BACKGROUND:In a globalized word, prevention of infectious diseases is a major challenge. Rapid detection of viable virus particles in water and other environmental samples is essential to public health risk assessment, homeland security and environmental protection. Current virus detection methods, especially assessing viral infectivity, are complex and time-consuming, making point-of-care detection a challenge. Faster, more sensitive, highly specific methods are needed to quantify potentially hazardous viral pathogens and to determine if suspected materials contain viable viral particles. Fourier transform infrared (FTIR) spectroscopy combined with cellular-based sensing, may offer a precise way to detect specific viruses. This approach utilizes infrared light to monitor changes in molecular components of cells by tracking changes in absorbance patterns produced following virus infection. In this work poliovirus (PV1) was used to evaluate the utility of FTIR spectroscopy with cell culture for rapid detection of infective virus particles.RESULTS:Buffalo green monkey kidney (BGMK) cells infected with different virus titers were studied at 1 - 12 hours post-infection (h.p.i.). A partial least squares (PLS) regression method was used to analyze and model cellular responses to different infection titers and times post-infection. The model performs best at 8 h.p.i., resulting in an estimated root mean square error of cross validation (RMSECV) of 17 plaque forming units (PFU)/ml when using low titers of infection of 10 and 100 PFU/ml. Higher titers, from 103 to 106 PFU/ml, could also be reliably detected.CONCLUSIONS:This approach to poliovirus detection and quantification using FTIR spectroscopy and cell culture could potentially be extended to compare biochemical cell responses to infection with different viruses. This virus detection method could feasibly be adapted to an automated scheme for use in areas such as water safety monitoring and medical diagnostics.

A polimerização adequada da resina composta está diretamente relacionada com o sucesso e longevidade das restaurações. Procurando esclarecer a efetividade das fontes de luz na fotoativação, foram realizados estudos laboratoriais, verificando a profundidade de polimerização (1, 2 e 3 mm) da resina composta Filtek Supreme 3M-ESPE, por meio de diferentes análises como: grau de conversão monomérica por espectroscopia com absorção no infravermelho (FT-IR), dureza Vickers, espectroscopia de fluorescência e análise térmica (calorimetria exploratória diferencial - DSC e termogravimetria TG), comparando-se três fontes de luz: lâmpada halógena (Optilux 501-Demetron-Kerr), LED (Lec 1000-MMOptics) e laser de argônio (INNOVA 100-Coherent). Para isso foram confeccionados em triplicata corpos-de-prova, devidamente padronizados para cada condição amostral. Os resultados de dureza Vickers e grau de conversão monomérica por FT-IR receberam tratamento estatístico pela análise de variância (ANOVA) a dois critérios, assim como as comparações múltiplas pelo teste de Tukey ao nível de significância de 5%. Para correlação dos testes de dureza e grau de conversão monomérica foi utilizado o teste de correlação de Pearson (p<0,05). O terceiro milímetro apresentou diferença estatística significante em relação ao primeiro e segundo milímetros tanto nos teste de dureza Vickers, como no grau de conversão monomérica por FT-IR. As fontes de luz apresentaram diferenças estatísticas entre elas na análise do grau de conversão, sendo que a lâmpada halógena obteve os melhores resultados, seguido do Led e laser de argônio. Não houve uma correlação positiva entre os testes de dureza e grau de conversão por FT-IR. Nos testes de espectroscopia de fluorescência e análise térmica foi possível observar uma mesma tendência de resultados verificado no teste de grau de conversão por FT-IR em relação às diferenças entre as fontes de luz e as profundidades de polimerização. Os testes mostraram-se efetivos na avaliação da profundidade de polimerização, porém o teste de dureza não se correlacionou com os demais testes e todas as fontes de luz se mostraram aptas a polimerização da resina composta até 2 mm de profundidade de acordo com os parâmetros utilizados nessa pesquisa. / The adequate polymerization of composite resin is directly related with the restoration success and longevity. Experimental studies were made to verify the efficiency of light sources in the photoactivation by the evaluation of the depth of cure (1, 2 and 3 mm) of composite resin Filtek Supreme 3M-ESPE by several methods: degree of monomeric conversion by spectroscopy Fourier transform infrared (FT-IR), Vickers hardness, fluorescence spectroscopy and thermal analysis (differential scanning calorimetry - DSC and termogravimetric analysis - TGA), comparing 3 light sources: halogen light (Optilux 501/Demetron-Kerr), LED (Lec 1000-MMOptics) and argon laser (INNOVA 100-Coherent). Samples were made in triplicate and standardized for each condition. The results of Vickers hardness and degree of conversion by FT-IR were subjected to statistical analysis using two-way analysis of variance (ANOVA) and Tukey test for multiple comparisons and the statistical significance was assumed at 5% (p<0.05), and for correlation the tests of Vickers hardness