NMR spectroscopy is a very powerful technique for the characterization of structure and dynamics of a variety of systems starting from small organic molecules to large biological macromolecules. In solids, the study of protons becomes more interesting because they are very sensitive to inter-molecular packing and are directly involved in hydrogen-bonding and aromatic π-π interactions, etc. The present thesis is devoted essentially to utilizing information from proton resonances obtained using multinuclear and multiple-quantum approaches. The thesis has two parts. The first part deals with methodological developments in the area of solid-state NMR, relevant to the study of rigid powder samples as well as partially ordered liquid crystalline materials. Methods have been proposed to investigate the structure of small molecules at moderate spinning frequencies and thermotropic liquid crystals at static conditions. Proton detected heteronuclear experimental methods based on both first and second-order cross polarization at moderate and ultra-fast magic angle spinning rates are also proposed. The second part of the thesis deals with the application of both newly proposed and existing solid state NMR methods to the study of several biologically relevant systems. These include the study of several designed as well as naturally occurring peptides. The use of first-principles calculations based on GIPAW method for supporting the experimentally obtained results has also been made. The thesis is divided into five chapters. In the second chapter, a new pulse sequence to correlate Double Quantum (DQ) proton frequencies to carbon Single Quantum (SQ) chemical shifts in the solid state has been proposed. In this sequence, named as MAS-J-1H (DQ)-13C-HMQC, the correlation between 1H and 13C is achieved through scalar coupling, while the double-quantum coherence among protons is generated through dipolar couplings. This experiment is particularly suited for the study of 13C in natural abundance. The advantages of the technique with applications to alanine, histidine and a model liquid crystalline material have been demonstrated. The assignment of 13C spectra of partially ordered systems has also been considered. In this case the assignment of the spectrum is a major challenge due to the interplay of anisotropic order and chemical shift parameters. The DQ-SQ correlation experiment described in the thesis has been applied to a well known liquid crystal and also to a novel thiophene based liquid crystal and the local order parameters of the liquid-crystal have been obtained. The thesis also presents results on the azelaic acid -isonicotinamide co-crystal as well as the drug ibuprofen obtained by using novel methodologies. In the case of the former, the problem of overlap of resonances was overcome with the use of the REVERSE-CP approach to separate out the carbon attached protons from the rest of the protons. Subsequently, by the use of several combined approaches, the structural features were identified. A new heteronuclear correlation pulse sequence for solids under fast MAS conditions has also been tested. With low r.f powers, a second-order dipolar term mediated transfer of magnetization between I and S spin known as second order cross-polarization (SOCP) was exploited to obtain the entire spin system connectivity. Both carbon detected and proton detected experiments have been carried out and their utility evaluated. Similar approaches to shed light on the structure and conformation of a set of proline and pseudoproline based designed β-turn peptides that are used as templates for understanding protein folding have been made. Results of studies on two biologically important forms of the short-chain peptides namely glutathione reduced (GSH) and oxidized (GSSG) tripeptides are also presented.

L'objet de cette thèse est l'étude de l'interaction d'un laser femtoseconde avec un support photosensible particulier: un verre phosphate dopé à l'argent appelé verre photo-luminescent (PL). Une nouvelle approche permettant de réaliser en trois dimensions dans un verre PL des nanostructures d'argent aux dimensions bien inférieures à la limite de diffraction est tout d'abord présentée. La mesure des propriétés optiques et structurales pour différentes échelles (spatiales et temporelles) a permis de proposer un mécanisme de formation des structures photo-induites qui est basé sur un jeu subtil entre les phénomènes d’absorption non-linéaire et de thermo-diffusion. La deuxième partie de cette thèse sera orientée sur les propriétés optiques (linéaires et non-linéaires) et les applications des ces nanostructures d’argent. En particulier, l’exaltation des propriétés non-linéaires des agrégats d’argent sera exploitée pour stocker optiquement de l’information en trois dimentions. / The aim of this work is the study of the interaction between a femtosecond laser and a special photosensitive medium: a silver containing phosphate glass, also called photo-luminescent (PL) glass. A new approach allowing to write, inside the PL glass, 3D silver nanostructures with feature size down to the diffraction limit is presented. Based on optical and structural measurments at different time and spacial scales, the mechanism of the formation of these nanostructures is described. A subtle interplay between nonlinear absorption and thermo-diffusion effects is found to be the key of the mechanism. The second part of this work relies on the optical properties (linear and nonlinear) and few applications of the silver nanostructures. More particulary, the enhancement of their nonlinear properties is used for three dimentional optical data storage.

