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Background: There are a number of biological pathways that affect our ingestive behaviours, including energy homeostasis, food reward, and taste. Given that carbohydrates such as sugars, provide energy and a sweet taste, examining candidate genes in each pathway may help explain differences in carbohydrate consumption behaviours. Objective: To determine whether variations in genes encoding a glucose transporter (GLUT2), a dopamine receptor (DRD2), and sweet taste receptor (TAS1R2) are associated with differences in sugar consumption in two distinct populations. Methods: Population 1 included diabetes-free young adults where dietary intake was assessed using a one month 196-item food frequency questionnaire (FFQ). Population 2 consisted of individuals with type 2 diabetes. Dietary intake was assessed using 3-day food records administered 2 weeks apart; food record 1 (FR1) and 2 (FR2). Subjects were genotyped for the Thr110Ile variation in GLUT2 (n1=587; n2=100), the C957T variation in DRD2 (n1=313; n2=100), and the Ser9Cys and Ile191Val variations in TAS1R2 (n1=1037; n2=100) using real-time PCR. Results: In comparison to individuals homozygous for the GLUT2 Thr allele, consumption of sugars was higher among Ile carriers in population 1 (133 ± 5 vs 118 ± 3 g/d, p=0.006) and population 2 on two separate food records (FR1: 112 ± 9 vs 87 ± 5 g/d, p=0.02; FR2: 105 ± 8 vs 78 ± 4 g/d, p=0.002). For the C957T variation in population 1, we detected a significant DRD2xSex interaction with the consumption of sucrose decreasing with each T allele among men (p=0.03) and a heterosis mode of inheritance among women where heterozygotes consumed the most (p=0.01). For TAS1R2, we detected a significant TAS1R2xBMI interaction and among overweight individuals, carriers of the Val allele consumed less sugars than those with the Ile/Ile genotype (103 ± 6 vs122 ± 6 g/d, p=0.01). In population 2, carriers of the Val allele consumed less sugars than individuals with the Ile/Ile genotype (83 ± 6 vs 99 ± 6 g/d, p=0.04) on FR2. Conclusions: Our findings demonstrate that genetic variation in GLUT2, DRD2 and TAS1R2 affect habitual sugar consumption and suggest that selection of dietary sugars can be influenced by different biological pathways.

Artemisinin (AN), a sesquiterpene, derived from the herb, Artemisia annua is the most widely used anti-malarial compound. Current production is insufficient to meet the growing demand for this important drug. Many experiments have been done to try and deduce what factors may be important to increased yield. Here is is shown that many disparate phenomena known to induce AN production may be linked under the umbrella of reactive oxygen species (ROS). To that end, the metabolite and transcriptional changes associated with the transition from vegetative growth to flowering have been investigated. In addition, the role that exogenous sugars play in modulating these same factors has been explored in young seedlings. Lastly, exposure to DMSO was shown to increase AN production and that it may be linked to ROS. These combined results wered further explored to determine the effects of direct ROS elicitation and subsequent quenching on the production of AN and related metabolites. Information gained here supported a new alternative hypothesis for the role of ROS in AN production, one in which hydrogen peroxide may be controlling the balance of deoxyartemisinin (deoxyAN) and AN.

