Dissertation (Ph.D.) -- Auburn University, 2005. / Vita. Includes bibliographical references (ℓ. 117-132).

The purpose of this work was to investigate experimentally the effects of an externally applied magnetic field on free diffusion of saccharides in aqueous solution. The diffusion coefficients of simple saccharides (deoxyribose, D(-)ribose, D(+)xylose, D-glucose, D-galactose, D(-)fructose, lactose, sucrose, maltose, raffinose) diffusing through a 0.4 micron pore diameter Nuclepore membrane were measured in applied magnetic field strengths ranging from zero to 1.1 T. The applied magnetic field strength was the only variable in these experiments. The initial saccharide concentration difference across the membrane was one percent by weight. The experiments were conducted at a constant temperature of 25 ±0.1° C. The diffusion coefficient was obtained by means of a modified Rayleigh interferometer-laser system. A Rayleigh interferometer measured refractive index profiles of dilute saccharide solutions contained in a diffusion cell. Refractive index profiles were converted to concentration profiles which were then used to calculate mass fluxes and the corresponding binary diffusion coefficients. A study of saccharide-water interactions indicates that these interactions are very complex in nature and that saccharide hydration depends not only on the number of equatorial hydroxyl (e-OH) groups in a saccharide molecule but also on their spatial orientation. The saccharide-water solutions exhibit properties that are considered to be the result of two factors (1) the elongated (non-spherical) shape of the oligosaccharides (2) effect of monosaccharides on the local water structure (i.e their ability to either enhance or destroy the local water structure). The observed magnetic field effect on diffusion coefficients of saccharides shows a strong dependency on these two factors. A decrease in binary diffusion coefficients ranging from two to eighteen percent has been observed for applied magnetic fields up to 1.1 T. The diffusion coefficients evaluated at zero field strength (earth's magnetic field) agreed with literature values to within one percent. The noted decrease in diffusivity of monosaccharides (ribose, xylose, galactose, glucose) becomes larger with an increase in the number of equatorial hydroxyl (e-OH number) groups in the saccharide molecule. This is because an increase in e-OH number increases the microviscosity of the saccharide molecule (structure making or stabilising effect). Deoxyribose and fructose, on the other hand, are considered to be structure breakers. The observed decrease in diffusivity for these saccharides induced by the applied magnetic field seem to be the result of a general stabilizing effect of the applied field on the originally less stable saccharide-water solution. The effect of applied magnetic field on the binary diffusion coefficients of oligosaccharides (sucrose, lactose, maltose, raffinose) correlates with the e-OH numbers as well. In this case, however, the observed decrease in diffusivities is due directly to the orientation of these molecules by the externally applied magnetic field (Cotton-Mouton effect). The same membrane was used to study one complete saccharide system, six runs, (made possible by the changes introduced into the design of diffusion cell and diffusion cell holder in this work) so that variation between membranes would not be a factor. The new experimental procedure resulted in significant reduction in data scatter and highly improved measurement accuracy. Finally, it was shown that the membrane only presented an area reduction to diffusion i.e. the transport process through the membrane followed the assumption of free diffusion. / Applied Science, Faculty of / Chemical and Biological Engineering, Department of / Graduate

The objective of the experiments was to compare qualitatively the monosaccharides in the hydrolysis products of the extracellular polysaccharides of several yeasts and yeast-like fungi. Specifically, the study was aimed at finding similarities and differences that might be useful in suggesting and supporting taxonomic relationships. Gas chromatography and paper chromatography were used as methods of analyses in an effort to find out what method is sufficient at the qualitative level for distinguishing some genera of yeasts and yeast-like fungi; and what method would be best at the quantitative level for distinguishing amongst some species of the same genus. From the analytical results it was found that paper chromatography using the solvents ethyl acetate: pyridine: water, (8:2:2) was sufficient for qualitative determination of the monosaccharides in the extracellular polysaccharide hydrolysis products. However, indications were that quantitative analyses by gas chromatography, using the trimethylsilyl derivatives of the monosaccharides would have been successful in distinguishing among species of the same genus. Two groups were formed on the bases of the qualitative results. Group I contained two subgroups. Subgroup I encompassed those yeasts and yeast-like fungi with the monosaccharides galactose, glucose, mannose, xylose present in the hydrolysis products of their extracellular polysaccharides. Included in this Subgroup I are: Cryptococcus laurentii, Tremella mesenterica, Bullera alba, Sporobolomyces odorus, Sporobolomyces singularis, and Rhodotorula glutinis. Subgroup II is Ustilago hordei only, with the monosaccharides galactose, glucose, mannose, and lacking xylose. Group II contains Taphrina populina only, with glucose and mannose present and both galactose and xylose absent. The two groups formed support some of the taxonomic relationships that have already been suggested. The Tremella - Cryptococcus taxonomic relationship that had previously been postulated on the basis of similarities in extracellular polysaccharide hydrolysis products, morphology, carbon assimilation patterns, enzymatic xylosylation reaction, and starch formation was supported. Secondly, the Cryptococcus-Bullera relationship that had been suggested on the basis of inositol assimilation, lack of pseudomycelium, and similarities in starch synthesis, was supported by the qualitative analysis of the monosaccharides present in the extracellular polysaccharide hydrolysis products. The monosaccharides found in both Cryptococcus laurentii and Bullera alba extracellular polysaccharides were the same qualitatively. Duality amongst species of Sporobolomyces might be supported with further work using quantitative gas chromatographic analyses. This duality had been postulated on account of the duality shown in antigenic analyses and percent G+C base analyses of DNA. Taphrina populina can be distinguished from Rhodotorula glutinis and Cryptococcus laurentii. Cryptococcus laurentii produces starch and assimilates inositol: Rhodotorula glutinis assimilates inositol but does not produce starch; and Taphrina populina produces starch but does not assimilate inositol. Two monosaccharides present in the extracellular polysaccharide hydrolysis products of both Cryptococcus laurentii and Rhodotorula glutinis are galactose and xylose whereas Taphrina populina lacks these two monosaccharides. Results obtained from the qualitative analyses of the extracellular polysaccharides produced by fungi may be important taxonomically. This is because the qualitative information may be used when deciding on Perfect-Imperfect fungal relationships. However, this information should be considered along with data from other fields such as morphology, cytology, and genetics before hypothesizing on a taxonomic relationship. / Science, Faculty of / Botany, Department of / Graduate

