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Taste aversion to sugars by the gerbil.

Some conditioned taste aversion experiments were undertaken to determine how the gerbil responds to disaccharides, monosaccharides and polyols. We observed the following: animals taught an aversion to 0.1 M sucrose generalized the avoidance to most sugars, the exception being galactitol; animals taught to avoid 0.01 M hydrochloric acid generalized the avoidance towards lactose, cellobiose, maltitol, methyl alpha-D-galactopyranoside, methyl alpha-D-mannopyranoside, methyl beta-D-glucopyranoside, and glycerol; animals taught to avoid 0.001 M quinine . HCl generalized the avoidance towards methyl alpha-D-glucopyranoside, methyl-beta-D-glucopyranoside, glycerol, ethylene glycol and erythritol. In no case did animals taught to avoid 0.1 M sodium chloride avoid any of the sugars. Moreover, it was observed that the gerbil's behavior with most reducing sugars was different than with equivalent methyl glycosides. For example, animals that were taught to avoid sucrose generalized the avoidance towards reducing sugars, such as, D-galactose, D-glucose, and D-mannose. However, the methyl glycosides, such as methyl alpha-D-glucopyranoside, methyl beta-D-glucopyranoside, methyl alpha-D-galactopyranoside and methyl alpha-D-mannopyranoside, in addition to being avoided by animals taught to avoid sucrose, were also avoided by animals taught to avoid quinine . HCl or hydrochloric acid. In addition, we have observed that the control animals consumed differing amounts of sugars and have concluded, therefore, that the sugars were not equally pleasant despite our attempt to use concentrations which produced equally intense neural responses in the gerbil's chorda tympani nerve.

Animals↗

The use of sugar-coated beads to study phagocytosis in normal and dystrophic retina.

In this study, a technique is described for coupling the sugars to carboxylated latex beads for use in phagocytic studies of dystrophic and normal retinal pigment epithelium. The presence and distribution of the sugars, mannose and fucose, on the bead surfaces are visualized by transmission electron microscopy using the lectin-Ferritin conjugates. Concanavalin A-Ferritin (Con A-Fe) and Ulex-europeus-Ferritin (Ulex-Fe), respectively. Both lectin-Ferritin conjugates label the surfaces of the sugar coated beads. Control experiments to determine the extent of nonspecific lectin-Ferritin binding using Con A-Fe with fucose-coated beads, Ulex-Fe with mannose-coated beads, or beads without coupling reagent show little to no labeling. Lectin-Ferritin conjugates are also incubated with the appropriate sugar-coated beads and their competing sugars and little to no labeling is observed. Our data shows that when sugar-coated beads are presented to retinal pigment epithelial explants, differences in phagocytic uptake are observed. Uncoated beads are avidly taken up by normal and dystrophic tissue, but fucose-coated beads are not taken up by either. Twice as many mannose beads are phagocytized by the normal as compared to dystrophic tissue. It remains to be established whether these differences in bead uptake indicate a difference in ligand-receptor interactions between normal and dystrophic retinal pigment epithelium.

Animals↗

Glycosylation of the variable region of immunoglobulin G--site specific maturation of the sugar chains.

The structure of the N-linked sugar chains attached to three IgG antibodies, identical in amino acid sequence except for the changes required to introduce the carbohydrate addition sites, has been determined. All three antibodies are specific for dextran but differ in their ability to bind antigen. The heavy chains with a murine variable region (V region) attached to the human gamma 4 constant region were expressed in a murine hybridoma synthesizing the specific light chain. In addition to the glycosylation site in the Fc portion, each antibody has a different glycosylation site in the second complementarity determining region (CDR2) of the heavy chain (Asn54, Asn58, or Asn60). The sugar chains were released from purified Fab and Fc fragments by hydrazinolysis and converted to radioactive oligosaccharides by reduction with sodium borotritide. The structures of these radioactive oligosaccharides were determined by a combination of sequential exoglycosidase digestion and Bio-Gel P-4 and lectin column chromatography. For all three antibodies, the carbohydrate attached to the Fc portion was a mixture of complex-type biantennary sugar chains. The variable region carbohydrate structures attached at Asn54 and Asn58 were also complex-type but more highly sialylated than were the Fc-associated sugars. Moreover, unlike the Fc-associated sugars, a significant population of Fab-associated sugars contained a Gal alpha 1-->3 residue as a non-reducing terminus. In contrast, the carbohydrate attached at Asn60 was a high mannose structure. These results demonstrate that slight changes in the position of carbohydrate attachment within CDR2 of the variable region of the heavy chain can substantially alter carbohydrate processing and that complex-type carbohydrates contained within the same polypeptide chain can have different structures.