and degree of conversion were used the Pearson\' s test (p<0.05). The third millimeter presented statistical difference in relation with first and second millimeters for both tests: Vickers hardness and degree of conversion by FT-IR. The light sources presented statistical differences among them in the degree of conversion test: the halogen light had better values followed by LED and argon laser. And did not have a positive correlation between Vickers hardness and degree of conversion FT-IR. In the fluorescence spectroscopy and thermal analysis was possible to observe the same tendency in the results that they had in the degree of conversion test, in relation of difference in depth of cure and light sources. All tests showed strength in the evaluation of depth of cure, however the Vickers hardness test were not correlated with other tests and all light sources showed to able to polymerized the composite resin until 2 millimeters according to the parameters used in this research.

A polimerização adequada da resina composta está diretamente relacionada com o sucesso e longevidade das restaurações. Procurando esclarecer a efetividade das fontes de luz na fotoativação, foram realizados estudos laboratoriais, verificando a profundidade de polimerização (1, 2 e 3 mm) da resina composta Filtek Supreme 3M-ESPE, por meio de diferentes análises como: grau de conversão monomérica por espectroscopia com absorção no infravermelho (FT-IR), dureza Vickers, espectroscopia de fluorescência e análise térmica (calorimetria exploratória diferencial - DSC e termogravimetria TG), comparando-se três fontes de luz: lâmpada halógena (Optilux 501-Demetron-Kerr), LED (Lec 1000-MMOptics) e laser de argônio (INNOVA 100-Coherent). Para isso foram confeccionados em triplicata corpos-de-prova, devidamente padronizados para cada condição amostral. Os resultados de dureza Vickers e grau de conversão monomérica por FT-IR receberam tratamento estatístico pela análise de variância (ANOVA) a dois critérios, assim como as comparações múltiplas pelo teste de Tukey ao nível de significância de 5%. Para correlação dos testes de dureza e grau de conversão monomérica foi utilizado o teste de correlação de Pearson (p<0,05). O terceiro milímetro apresentou diferença estatística significante em relação ao primeiro e segundo milímetros tanto nos teste de dureza Vickers, como no grau de conversão monomérica por FT-IR. As fontes de luz apresentaram diferenças estatísticas entre elas na análise do grau de conversão, sendo que a lâmpada halógena obteve os melhores resultados, seguido do Led e laser de argônio. Não houve uma correlação positiva entre os testes de dureza e grau de conversão por FT-IR. Nos testes de espectroscopia de fluorescência e análise térmica foi possível observar uma mesma tendência de resultados verificado no teste de grau de conversão por FT-IR em relação às diferenças entre as fontes de luz e as profundidades de polimerização. Os testes mostraram-se efetivos na avaliação da profundidade de polimerização, porém o teste de dureza não se correlacionou com os demais testes e todas as fontes de luz se mostraram aptas a polimerização da resina composta até 2 mm de profundidade de acordo com os parâmetros utilizados nessa pesquisa. / The adequate polymerization of composite resin is directly related with the restoration success and longevity. Experimental studies were made to verify the efficiency of light sources in the photoactivation by the evaluation of the depth of cure (1, 2 and 3 mm) of composite resin Filtek Supreme 3M-ESPE by several methods: degree of monomeric conversion by spectroscopy Fourier transform infrared (FT-IR), Vickers hardness, fluorescence spectroscopy and thermal analysis (differential scanning calorimetry - DSC and termogravimetric analysis - TGA), comparing 3 light sources: halogen light (Optilux 501/Demetron-Kerr), LED (Lec 1000-MMOptics) and argon laser (INNOVA 100-Coherent). Samples were made in triplicate and standardized for each condition. The results of Vickers hardness and degree of conversion by FT-IR were subjected to statistical analysis using two-way analysis of variance (ANOVA) and Tukey test for multiple comparisons and the statistical significance was assumed at 5% (p<0.05), and for correlation the tests of Vickers hardness and degree of conversion were used the Pearson\' s test (p<0.05). The third millimeter presented statistical difference in relation with first and second millimeters for both tests: Vickers hardness and degree of conversion by FT-IR. The light sources presented statistical differences among them in the degree of conversion test: the halogen light had better values followed by LED and argon laser. And did not have a positive correlation between Vickers hardness and degree of conversion FT-IR. In the fluorescence spectroscopy and thermal analysis was possible to observe the same tendency in the results that they had in the degree of conversion test, in relation of difference in depth of cure and light sources. All tests showed strength in the evaluation of depth of cure, however the Vickers hardness test were not correlated with other tests and all light sources showed to able to polymerized the composite resin until 2 millimeters according to the parameters used in this research.