Severe dehydration is lethal for most biological species, however there are a number of organisms which have evolved mechanisms to avoid damage during dehydration. One of these mechanisms is the accumulation of small solutes (e.g. sugars), which have been shown to preserve membranes by inhibiting deleterious phase changes at low hydration. Specifically, sugars reduce the gel to fluid phase transition temperatures of model lipid/water mixtures. However, there is debate about the precise mechanism, the resolution of which hinges on the location of the sugars. An experimental investigation into the effects of small solutes on the phase behaviour of phospholipid membranes is presented in order help identify the mechanisms by which solutes facilitate desiccation tolerance. Differential Scanning Calorimetry (DSC) was used to determine the first comprehensive phase diagram for the synthetic phospholipid DPPC over a wide range of hydration and solute molar ratios between 0.1 and 1.0 sugars per lipid. Over the same range of hydrations and solute concentrations Small Angle X-Ray Scattering (SAXS) was used to measure the structural parameters of the membrane bilayers necessary to determine both the phase of the membrane lipids and the location of the solutes. SAXS was also used to conduct the first comprehensive study of the effect of solutes on the kinetics of the fluid - gel transition of DPPC over a range of both hydration and solute concentration. Finally, contrast variation Small Angle Neutron Scattering (SANS) was used to quantitatively determine the location of the solutes. Data from these complimentary techniques are presented which show a monotonic relationship between both transition temperature and repeat spacing with respect to solute concentration. This relationship exists between solute:lipid molar ratios between 0.1 to approximately 0.5, after which higher concentrations of solute are shown to have no further effect on either the bilayer repeat spacing or transition temperature. It is proposed that the exclusion of small solutes into sugar/water micro-phases external to the bilayer can account for this behaviour. A theoretical model previously used to describe membrane phase behaviour at low hydrations is modified to account for the presence of solutes between membrane bilayers. This model is shown to be in quantitative agreement with the experimental data up until approximately 0.5 sucrose molecules per lipid, the point of solute exclusion. Once exclusion is taken into account, the model is quantitative over the whole range of sugar concentrations.

Carbohydrates are known to play a large number of significant roles in various biological and pathological processes such as cancer metastasis and cellular communication. This is because of their ability to bind a wide range of hosts within the human body such as proteins and viruses. Due to these important interactions, carbohydrate sensing has long been a main focus of research. These research strategies have included the use of aptamers, non-covalent interactions, and boronic acid-based receptors. Boronic acid-based sensors are of particular interest due to their selectivity for 1,2- or 1,3-diols. Within these boronic acid-based studies, a large variety of techniques were employed for detection including different fluorescent, electrochemical, polymeric, and colorimetric studies, as well as various surface bound sensors. One type of technique that has rarely been applied is Surface Enhanced Raman Spectroscopy or SERS. This strategy would be beneficial as it provides information about functional groups, which would aid in the identification of the bound sugar. In this thesis, we present work based on the development of a boronic acid-based carbohydrate receptor that will be used to study carbohydrate binding through SERS. The receptor design includes an aryl boronic acid for carbohydrate recognition, a nitrogen atom in close proximity to the boron center to enhance binding, and a terminal thiol for attachment to a metal surface for SERS. This sensor will be used to study the binding of different saccharides for sensing applications.

Urban forests grow in stressful environments that can have negativerepercussions on tree energy reserves. The goal of this research was to evaluate theimpact of exogenously applied carbohydrates on growth and vitality of live oaks(Quercus virginiana P. Miller). An initial study focused on carbohydrate partitioningrevealed that annual mean glucose concentration in leaf tissues (49.55 mg·g-1 DW) wasalmost double that in twigs, trunks, or roots. Starch concentrations in roots and trunks(38.98 and 38.22 mg·g-1 DW of glucose, respectively) were higher during the dormantseason and approximately three times the concentrations found in other tissues. Aninvestigation of the effects of exogenous soil applications of glucose and starch on soilmicrobial activity revealed no significant differences using recoverable viable microbes.However, soil respiration was significantly increased (P<0.05) by glucose a week afterapplication, while higher starch concentrations (120 g·L-1) significantly increased(P<0.05) soil respiration after the fourth week. Although tree soil drenched withcarbohydrates in a different study showed significantly (P<0.05) greener leaf color, higher chlorophyll fluorescence, and increased soil respiration at higher concentrationsof starch (120 g·L-1), no significant differences were observed in photosynthesis or trunk,canopy, or root growth. Analysis of 13C signatures was unable to detect uptake ofexogenous carbohydrates. For trunk-injected trees with glucose and sucrose, trunkgrowth was significantly (P<0.05) increased by carbohydrate supplementation.Differences were also found in twig glucose content, root starch content, and chlorophyllfluorescence among overall concentration means. A study to compare field diagnostictools with carbohydrate laboratory analysis established that a portable blood glucosemeter can be used to measure glucose content in trees. However, ohmmeter,refractometer, chlorophyll fluorescence spectrometer, and iodine staining results did notcorrelate well with laboratory analysis of carbohydrate concentrations. Results fromthese studies reveal that soil applied carbohydrates can greatly increase soil microbialactivity, provide evidence that trunk-injected carbohydrates may improve growth andvitality of live oaks, and provide a new field diagnostic tool to increase the efficiency ofmeasuring carbohydrates in trees.