Amino Acid Sequence↗

Inhibition of the serosal sugar carrier in isolated intestinal epithelial cells by saccharin.

Isolated intestinal cells accumulate certain monosaccharides via an Na+-dependent, active transport system localized in the brush-border membrane, and release sugar molecules at the basolateral boundary via a facilitated diffusion, passive system. Work described here indicates that sodium saccharin (25-130 mM) has little if any direct effect on the active transport system, but that the passive transport system is inhibited by saccharin. A short period of exposure (10-60 min) is required for expression of the effect, which is detectable at saccharin concentrations as low as 10 mM. At 100 mM-sodium saccharin, as much as 50% inhibition occurs. Saccharin also appears to act as a weak metabolic inhibitor. The basis of the 'non-specific' effect is not understood, but it can compromise the capacity of the epithelial cells to form sugar gradients. When a sugar is accumulated that satisfies both transport systems (for example 3-O-methylglucose) the effect of saccharin on the passive transport system is the predominant one, and the cells establish a higher sugar gradient than that observed in the absence of saccharin. The 'non-specific' metabolic effect is manifested as an inhibition of sugar gradient formation when sugars that satisfy only the active system (such as alpha-methylglucoside) are accumulated.

Animals↗

Bioavailability of oxalic acid from spinach, sugar beet fibre and a solution of sodium oxalate consumed by female volunteers.

Oxalate bioavailability from sugar beet fibre (40 g), spinach (25 g) and a solution of sodium oxalate (182 mg) was tested in nine women using a triplicated 3 x 3 Latin square arrangement. Each test substance provided 120 mg oxalic acid. Throughout the study the volunteers consumed a control diet and the test substances were administered at breakfast on specified days. After an initial 2-day control period, oxalate was administered in three test periods that consisted of one test day followed by one control day. Urine collected during 24-hr periods was analysed daily for oxalate. Oxalate excretion did not differ among the five control days and was not increased significantly following the ingestion of sugar beet fibre by the volunteers. Oxalate excretion was greater (P less than 0.0001) for the mean of the spinach and sodium oxalate solution diets than for the mean of the sugar beet fibre and control diets. Oxalate bioavailability from sugar beet fibre was 0.7% compared with bioavailabilities of 4.5 and 6.2% for spinach and oxalate solutions, respectively. The low bioavailability of oxalate from sugar beet fibre may be attributable to its high ratio of minerals (calcium and magnesium) to oxalate, its complex fibre matrix or the loss of the soluble oxalate during processing of sugar beets.

Administration, Oral↗

Sugar interaction with biologically active cis-PtCl2(NH3)2 (anticancer) and its inactive trans isomer.

The interaction of D-glucuronic and D-gluconic acids with cis- and trans-PtCl2(NH3)2 (cisplatin and transplatin) has been investigated in aqueous solution and solid complexes of the type cis-[PtL(NH3)2]L.H2O and trans-[PtL2(NH3)2]L.H2O, where L = D-glucuronate or D-gluconate anions, are isolated and characterized by means of Fourier transform-infrared and 1H-NMR spectroscopy, and molar conductivity and X-ray powder diffraction measurements. Spectroscopic and other evidence indicated that the sugar anions bind monodentately in trans-[PtL2(NH3)2].H2O and bidentately in cis-[PtL(NH3)2]L.H2O complexes through the carboxylate oxygen atoms and other sugar donor groups. The strong sugar intermolecular hydrogen-bonding network is altered to that of the sugar-OH...NH3(H2O)...OH-sugar, upon platinum-ammine interaction. The D-glucuronate anion has the beta-anomer configuration both in the free salt and in these platinum-sugar complexes.

