There has been increasing interest in the syntheses of carbohydrates due to their profound role in various biological processes and thus their potential in drug development. Various methods have been developed for the preparation of these oligosaccharides from simpler carbohydrate derivatives both chemically and enzymatically but they must be carefully applied to obtain the desired linkages. The main method that has been used to control the regioselectivity of glycosylations is protecting group chemistry. Unfortunately this requires additional steps lengthening the synthesis sequence. Extensive literature has shown the ability for carbohydrate recognition via organoboron methods through selective binding and their ability to undergo regioselective glycosylation through sugar-derived boronate esters. Exploiting these factors and in extension of previous work done in our laboratory on borinic acid- and boronic acid-catalyzed regioselective glycosylations of carbohydrate derivatives, we proposed a target oriented synthesis of a pentassacharide moiety found in four saponins isolated from Spergularia ramosa.

The synthesis of glycoconjugates is of interest in biological and medicinal research. There are numerous approaches to the synthesis of glycosides involving protecting group free methods. This thesis outlines what has been achieved in the field and two novel approaches for O-glycosidation. The first approach involves the use of a toluenesulfonohydrazide glycoside with a purification handle designed for simple glycoside purifications. The butyl 3-O-octyl-D-glucopryanoside was successfully synthesized but did not have the desired property of yielding simple-to-purify glycosides as products. The second approach uses a thiouronium glycosyl donor; a variety of glycosidations using this donor were investigated. The glucosyl thiouronium salt donor was shown to undergo glycosidation effectively with simple alcohols.

There has been increasing interest in the syntheses of carbohydrates due to their profound role in various biological processes and thus their potential in drug development. Various methods have been developed for the preparation of these oligosaccharides from simpler carbohydrate derivatives both chemically and enzymatically but they must be carefully applied to obtain the desired linkages. The main method that has been used to control the regioselectivity of glycosylations is protecting group chemistry. Unfortunately this requires additional steps lengthening the synthesis sequence. Extensive literature has shown the ability for carbohydrate recognition via organoboron methods through selective binding and their ability to undergo regioselective glycosylation through sugar-derived boronate esters. Exploiting these factors and in extension of previous work done in our laboratory on borinic acid- and boronic acid-catalyzed regioselective glycosylations of carbohydrate derivatives, we proposed a target oriented synthesis of a pentassacharide moiety found in four saponins isolated from Spergularia ramosa.

With the rapidly increasing interest in the field of glycomics, which is the comprehensive study of the roles carbohydrates play in a living system, urgent need for developing quick and highly selective carbohydrate sensors is growing. The boronic acid group, with its electron-deficient structure (6 valence electrons with an open shell), acts as a Lewis acid with high intrinsic affinity towards Lewis bases such as fluoride, cyanide and hydroxyl groups. Specifically, formation of a 5- or 6- membered ring between the boronic acid moiety and a1,2- or 1,3-diol in aqueous solution has been fully explored as a strategy of carbohydrate sensor design. Along this line, those binders were termed as ¡°boronolectins¡± because of their similar functions as lectins. One challenge in developing boronic acid-based carbohydrate sensors is to enhance the discriminating ability among various carbohydrate analytes with diverse building blocks and complex linkage patterns. One approach is using polypeptide or oligonucleotide as a backbone or scaffold with functionalized boronic acid moiety to create a molecular library, and then selecting binders with favorable properties. The work presented here includes three general research parts: synthesis of a naphthalimide-based boronic acid-conjugated thymidine triphosphate (NB-TTP), fluorescence property studies of NB-TTP incorporated DNA (NB-DNA), and cellular imaging studies using NB-TTP: 1) 4-Amino-1,4-naphthalimide (Nap) was chosen as the fluorophore because of its relatively long excitation and emission wavelengths, and stability. The synthesis of naphthalimide-based boronic acid (NB) followed similar route as previously published work. Tethering of boronic acid moiety and TTP was accomplished through Cu(I)-catalyzed azide-alkyne cyclization (CuAAC), known as click chemistry. The synthesized NB-TTP showed fluorescence enhancements at long wavelength (¦Ëem: 540 nm) upon sugar addition. 2) NB-TTP was incorporated into DNA through Klenow fragment-catalyzed primer extension reactions. Different DNA sequences were designed with varying number and spacing for NB-TTP incorporation. The preliminary study provided certain insight into several factors that affect the fluorescent properties of different NB -DNA. 3) NB-TTP was added into Hela cell culture medium to study its cell imaging properties. With the observation under fluorescent microscope, it was demonstrated that NB-TTP showed good cell membrane permeability and significant accumulation in cell nucleus.

