is found to be more effective, compared to the individual components (ZnO and Ag). A possible mechanism has been proposed for the death of bacteria by ZnO/Ag nanohybrid, based on EPR studies and TEM studies.