Cisplatin↗

Role of calcium influx in the regulation of sugar transport in resting left atrial muscle.

The role of external Ca2+ in the regulation of sugar transport in isolated resting atria of rats and guinea pigs was studied by measuring the tissue/medium distribution of 14C-labelled 3-methylglucose and of 45Ca. Omission of Ca2+ from the medium strongly antagonized the stimulation of sugar transport by insulin, hyperosmolarity (100 mM mannitol) or Na+-pump inhibition (K+-free medium). Basal sugar transport was not affected by Ca2+ omission but was increased when 0.5 mM EGTA was also added. The Ca2+ antagonist drug D-600 decreased 45Ca influx and also inhibited sugar-transport stimulation by the above 3 treatments in the rat, while in the guinea pig it antagonized only insulin-stimulated transport. The stimulation of sugar transport by a high adrenaline concentration and the inhibition by a low concentration were both antagonized by Ca2+ omission or D-600. The results illustrate the important role of Ca2+ influx in the control of sugar transport by hormonal and other modulators and are consistent with the hypothesis that cytoplasmic Ca2+ regulates glucose transport in muscle.

Animals↗

Distribution of O-linked sugar units on hCG and its free alpha-subunit.

hCG, a glycoprotein hormone produced by the trophoblast in pregnancy, is composed of two dissimilar subunits, alpha and beta, joined non-covalently. hCG has four O-linked sugar units, all attached to the beta-subunit. The trophoblast also produces a free form of the alpha-subunit, which unlike the alpha-component of hCG, can contain an O-linked sugar unit. The structures of the O-linked sugar units were examined. Four structures were identified on urinary hCG. A hexasaccharide, NeuAc alpha 2-3Gal beta 1-3(NeuAc alpha 2-3Gal beta 1-4GlcNAc beta 1-6)GalNAc- accounting for 13%, a tetrasaccharide, NeuAc alpha 2-3Gal beta 1-3(NeuAc alpha 2-6)GalNAc-, for 34%, a trisaccharide, NeuAc alpha 2-3Gal beta 1-3GalNAc-, for 43% and a disaccharide, NeuAc alpha 2-6GalNAc- for 10% of the total O-linked sugar structures. Similar mixtures were found on peptides containing one, three or four sugar units suggesting a random distribution among attachment sites. The distribution of O-linked sugar structures on hCG and free alpha-subunit from trophoblast explant cultures was compared. The mixture of structures attached at the single site on the free alpha-subunit paralleled that at the four sites on the hCG.

Carbohydrate Conformation↗

Starch and sugar tastes in rodents: an update.

Rats are strongly attracted to the sweet taste of sugar. Recent behavioral studies demonstrate that rats also have a well-developed taste for starch-derived polysaccharides (e.g., Polycose). In fact, rats prefer Polycose to sucrose and other sugars at low concentrations. Polycose appetite develops at a very young age (9 days) and, thus, appears to be innate. The results of conditioned taste aversion tests suggest that rats taste Polycose as qualitatively different from sucrose. Recent electrophysiological findings support the idea that rodents have separate taste channels for polysaccharides and sugars. In particular, copper chloride suppresses the chorda tympani nerve response to sucrose and other sugars but has minimal effect on the neural response to Polycose. Also, Polycose evokes a profile of neural activity in the nucleus tractus solitarius that differs substantially from that produced by sucrose. Preliminary results indicate that polysaccharide and sugar tastes also differ in their metabolic consequences, i.e., unlike sugars, Polycose does not elicit a cephalic phase insulin response. The presumed function of polysaccharide taste is to facilitate the identification of starch-rich foods. Recent findings demonstrate that rats can readily detect starch even at low concentrations, but whether polysaccharide taste receptors or other orosensory receptors mediate this response remains to be clarified.