The feasibility of FTIR-based spectroscopy as a tool to measure cellular response to therapeutics was investigated. Fourier transform mid-infrared spectroscopy has been used in conjunction with multivariate analysis (MVA) to assess the chemistry of many clinically relevant biological materials; however, the technique has not yet found its place in a clinical setting. One issue that has held the technique back is due to the spectral distortions caused by resonant Mie scattering (RMieS), which affects the ability to confidently assign molecular assignments to the spectral signals from biomaterials. In the light of recently improved understanding of RMieS, resulting in a novel correction algorithm, the analytical robustness of corrected FTIR spectra was validated against multi-discipline methods to characterise a set of renal cell lines which were selected for their difference in morphology.After validation of the FTIR methodology by discriminating different cell lines, the second stage of analyses tested the sensitivity of FTIR technique by determining if discrete chemical differences could be highlighted within a cell population of the same origin. The renal carcinoma cell line 2245R contains a sub-population to contain a sub-population of cells displaying 'stem-cell like' properties. These stem-like cells, however, are difficult to isolate and characterise by conventional '-omic' means. Finally, cellular response to chemotherapeutics was investigated using the established renal cell lines CAKI-2 and A-498. For the model, 5-fluorouracil (5FU), an established chemotherapeutic agent with known mechanisms of action was used. Novel gold-based therapeutic compounds were also assessed in parallel to determine their efficacy against renal cell carcinoma. The novel compounds displayed initial activity, as the FTIR evidence suggested compounds were able to enter the cells in the first instance, evoking a cellular response. The long-term performance, tracked with standard proliferation assays and FTIR spectroscopy in the renal cancer cell model, however, was poor. Rather than dismissing the compounds as in-active, the compounds may simply be more effective in cancer cell types of a different nature. The FTIR-based evidence provided the means to suggest such a conclusion. Overall, the initial results suggest that the combination of FTIR and MVA, in the presence of the novel RMieS-EMSC algorithm can detect differences in cellular response to chemotherapeutics. The results were also in-line with complimentary biological-based techniques, demonstrating the powerful potential of the technique as a promising drug screening tool.