Exogenously supplied organoselenium compounds, capable of propagating a selenium redox cycle, were shown to supplement natural cellular defenses against oxidants generated during biological activity. Phenylaminoalkyl selenides were developed in our laboratory as novel substrate analogs for the enzyme dopamine beta-monooxygenase. Recently, phenylaminoalkyl selenides were found to protect plasmid DNA and Molecular beacons from oxoperoxynitrate – mediated damage by scavenging this oxidant and forming the corresponding selenoxides as the sole selenium – containing products. Rate constants were determined for the reactions of the phenylaminoalkyl selenoxides with GSH at physiological pH and 25 degrees C. The kinetic data obtained in current and previous research was subsequently used in a MatLab simulation, which showed the feasibility of selenium redox cycling by GSH in the presence of a cellular oxidant, oxoperoxynitrate. Loblolly pine (LP, Pinus taeda) is the primary commercial species in southern forests covering 11.7 million hectares. Somatic embryogenesis (SE) is an effective technique to implement production of high value genotypes of LP. SE is a multi-step process, which includes initiation of somatic embryo (SME) growth from tree tissue, maintenance and multiplication of early stage SMEs and the maturation / germination phase. In this work, we isolated a substance from stage 2 or 3 LP female gametophyte (FG) tissue that stimulates early stage SME growth, and characterized this substance as citric acid on the basis of 1H NMR and mass spectrometry. We then demonstrated that topical application of citric acid to SMEs stimulates embryo colony growth at p = 0.05 for 3 of the 5 genotypes tested. Phenylketonuria (PKU) is an autosomal recessive disorder caused by an impaired conversion of L-phenylalanine (L-Phe) to L-tyrosine (L-Tyr). A novel assay based on enzymatic - colorimetric methodology (ECA) was developed in order to detect elevated concentrations of L-Phe in undeproteinized plasma of PKU patients via continuous spectrophotometric detection. We report here that L-Phe concentrations in undeproteinized plasma measured using our ECA were comparable to those determined on an amino acid analyzer based on Pearson correlation coefficients and a Bland and Altman comparison.

Liver disease is now a leading cause of non-AIDS related morbidity and mortality in people living with HIV (PLWH). The present study investigated the interplay between adverse lifestyle factors that are prevalent in PLWH, biological mediators of liver pathogenesis, and a non-invasive measure of liver fibrosis (FIB-4 index) in HIV mono- and HIV/HCV co-infected individuals. The results of this investigation in the Miami Adult Studies of HIV (MASH) cohort show that the odds of liver fibrosis progression significantly increased over two years for HIV mono-infected participants who drank alcohol hazardously (OR 3.038, P=0.048), and had BMI ≥ 28kg/m2 (OR 2.934, P=0.027). Cocaine use reduced the odds of advancing one stage of liver fibrosis (OR 0.228, P=0.038), but an interaction between high BMI and cocaine use slightly raised the odds by 4.8% of liver fibrosis progression (P=0.072). HIV/HCV co-infected participants showed interactions between cocaine use and high BMI with increased FIB-4 stage (OR 4.985, P= 0.034), however no lifestyle factors could independently predict FIB-4 stage in this group. Biological mediators previously associated with liver pathogenesis were associated with higher FIB-4 index over 2 years in a subset of (n=65) HIV mono-infected participants. Plasma measures of oxidative stress (% oxidized glutathione: OR 4.342, P= 0.046), hepatocyte-specific apoptosis (Cytokeratin-18 (CK-18): OR 1.008, P=0.021), and microbial endotoxin (lipopolysaccharide (LPS): OR 1.098, P= 0.097) were associated with having higher odds of progressing at least one stage of FIB-4 over 2 years. The same biological mediators were also associated with liver fibrosis within HIV infected people who also had a harmful lifestyle characteristic. FIB-4 index was significantly associated with % oxidized glutathione in obese subjects (β=0.563, P=0.018), TGF-β1 in cocaine users (β=0.858, P=0.027), and CK-18 in HIV infected individuals without any adverse lifestyle factors (β=0.435, P=0.015). Taken together, the findings of these studies describe interrelationships between HIV disease status, lifestyle, and biological mediators of liver fibrosis. The results show interactions between lifestyle conditions and the mediators of liver fibrosis may account for higher rates of liver disease in HIV infection. Research is warranted to develop personalized therapeutics for PLWH to curb the burden of liver disease.