Animals↗

A hexokinase with broad sugar specificity from a thermophilic bacterium.

A recombinant thermophilic Thermus caldophilus GK24 hexokinase, one of the ROK-type (repressor protein, open reading frames, and sugar kinase) proteins, exists uniquely as a 120 kDa molecule with four subunits (31 kDa), in contrast to eukaryotic and bacterial sugar kinases which are monomers or dimers. The optimal temperature and pH for the enzyme reaction are 70-80 degrees C and 7.5, respectively. This enzyme shows broad specificity toward glucose, mannose, glucosamine, allose, 2-deoxyglucose, and fructose. To understand the sugar specificity at a structural level, the enzyme-ATP/Mg2+-sugar binding complex models have been constructed. It has been shown that the sugar specificity is probably dependent on the interaction energy occurred by the positional proximity of sugars bound in the active site of the enzyme, which exhibits a tolerance to modification at C2 or C3 of glucose.

Adenosine Triphosphate↗

Induction of BiP by sugar independent of a cis-element for the unfolded protein response in Arabidopsis thaliana.

BiP is a molecular chaperone induced in the unfolded protein response (UPR). In mammalian cells, BiP is induced by glucose starvation when it is called glucose-regulated protein 78 (GRP78). In Arabidopsis thaliana, however, we demonstrated that BiP transcripts decreased with sugar depletion and increased with sugar addition. Transcripts for beta-glucuronidase (GUS) driven by BiP promoter respond to tunicamycin and sugar, being similar with endogenous BiP transcripts in transgenic A. thaliana. When GUS was regulated by P-UPRE, a cis-element responsible for the UPR identified in BiP promoter, GUS transcripts were accumulated by sugar starvation. Subsequently, transgenic A. thaliana harboring luciferase (LUC) gene regulated by P-UPRE was analyzed. Sugar depletion also increased LUC activity. It is concluded that BiP is induced by sugar independent of the cis-element responsible for the UPR.

Arabidopsis↗

Conventional transport assays underestimate sugar transport rates in human red cells.

The time course of protein-mediated 3-O-methylglucose uptake by human red cells and by red cell ghosts containing or lacking 4 mM MgATP was measured at ice temperature and sub-saturating sugar levels by conventional sampling procedures and at 20 degrees C by use of a quench-flow apparatus. The temporal resolution of the quench-flow apparatus (as fast as 5-ms sample times) was confirmed by analysis of alkaline hydrolysis of dinitrophenolacetate. Red cell sugar uptake at 4 degrees C is consistent with two processes [fast (tau = 120 s) and slow (tau = 1100 s)] that occur in series. Intracellular ATP increases the size and the rate of equilibration of the fast compartment and slows the rate of filling of the slow compartment. Red cell ghost volume and protein content are unaffected by lysis/resealing in the presence of ATP. Uptake at 20 degrees C is also consistent with two processes [fast (tau = 10 ms) and slow (tau = 15 s)] that occur in series. ATP increases the size of both compartments and the rate of filling of the small compartment at 20 degrees C. Preliminary estimates indicate that the sugar uptake capacity of human red cells at 20 degrees C is underestimated by as much as 8-fold by measuring sugar uptake over 2 s vs. 26 ms. We discuss the implications of multiphasic sugar uptake in the context of models for protein-mediated sugar transport.

3-O-Methylglucose↗

Dilute acid hydrolysis of presscakes from silage and grass to recover hemicellulose-derived sugars.