The specific interaction between saccharide molecules and saccharide-binding proteins, lectins, can be exploited in order to actively target nanostructures to specific cell types for diagnostic or therapeutic purposes. However, the conjugation of saccharides to nanostructures can be complex, leading to a lack of versatility in the functionalities that can be achieved, or poorly characterized and therefore potentially inconsistent. Herein is reported a new procedure by which novel saccharide-containing molecules, for the functionalization of nanostructures, could be synthesized rapidly and with good yields. The synthesis was found to be versatile, providing coating molecules for magnetite nanoparticles and phospholipid vesicles from a number of different hydrazides and reducing sugars. The stability of the synthesized coating molecules was assessed, both in aqueous solution and when bound to the surface of a magnetite nanoparticle, and found to be satisfactory for cell culture purposes. The availability of the saccharide units for lectin binding was confirmed using a Quartz crystal microbalance (QCM-D) in addition to a number of assays. Finally, microscopy techniques were used to study the interactions between saccharide-functionalized magnetite nanoparticles and two cell types, fibroblasts (3T3) and hepatocytes (HepG2).This work demonstrated the ability of saccharide coatings to improve the uptake of nanostructures in a cell culture environment and highlighted the potential cell manipulation application of saccharide-coated magnetite nanoparticles.

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Total polyprenols prepared from Pisum stem and analyzed by HPLC, possessed chain-lengths equivalent to 15-17 isoprene units and were (alpha)-saturated. Pea membranes were supplied with GDP- ('14)C mannose, UDP- ('14)C GlcNAc and UDP- ('14)C glucose as substrates for glycosyltransfer in the presence or absence of added polyprenyl phosphates. Mono- and pyro-phosphorylated fractions were characterized using chromatographic and hydrolytic criteria. Glucosyl and mannosyl transfer took place to form endogenous polyprenyl-P-saccharide. Dolichyl-P was the only added polyprenyl phosphate which acted as an acceptor for these transferases. In contrast, incorporation of all the supplied sugars into polyprenyl-PP-saccharide was stimulated greatly in the presence of relatively short (alpha)-saturated polyprenyl phosphates, e.g., dihydro-heptaprenyl phosphate (7HP). A major product was 7H-PP-monosaccharide. / Oligosaccharide-lipids were also synthesized, all of which could be digested by treatment with endo H, indicating the presence of chitobiose. When UDP- ('14)C GlcNAc was used as substrate in the presence of dolP, labelled dol-PP-chitobiose was formed and then lengthened in the presence of unlabelled GDP-mannose. The same oligosaccharides were formed in experiments where the label was present in mannose. Evidence is presented for the further addition of terminal glucose to form "G" oligosaccharide. Endogenous levels of polyprenyl phosphates clearly limited the activities of pea transglycosylases, which were capable of recognizing isoprenoids of particular chain lengths and saturation patterns.

Saccharides are the main constituents of many Chinese medicinal materials, they include monosaccharides, oligosaccharides and polysaccharides. They have been proved to have many bioactivities such as immunomodulating effects, anti-cancer effects, anti-viral activities, and anti-fatigue effects. Thereby, it is necessary to establish comprehensive qualitative and quantitative analysis of saccharides. On the other hand, saccharides are the main constituent of water extract of many Chinese medicines, which contains a large amount. Up to now, only two botanical drugs, namely Fulyzaq and Veregen, have been approved by the FDA. Despite their complex chemical profiles, 85%-95% of their chemical components could be determined, enabling effective quality control. Therefore, a higher level of qualitative and quantitative analysis is expected for saccharide-rich Chinese medicines. However, currently available methods are weak in terms of accuracy and specificity. Our study aims to develop a comprehensive qualitative and quantitative analysis of saccharides in saccharide-rich Chinese medicines. In this study, a HPLC-NH2P-ELSD quantification method is developed to determine monosaccharides and oligosaccharides. A high performance gel permeation chromatography method was developed to analyze the apparent molecular weight distribution patterns and a 3-Methyl-1-phenyl-2-pyrazoline-5-one derivatization method was used for monosaccharide composition analysis of polysaccharides. The methods were well validated and were then successfully applied in quality assessment of one Chinese medicine material and three Chinese patent drugs from different manufacturers.

Thesis (M.S.)--Youngstown State University, 2008. / Includes bibliographical references (leaves 47-50). Also available via the World Wide Web in PDF format.

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