The glycemic index (GI) of a pre-exercise meal has been shown to affect substrate oxidation during exercise and may influence exercise performance. Previous research in this area has focused on continuous, moderate intensity exercise. The purpose of this study was to examine the effects of low and high glycemic index (GI) pre-exercise meals on metabolism and performance during soccer-specific intermittent exercise. Thirteen trained male soccer players (22.3 ± 3.3 yrs) participated in four experimental trials in a repeated crossover design. Isocaloric low GI-high protein (lentils), high GI-high protein (potato + egg whites), or high GI-low protein (potato) meals were consumed two hours before a 90-minute treadmill soccer match simulation. A fasted control condition was also employed. Blood and expired gas samples were collected before and during exercise to assess markers of carbohydrate and fat metabolism. The distance covered on five 1-minute sprints (separated by 2.5 minutes of recovery) performed during the last 15 minutes of the match was used to assess performance. Serum insulin concentration at the start of exercise was higher in the high GI-low protein condition compared to all other conditions (p<0.001). During exercise, the rate of carbohydrate oxidation was significantly higher and the rate of fat oxidation was significantly lower in the high GI-low protein condition compared to control (p<0.05). The distance covered on sprints 1 and 2 was significantly greater in the low GI-high protein condition compared to control (p<0.05). The distance covered on sprint 2 was significantly greater in the high GI-high protein condition compared to control (p<0.05). There were no significant differences between conditions for sprints 3-5. Ratings of perceived exertion (RPE) throughout exercise were significantly lower in the low GI-high protein condition compared to both control and high GI-low protein conditions (p<0.05). It is concluded that low GI-high protein and high GI-high protein pre-exercise meals result in small improvements in initial repeated sprint performance compared to fasted control. Performance was not improved following a high GI-low protein pre-exercise meal compared to control, indicating that the increased carbohydrate oxidation and decreased fat oxidation in this condition may have negated any potential performance advantage of carbohydrate consumption. The lower RPE throughout exercise in low GI-high protein compared to control and high GI-low protein indicates a possible beneficial effect of consuming a low GI meal prior to soccer-specific intermittent exercise.

Bacterial biofilms are surface associated colonies that are of considerable concern and interest to industry, medicine and research. They are resistant to antibiotics, their host’s defences and are able to survive under harsh conditions. Biofilm formation in many bacterial strains are dependent on the production of a polysaccharide intercellular adhesion (PIA), a beta-(1-->6)-N-acetylglucosamine polymer. Vaccines derived from biologically isolated PIA have shown efficacy against clinically isolated strains of E. coli and pathogenic strains of S. aureus in animal models. Accordingly, chemically synthesized neoglycoconjugates based on PIA glycosides will be developed to serve as lead compounds for the development of new antibiotics as well as vaccines against biofilm dependent infections. Described in this thesis is a comprehensive study of the synthesis of PIA oligosaccharides and their deacetylated equivalents, the strategy for installing a stable linker on the free reducing oligosaccharide terminus and finally the conjugation to a model carrier protein for the development of potential neoglycoprotein vaccines.

ABSTRACT Objective: The objective of this review was to describe the effects of low carbohydrate diets in people with diabetes type 2. Methods: Review. An article search was made in the databases PubMed and Medline. The search resulted in 17 studies. After review and analysis of the studies main results five categories were highlighted; bloodsugar levels, HbA1c, weight, lipid levels and antidiabetic medications. Results: The results revealed that diets low in carbohydrates may reduce blood sugar levels, fasting blood glucose and HbA1c-levels in people with type 2 diabetes. A low carbohydrate diet may reduce body weight for people with type 2 diabetes, this was shown in 12 of 17 studies. Total cholesterol was reduced in six of 17 studies. A reduction of triglycerides was found in eight of 17 studies. LDL-cholesterol was reduced in four of the studies. HDL- cholesterol was increased in eight of 17 studies. Conclusion: Some of the studies showed positive effects of a low carbohydrate diet for people with type 2 diabetes, for instance improvements in blood sugar levels, bodyweight, total cholesterol, triglycerides, HDL-cholesterol and LDL-cholesterol. The study implies that a low carbohydrate diet may be an alternative for people with type 2 diabetes in short terms.Keywords: Diabetes type 2, diet, low-carbohydrate, ketogenic, HbA1c