Grass and silage presscakes from various types of raw materials were hydrolyzed with dilute acid at moderate conditions to recover hemicellulose-derived sugars. Extracting the material with cold water prior to hydrolysis significantly increased the yield. The poor performance without extraction was due to the high buffer capacity of the material. The best results were obtained with extracted grass and silage from a permanent pasture and extracted clover/grass silage. The highest observed sugar yield was 16.43 g/100 g dry-matter, which represents approximately 25% of the available sugars and 60% of the hemicellulose fraction. Including the soluble sugar oligomers, the yields were even higher (up to 20 g/100 g dry-matter). A statistical experiment design with extracted clover/grass silage was performed to estimate the effects of temperature, time, and dry-matter concentration. Acid and dry-matter concentration had the highest effect on sugar yield, whereas temperature and acid concentration were mainly responsible for forming sugar degradation products. These findings agree with recent kinetic theories. Yields in this experiment were comparable to those of other lignocellulosic materials.

Carbohydrates↗

Refining sweet sorghum to ethanol and sugar: economic trade-offs in the context of North China.

Reducing the use of non-renewable fossil energy reserves together with improving the environment are two important reasons that drive interest in the use of bioethanol as an automotive fuel. Conversion of sugar and starch to ethanol has been proven at an industrial scale in Brazil and the United States, respectively, and this alcohol has been able to compete with conventional gasoline due to various incentives. In this paper, we examined making ethanol from the sugar extracted from the juice of sweet sorghum and/or from the hemicellulose and cellulose in the residual sorghum bagasse versus selling the sugar from the juice or burning the bagasse to make electricity in four scenarios in the context of North China. In general terms, the production of ethanol from the hemicellulose and cellulose in bagasse was more favorable than burning it to make power, but the relative merits of making ethanol or sugar from the juice was very sensitive to the price of sugar in China. This result was confirmed by both process economics and analysis of opportunity costs. Thus, a flexible plant capable of making both sugar and fuel-ethanol from the juice is recommended. Overall, ethanol production from sorghum bagasse appears very favorable, but other agricultural residues such as corn stover and rice hulls would likely provide a more attractive feedstock for making ethanol in the medium and long term due to their extensive availability in North China and their independence from other markets. Furthermore, the process for residue conversion was based on particular design assumptions, and other technologies could enhance competitiveness while considerations such as perceived risk could impede applications.

Bioreactors↗

Transformations of bioactive peptides in the presence of sugars--characterization and stability studies of the adducts generated via the Maillard reaction.

Glycation of biomolecules, such as proteins, peptide hormones, nucleic acids, and lipids, may be a major contributor to the pathological manifestations of aging and diabetes mellitus. These nonenzymatic reactions, also termed the Maillard reaction, alter the biological and chemical properties of biomolecules. In order to investigate the effect of various reducing sugars on the products formed from small bioactive peptides (Tyr-Gly-Gly-Phe-Leu, Tyr-Gly-Gly-Phe-Leu-NH2, Tyr-Gly-Gly-Phe-Leu-OMe, Tyr-Gly-Gly-Phe, and Tyr-Gly-Gly), model systems were prepared with glucose, mannose or galactose. Peptide-sugar mixtures were incubated at 37 or 50 degrees C in phosphate-buffered saline, pH 7.4, or in methanol. The extent of glycation was determined periodically by RP HPLC. All sugar-peptide mixtures generated two different types of glycation products: N-(1-deoxy-ketos-1-yl)-peptide (Amadori compound) and the imidazolidinone compound substituted by sugar pentitol and peptide residue. The amount and distribution of peptide glycation products depended on the structure of the reactants, and increased in both concentration- and time-dependent manner in relation to exposure to sugar. Additionally, the rate of hydrolysis of glucose-derived imidazolidinone compounds, obtained either from leucine-enkephalin (1) or its shorter N-terminal fragments 2 and 3, was determined by incubation at 37 degrees C in human serum. These results revealed that imidazolidinones obtained from glucose and small peptides are almost completely protected from the action of enzymes in serum, the predominant route of degradation being spontaneous hydrolysis to initial sugar and peptide compound.

Carbohydrate Metabolism↗

Physiological differences between two sugar-sensitive neurons in the galea and the maxillary palp of the spruce budworm larva Choristoneura fumiferana (Clem.) (Lepidoptera: Tortricidae).