M. Tech. (Department of Chemical Engineering, Faculty of Engineering and Technology), Vaal University of Technology. / The industrialisation of the twenty-first century and the worldwide population growth led to the high demand for energy. Fossil fuels are the leading contributor to the global energy, and subsequently, there is a high demand of fuels. The decrease of global fossil fuels and the environmental air pollution caused by these fuels are concerning. Therefore, eco-friendly and renewable fuel such as biodiesel is one the leading alternative. Chromatography and Spectroscopy are the most used analytical methods and proven reliable but are time-consuming, requires qualified personal, extensive samples preparation, costly and do not provide in-situ real-time monitoring. Fourier Transform Infrared (FTIR) has mainly been used for qualitative analysis of biodiesel, but not much work has been reported in real-time monitoring. This study focused on the modelling of in-situ real-time monitoring of the biodiesel production from sunflower oil using FTIR (Fourier Transform Infrared). The first part of the study investigated the effect of catalyst ratio and methanol to oil ratio on biodiesel production by using central composite design (CCD). Biodiesel was produced by transesterification using Sodium Hydroxide as a homogeneous catalyst. A laboratory-scale reactor consisting of; flat bottom flask mounted with a reflux condenser, a hot plate as heating element equipped with temperature, timer and stirring rate regulator was used. Key parameters including, time, temperature and mixing rate, were kept constant at 60 minutes, 60 oC and 600 RPM, respectively. From the results obtained, it was observed that the biodiesel yield depends on catalyst ratio and methanol to oil ratio. The highest yield of 50.65 % was obtained at a catalyst ratio of 0.5 wt% and methanol to oil mole ratio 10.5. The analysis of variances of biodiesel yield showed the R2 value of 0.8387. A quadratic mathematical model was developed to predict the biodiesel yield in the specified parameters range. The same set-up was used to produce waste margarine biodiesel using a homogeneous catalyst, potassium hydroxide (KOH). The effects of four reaction parameters were studied, these were: methanol to oil ratio (3:1 to 15:1), catalyst ratio (0.3 to 1.5 wt. %), temperature (30 to 70 oC), time (20 to 80 minutes). The highest yield of 91.13 % was obtained at 60°C reaction temperature, 9:1 methanol to oil molar ratio, 0.9 wt. % catalyst ratio and 60 minutes. The important biodiesel fuel properties were found to be within specifications of the American Standard Test Method specifications (ASTM). It was concluded that waste margarine can be used to produce biodiesel as a low-cost feedstock. The core of the study was performed using EasyMax Mettler Toledo reactor equipped with a DiComp (Diamond) probe. The quantitative monitoring of the biodiesel production was performed by building a quantitative model with multivariate calibration using iC Quant module from iC IR 7.0 software. Fourteen samples of known concentrations were used for the modelling which were taken in duplicate for model calibration and cross-validation, data were pre-processed using mean centring and variance scale, spectrum math square root and solvent subtraction. These pre-processing methods improved the performance indexes from 7.98 to 0.0096, 11.2 to 3.41, 6.32 to 2.72, 0.9416 to 0.9999, RMSEC, RMSECV, RMSEP and R2Cum, respectively. The R2 values of 1 (training), 0.9918 (test), 0.9946 (cross-validation) indicated the fitness of the model built. The model was tested against the univariate model; small discrepancies were observed at low concentration due to unmodelled intermediates but were quite close at concentrations above 18%. The software eliminated the complexity of the Partial Least Square (PLS) chemometrics. It was concluded that the model obtained could be used to monitor transesterification of sunflower oil at industrial and lab scale. The model thus obtained, a batch reactor setup, EasyMax Mettler Toledodo reactor was used, the experiments were designed and monitored using iControl software. The results were recorded and quantified using iC IR software based on the biodiesel calibrated monitoring model built. The optimisation of the biodiesel was performed using three key parameters (methanol to oil ratio, catalyst ratio and temperature) while keeping time at 60 minutes and mixing rate at 150RPM. The highest yield of 97.85 % was obtained at 60 oC, 0.85 wt % catalyst ratio and 10.5 methanol to oil mole ratio. The analysis of variances of biodiesel production showed the values of 0.9847, 0.9674 and 0.8749, for R-squared, adjusted R-squared and predicted R-squared, respectively. A quadratic mathematical model was developed to predict the biodiesel conversion in the specified parameters ranges. Using the Arrhenius equation, activation energy (Ea) and frequency factor were found to be 41.279 kJ.mole-1 and 1.08 x10-4 M-1. s-1, respectively. The proposed kinetics model was a pseudo-first-order reaction. It was concluded that the model obtained can be used for industrial and laboratory-scale biodiesel production monitoring.