Cyclopropanes are strained molecules and undergo reactions, for example, through ring opening and rearrangements. Preparative methods and reactivities of cyclopropanes are known widely in organic synthesis. The high reactivities inherent in cyclopropanes allow them to be valuable building blocks in organic synthesis. The combination of cyclopropanes and carbohydrates has been explored in recent years. Carbohydrates, the naturally-occurring members of chiral pool, are attractive platforms for asymmetric synthesis. Cyclopropanation of, for example, unsaturated sugars affords [4.1.0] bicyclic systems, thereby combining the high reactivities of cyclopropanes together with optical purities of sugars. Chapter 1 of the Thesis describes (i) various types of cyclopropane ring opening reactions in general and (ii) known reactions of cyclopropanes in carbohydrates relevant to the work presented in the Thesis. Seven-membered cyclic sugars, namely, septanoses and septanosides, are less commonly known sugar homologues. Synthesis of septanoses arise interest, due to their configurational and conformational features and the attendant possibilities to explore their chemical, physical and biological properties. In a programme, it was desired to identify a new methodology for synthesis of septanosides. It was envisaged that 2-hydroxy glycals, namely, oxyglycals, would form as suitable substrates for ring expansion, leading to the formation of septanoside derivatives that are retained with hydroxyl groups in each carbon of the septanoside. In the event, a new methodology was identified. A carbene insertion of an oxyglycal substrate, nucleophilic ring opening of the cyclopropyl moiety, oxidation and reduction reactions were identified to expand the six membered pyranoses to seven membered septanosides (Scheme 1). The methodology was established through preparation of two configurationally different septanosides, namely, the methyl α-D-glycero-D-talo-septanoside and methyl α-D-glycero-L-altro-septanoside from D-glucose and D-galactose, respectively. Chapter 2 presents details of the methodology and the preparation of septanosides from precursors oxyglucal and oxygalactal. Scheme 1 Continuing the efforts to extend the methodology, preparation of a variety of septanosides, using phenoxides, sugars and azide were undertaken. It was found that ring opening with sugars were highly stereoselective, leading to an exclusive formation of the -anomer of sugar oxepines, whereas, the phenoxides and azide led to a mixture of anomers of the corresponding oxepines, in a ~1:1 ratio (Scheme 2). Scheme 2 An important observation was -anomer of the oxepine derived intermediates, having diketo-functionalities, underwent NaBH4 mediated conversion to diols with higher diastereoselectivities at the newly generated stereo-centers, whereas the -anomers lacked to retain the diastereoselectivities, in the case of aryl septanosides. This part of work consolidated further the generality of the oxyglycal ring-expansion method to prepare septanosides, possessing different substituents at their reducing ends. Chapter 3 describes the details of syntheses and characterization of various aryl septanosides, septanoside disaccharides and azido-septanoside derivatives. It was planned further to synthesize septanoside containing di-and trisaccharides from naturally-occurring disaccharides, through the oxyglycal route. Oxyglycals, derived from lactose and maltose, were expanded to septanoside-containing trisaccharides through a ring expansion method. Thus septanosides incorporated disaccharides and trisaccharides, with 6-7, 6-7-5 and 6-7-6 ring sizes, were prepared through the ring expansion method. The reaction not only led to a ring expansion, but also, to a concomitant glycoside formation, in a stereoselective manner (Scheme 3). Scheme 3 A conformational analysis of the galacto-septano-glucopyrano-configured 6-7-6 trisaccharide was undertaken with aid of NMR spectroscopy and computational methods. Spatial distances from NMR experiments were utilized while performing molecular dynamics with AMBER* force field and further optimizations using B3LYP/6-31+G* level. The study showed that septanoside ring in the trisaccharide adopted twist-chair conformation O,1TC5,6, as shown in Figure 1. Chapter 4 describes synthesis of septanoside containing di-and trisaccharides and conformational analysis of a 6-7-6 trisaccharide, through solution phase and computational methods. An effort was pursued to prepare septanoside-based amphiphiles with varying alkyl chain lengths, using our newly established methodology and to study their