The L1 contact-chemoreceptor sensillum on the maxillary palp of the spruce budworm larva Choristoneura fumiferana (Clem.) (Lepidoptera: Tortricidae) was examined electrophysiologically for its responses to stimulation by various pyranose and furanose sugars. The results were compared to those from previous work on the sugar-sensitive neuron of the LST sensillum on the galea. We show that the L1 contact-chemoreceptor sensillum contains one sugar-sensitive neuron with furanose but no pyranose sites. It has response characteristics that differ from those of the sugar-sensitive neuron in the LST. Behavioural 2-choice feeding experiments show that, even with both known sugar-sensitive neurons disabled, larvae can still discriminate between disks treated with either distilled water or alpha-D-glucose. We conclude that the epipharyngeal sensilla must thus also contain a sugar-sensitive neuron.

Animals↗

An unusual sugar conformation in the structure of an RNA/DNA decamer of the polypurine tract may affect recognition by RNase H.

Retroviral conversion of single-stranded RNA into double-stranded DNA requires priming for each strand. While host cellular t-RNA serves as primer for the first strand, the viral polypurine tract (PPT) is primer for the second. Therefore, polypurine tracts of retroviruses are essential for viral replication by reverse transcriptase (RT). These purine tracts are resistant to cleavage during first strand synthesis. In obtaining the primer for second strand synthesis, the RNase H function of RT must cleave the PPT exactly for in vivo transcription to proceed efficiently and proper integration to occur. At the RNase H active site the protein makes contacts primarily along the backbone, with hydrogen bonds to the sugar-phosphate oxygen atoms. A high-resolution structure (1.10A) of the first ten base-pairs of the RNA/DNA hybrid PPT, r-(c-a-a-a-g-a-a-a-a-g)/d-(C-T-T-T-T-C-T-T-T-G), contains the highly deformable r-(a-g-a) steps found in retroviral polypurine tracts. This r-(a-g-a) motif is utilized in the "unzipping" or unpairing of bases that occurs when RT binds a malleable PPT. Another unusual feature found in our high-resolution PPT structure is the sugar switch at RNA adenine 2. All the RNA sugars are the expected C3'-endo, except sugar 2, which is C2'-endo, characteristic of B-form sugars. This local A-to-B conversion adversely affects the pattern of hydrogen bonds from protein to sugar-phosphate backbone, disrupting the catalytic site. Disruption could cause the enzyme to pause at the 5'-end of the PPT, leaving it intact. Pyrimidine-purine (YR) steps are most deformable and the T-A step especially can undergo A-to-B transitions readily.

Base Sequence↗

Protein-DNA hydrophobic recognition in the minor groove is facilitated by sugar switching.

Information readout in the DNA minor groove is accompanied by substantial DNA deformations, such as sugar switching between the two conformational domains, B-like C2'-endo and A-like C3'-endo. The effect of sugar puckering on the sequence-dependent protein-DNA interactions has not been studied systematically, however. Here, we analyzed the structural role of A-like nucleotides in 156 protein-DNA complexes solved by X-ray crystallography and NMR. To this end, a new algorithm was developed to distinguish interactions in the minor groove from those in the major groove, and to calculate the solvent-accessible surface areas in each groove separately. Based on this approach, we found a striking difference between the sets of amino acids interacting with B-like and A-like nucleotides in the minor groove. Polar amino acids mostly interact with B-nucleotides, while hydrophobic amino acids interact extensively with A-nucleotides (a hydrophobicity-structure correlation). This tendency is consistent with the larger exposure of hydrophobic surfaces in the case of A-like sugars. Overall, the A-like nucleotides aid in achieving protein-induced fit in two major ways. First, hydrophobic clusters formed by several consecutive A-like sugars interact cooperatively with the non-polar surfaces in proteins. Second, the sugar switching occurs in large kinks promoted by direct protein contact, predominantly at the pyrimidine-purine dimeric steps. The sequence preference for the B-to-A sugar repuckering, observed for pyrimidines, suggests that the described DNA deformations contribute to specificity of the protein-DNA recognition in the minor groove.

Crystallography, X-Ray↗