Disease- and drug-related bone disorders are rapidly increasing in the population. The drugs which are used for the treatment of neurodegenerative diseases and metabolic derangements, may have negative or positive effects on bone tissues. In the first study, the possible side-effects of Carbamazepine and epileptic seizures on bone structure and composition were investigated by FTIR and synchrotron-FTIR microspectroscopy, AFM and micro- and nano-hardness analysis. The effects on the blood parameters, bone turnover and vitamin D metabolism were also investigated by ELISA and western blot analysis. The current study provides the first report on differentiation of the effects of both epileptic seizures and AED therapy on bones. Besides Carbamazepine treatment, seizures also caused a decrease in the strength of bone. The biochemical data showed that both the epileptic and drug-treated groups decreased vitamin D levels by increasing the vitamin D catabolism enzyme / 25-hydroxyvitamin D-24-hydroxylase. In the second study, the possible pleiotropic (positive) effects of cholesterol lowering drug / Simvastatin on bones were investigated by ATR-FTIR spectroscopy. The current study provides the first report on dose-dependent effects of simvastatin on protein structure and lipid conformation of bones. ATR-FTIR studies showed that although both high and low dose simvastatin strengthen bones, low dose simvastatin treatment is much more effective in increasing bone strength. Neural network analysis revealed an increased antiparallel and aggregated beta sheet and random coil in the protein secondary structure of high dose group implying a protein denaturation. Moreover, high dose may induce lipid peroxidation which limit the pleiotropic effects of high dose treatment on bones. This study clearly demonstrated that using low dose simvastatin is safer and more effective for bone health than high dose simvastatin treatment.

Bone marrow mesenchymal stem cells (BM-MSCs) are pluripotent cells that can differentiate into a variety of non-hematopoietic tissues. They also maintain healthy heamatopoiesis by providing supportive cellular microenvironment into BM. In this thesis, MSCs were characterized in terms of their morphological, immunophenotypical and differentiation properties. Then, they were examined by attenuated total reflection-Fourier transform infrared (ATR-FTIR) spectroscopy together with hierarchical clustering, and FTIR microspectroscopy. In the first part of this study, global structural and compositional changes in BM-MSCs during beta thallasemia major (

This dissertation presents a study of the molecular mechanism underlying the highly efficient solar energy conversion processes that occur in the Photosystem I (PS I) reaction centers in plants and bacteria. The primary electron donor P700 is at the heart of solar energy conversion process in PS I and the aim is to obtain a better understanding of the electronic and structural organization of P700 in the ground and excited states. Static Fourier Transform Infra-Red (FTIR) difference spectroscopy (DS) in combination with site directed mutagenesis and Density Functional Theory (DFT) based vibrational frequency simulations were used to investigate how protein interactions such as histidine ligation and hydrogen bonding modulate this organization. (P700+-P700) FTIR DS at 77K were obtained from a series of mutants from the cyanobacterium Synechocystis sp. 6803 (S. 6803) where the amino acid residues near the C=O groups of the two chlorophylls of P700 where specifically changed. (P700+-P700) FTIR DS was also obtained for a set of mutants from C. reinhardtii where the axial ligand to A0-, the primary electron acceptor in PS I was modified. The FTIR DS obtained from these mutants provides information on the axial ligands, the hydrogen bonding status as well as the polarity of the environment of specific functional groups that are part of the chlorophyll molecules that constitute P700. Assignment of the FTIR bands to vibrational modes in specific types of environment is very difficult. In order to assist the assignment of the difference bands in experimental spectra DFT based vibrational mode frequency calculations were undertaken for Chl-a and Chl-a+ model molecular systems under different set of conditions; in the gas phase, in solvents using the Polarizable Continuum Model (PCM), in the presence of explicit solvent molecules using QM/MM methods, and in the presence of axial ligands and hydrogen bonds. DFT methods were also used to calculate the charge, spin and redox properties of Chl-a/Chl-a’ dimer models that are representative of P700, the primary electron donor in PS I.