The glycemic index (GI) of a pre-exercise meal has been shown to affect substrate oxidation during exercise and may influence exercise performance. Previous research in this area has focused on continuous, moderate intensity exercise. The purpose of this study was to examine the effects of low and high glycemic index (GI) pre-exercise meals on metabolism and performance during soccer-specific intermittent exercise. Thirteen trained male soccer players (22.3 ± 3.3 yrs) participated in four experimental trials in a repeated crossover design. Isocaloric low GI-high protein (lentils), high GI-high protein (potato + egg whites), or high GI-low protein (potato) meals were consumed two hours before a 90-minute treadmill soccer match simulation. A fasted control condition was also employed. Blood and expired gas samples were collected before and during exercise to assess markers of carbohydrate and fat metabolism. The distance covered on five 1-minute sprints (separated by 2.5 minutes of recovery) performed during the last 15 minutes of the match was used to assess performance. Serum insulin concentration at the start of exercise was higher in the high GI-low protein condition compared to all other conditions (p<0.001). During exercise, the rate of carbohydrate oxidation was significantly higher and the rate of fat oxidation was significantly lower in the high GI-low protein condition compared to control (p<0.05). The distance covered on sprints 1 and 2 was significantly greater in the low GI-high protein condition compared to control (p<0.05). The distance covered on sprint 2 was significantly greater in the high GI-high protein condition compared to control (p<0.05). There were no significant differences between conditions for sprints 3-5. Ratings of perceived exertion (RPE) throughout exercise were significantly lower in the low GI-high protein condition compared to both control and high GI-low protein conditions (p<0.05). It is concluded that low GI-high protein and high GI-high protein pre-exercise meals result in small improvements in initial repeated sprint performance compared to fasted control. Performance was not improved following a high GI-low protein pre-exercise meal compared to control, indicating that the increased carbohydrate oxidation and decreased fat oxidation in this condition may have negated any potential performance advantage of carbohydrate consumption. The lower RPE throughout exercise in low GI-high protein compared to control and high GI-low protein indicates a possible beneficial effect of consuming a low GI meal prior to soccer-specific intermittent exercise.

<p>Glucose metabolism remains an intensely explored topic of cancer biology since the initial discoveries of Otto Warburg nearly 80 years ago. Many solid tumors metabolize glucose primarily to lactate despite the availability of oxygen, revealing a dependence on glycolysis that may serve as a basis for targeted therapy. In particular, a diet devoid of carbohydrate may minimize the growth capabilities of glucose-dependent cancers. As our interests lie in prostate cancer, we examined whether a ketogenic diet devoid of carbohydrate (NCKD) would reduce the growth rate of tumors derived from human prostate cancer cell lines in a murine xenograft model.</p> <p>Our initial experiments utilized the LAPC-4 cell line, a human androgen-sensitive prostate cancer cell line, in a SCID-mouse xenograft model to determine the effects of an NCKD on tumor growth and animal survival relative to two other diets: (1.) a Western-type diet (WD) reflecting consumptions patterns of men diagnosed with prostate-cancer in the Western world and (2.) a low-fat diet (LFD) representing the present standard of care. Following this study, we conducted a second study utilizing a different human prostate cancer cell line (LNCaP) in order to assess whether our initial observations were robust across multiple prostate cancer tumor models and to also further explore the molecular underpinnings of our observations. Both studies revealed the NCKD leads to a reduction in tumor growth rate and greater overall mouse survival relative to the WD. In addition, the NCKD was equivalent in these parameters to the LFD. We also observed key associations between survival and extent of urinary ketosis as well as favorable changes in insulin and insulin-like growth factor-1 (IGF-1) and gene expression that would be predictive of prolonged survival in mice consuming the NCKD.</p> <p>We believe these data provide compelling evidence to consider a potential therapeutic role for dietary carbohydrate restriction in prostate cancer. We hope these results ultimately serve as a basis to conduct future clinical trials assessing whether dietary carbohydrate restriction, either alone or in combination with more conventional therapies, provides clinicians with an additional weapon against prostate cancer.</p> / Dissertation

Bird¡¦s nest is the saliva from the Colloclia that have a lot of mucin and sialic acid. However the sialic acid is an important intermediate of influenza virus infection. In 2006, Guo et al was confirmed the Inhibitory effect of bird¡¦s nest extract on influenza virus infection. But the anti-virus component has not been isolated and purified. First, the bird's nest used in this study was proved to be a pure bird¡¦s nest by SDS-PAGE, sialic acid content and protein content. Qualification analysis by SDS-PAGE showed sample possessed three bands at about 50 kDa, 100 kDa and 150 kDa, by Warren assay showed sample possessed a sialic acid content about 10% of dry weight, by Kjeldahl method showed sample possessed a protein content about 50% of dry weight, and by fluorescence analysis in 365 nm showed sample emitted a blue-white light. According to the criteria on previous studies, the sample was considered to be a real and pure bird¡¦s nest. Subsequently, the sample was applied to pronase digestion and then separated by Bio-gel P2 for size exclusion chromatography. All fractions were performed carbohydrate analysis including sialic acid. In lectin blotting, Maackia amurensis agglutinin (MAA) lectin can interact with the fraction that suggest the structure of sialic acid is sialic acid-2,3 galactose in bird¡¦s nest. And carbohydrate membrane array to demonstrate the influenza virus of H5N1 and H1N1 can bind the fractions that suggest the antivirus effect of bird¡¦s nest. These fractions will be further analyzed by neutralization assay to study their antivirus effects in vitro.