amphiphilicities. A series of septanoside amphiphiles, having C10 to C18 alkyl groups, were prepared as their -anomers as shown in Figure 2. The amphiphilic behavior of the alkyl septanosides was assessed through studies of their liquid crystalline (LC) properties. The LC properties were evaluated using polarizing optical microscopy, differential scanning calorimetry and powder X-ray diffraction methods. All the septanoside amphiphiles exhibited a smectic A phase in general. DSC thermograms showed crystal-crystal and crystal-mesophase phase transitions. Powder X-ray diffraction studies allowed to identify the lamellar structuring of the smectic A phase. Further, two distinct two layer spacings were observed. Such an observation is un-usual in carbohydrate liquid crystals. Chapter 5 details of synthesis and studies of the mesomorphic behavior of septanoside amphiphiles. In summary, the Thesis establishes a new route to synthesize septanoside derivatives, from oxyglycal sugar derivatives. Ring expansion of a pyranoside to a septanoside was achieved through key reactions of a cyclopropanation, ring opening, oxidation and reduction. Methyl α-D-glycero-septanoside derivatives were synthesized, from the corresponding oxyglycals. Cyclopropane ring opening ability of various nucleophiles were studied, it was found that ring-opening reactions with phenols, sugars, and azides are effective, which facilitated the synthesis of various aryl, glycosyl and azido-substituted septanosides. Synthesis of septanosides incorporated with di-and trisaccharides were accomplished. The detailed conformational analysis studies showed that the septanoside adopted twist-chair conformation in a trisaccharide molecule. Preparation and studies of septanoside based amphiphiles and their mesophase behavior were also accomplished. Overall, the studies presented in the Thesis provide a new insight to ring expanded sugars. The salient features of the present method are that the intermediates such as the seven membered vinyl halides, vinyl ethers, the diketones and the diols are potential sites for many other functionalizations. These features can be explored further in functionalizing the newly formed septanosides. (For structural formula pl see the pdf file)

Urban forests grow in stressful environments that can have negativerepercussions on tree energy reserves. The goal of this research was to evaluate theimpact of exogenously applied carbohydrates on growth and vitality of live oaks(Quercus virginiana P. Miller). An initial study focused on carbohydrate partitioningrevealed that annual mean glucose concentration in leaf tissues (49.55 mg·g-1 DW) wasalmost double that in twigs, trunks, or roots. Starch concentrations in roots and trunks(38.98 and 38.22 mg·g-1 DW of glucose, respectively) were higher during the dormantseason and approximately three times the concentrations found in other tissues. Aninvestigation of the effects of exogenous soil applications of glucose and starch on soilmicrobial activity revealed no significant differences using recoverable viable microbes.However, soil respiration was significantly increased (P<0.05) by glucose a week afterapplication, while higher starch concentrations (120 g·L-1) significantly increased(P<0.05) soil respiration after the fourth week. Although tree soil drenched withcarbohydrates in a different study showed significantly (P<0.05) greener leaf color, higher chlorophyll fluorescence, and increased soil respiration at higher concentrationsof starch (120 g·L-1), no significant differences were observed in photosynthesis or trunk,canopy, or root growth. Analysis of 13C signatures was unable to detect uptake ofexogenous carbohydrates. For trunk-injected trees with glucose and sucrose, trunkgrowth was significantly (P<0.05) increased by carbohydrate supplementation.Differences were also found in twig glucose content, root starch content, and chlorophyllfluorescence among overall concentration means. A study to compare field diagnostictools with carbohydrate laboratory analysis established that a portable blood glucosemeter can be used to measure glucose content in trees. However, ohmmeter,refractometer, chlorophyll fluorescence spectrometer, and iodine staining results did notcorrelate well with laboratory analysis of carbohydrate concentrations. Results fromthese studies reveal that soil applied carbohydrates can greatly increase soil microbialactivity, provide evidence that trunk-injected carbohydrates may improve growth andvitality of live oaks, and provide a new field diagnostic tool to increase the efficiency ofmeasuring carbohydrates in trees.