Bone marrow mesenchymal stem cells (BM-MSCs) are pluripotent cells that can differentiate into a variety of non-hematopoietic tissues. They also maintain healthy heamatopoiesis by providing supportive cellular microenvironment into BM. In this thesis, MSCs were characterized in terms of their morphological, immunophenotypical and differentiation properties. Then, they were examined by attenuated total reflection-Fourier transform infrared (ATR-FTIR) spectroscopy together with hierarchical clustering, and FTIR microspectroscopy. In the first part of this study, global structural and compositional changes in BM-MSCs during beta thallasemia major (

High performance multifunctional epoxy resin systems are becoming increasingly important as matrix materials for the advanced composites used in aerospace, electronics, automotive and other industries. In a composite based on epoxy resin systems, a three-dimensional network of the matrix is formed around the reinforcing fibre as a result of the chemical reaction between the resin and the curing agent. This chemical process, known as curing, is an important event to he considered in the production of composite components made up of these resin systems. Two process parameters namely viscosity and chemical conversion are of paramount significance in the production of composite materials Curing studies of the resin systems based on these two parameters, would therefore assume great importance in deciding the performance reliability of the end product. The objectives of the present investigation are 1. to study the cure kinetics of three thermoset resin systems, viz., i) epoxy novolac (EPIT)/ diamino diphenyl methane{DDM), ii) trigylcidyl para- ammo phenol (TGPAP)/toluene diamine (TDA) and iii) tetraglycidyl diamino diphenyl methane (TGDDM)/pyridine diamine(PDA) using the cure kinetic models based on chemical conversion (α), Theological conversion (β) and viscosity. 2.to develop a correlation between a and viscosity (η) and modify an existing autocatalytic model based on α, to the viscosity domain and 3.to investigate the cure behaviour of these systems in terms of the TTT cure diagram and its associated models. EPN/DDM, TGPAP/PDA and TGDDM/PDA resin systems were chosen for the studies to represent a range of functionalities, The cure was monitored using differential scanning calorimetry (DSC), fourier transform infrared (FTIR) and dynamic mechanical analysis (DMA) techniques by following the changes in enthalpy, functional groups and rheology, respectively. The kinetic parameters namely, order of reaction and activation energy were estimated from dynamic DSC data using the methods of Freeman-Carroll and Ellerstein using nth rate expression. Barton, Kissinger and Osawa methods were employed to find out the activation energy from the peak/equal conversion at different heating rates. Isothermal DSC data were also analyzed using nth order model and it was observed that the data could be fitted satisfactorily only for higher temperatures The results obtained from the analysis of both dynamic and isothermal DSC data using nth order model clearly indicate that this model is inadequate for describing the cure behavior. The isothermal DSC data was analyzed by the autocatalytic models of Hone and Kamal Good correlation was observed with Hum and Kamal models up to 60-70%, 25% and 45% conversions for EPN/DDM, TGPAP/TDA and TCDDM/PDA systems respectively. However, the parameters m and n in Kamal model were found to be temperature dependent for EPN/DDM and TCPAP/TDA systems. The limited applicability of the autocatalytic models IK attributed to the counter-effect offered by the intra-molecular bonding taking place. The primary amine and epoxy groups conversions obtained from FTIR were analyzed using autocatalytic model and the kinetic parameters were calculated. The reactivity ratio of the primary amine and the secondary amine with epoxy was found to be dependent on temperature in agreement with the recent findings reported m the literature. The existing models that relate the cure kinetics and the rheological changes, are dual Arrhenius nth order model and autocatalytic model The nth order kinetic model was used to evaluate the kinetic parameters using the viscosity data at different cure temperatures under isothermal conditions As the storage modulus, G' is proportional to the chemical cross links and becomes significant only after the g<4 point, it was used to follow the changes in conversion known as rheoconversion after the gel point The rheoconversion was found by normalizing the G' data with G1^, the storage modulus of the fully cured resin It was used to study the cuie kinetics using an autocatalytic model The kinetic parameters such as rate constant, acceptation and retardation parameters were evaluated and that temperature dependence was established. While the existing models relate viscosity and conversion only up to gel point the new proposed model, termed VISCON model takes into account the changes up to vitrification. The relation so developed is used to modify the autocatalytic cure model based on chemical conversion. The parameters appearing in this model were evaluated using Levenberg-Marquardt error minimization algorithm. The kinetic parameters obtained are comparable with the values estimated using the DSC data. All the models cited above represent the microkinetic aspects. The models based on the information of TTT cure diagrams, however, represent the macrokinetic aspects of the cure, as they are based on the cure stages such as gelation and vitrification TTT diagram relates the cure characteristics like cure temperature, cure time, Ta and, indirectly, chemical conversion Hence the ultimate properties of the composite could he predicted and established with the help of the models based on TTT cure diagrams The changes in the storage modulus, G1 and loss modulus, G", were followed to identify the gel and vitrification points of the resin systems at different cure temperatures Gel point and vitrification point were used to generate gelation and vitrification hues in the construction of TTT cure diagrams for EPN/DDM, TGPAP/TDA and TGDDM/PDA resin systems Theoretical TTT diagrams were generated and IBO-T, contours were established using the TTT diagram-based models The cure schedule for the resin systems investigated could be determined from the TTT diagram and the respective rheological data.