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Formation of PhIP in a mixture of creatinine, phenylalanine and sugar or aldehyde by aqueous heating.

A mixture of 100 mM creatinine and 100 mM L-phenylalanine was heated at 60 or 37 degrees C in the presence of sugar or aldehyde. A mutagen, 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) formed in the model system was determined by reversed-phase HPLC. Any sugars tested induced the formation of PhIP when heated at 60 degrees C, though PhIP was not detected in a mixture without sugar. Among the sugars tested, D-erythrose and D-glyceraldehyde were more productive than pentose (D-arabinose and D-ribose) and hexose (D-glucose and D-galactose) in the yield of PhIP. Moreover, PhIP was formed even when a mixture of creatinine, L-phenylalanine and D-glucose or D-ribose was incubated at 37 degrees C for a long time. Both formaldehyde and acetaldehyde also induced the formation of PhIP, though PhIP was not detected in a mixture without sugar or aldehyde even when heated at 100 degrees C. These results indicate that PhIP can be formed at low-temperature heating and that either sugar or aldehyde is essential for PhIP formation in the model system. Our data also suggest that aldehydes may be a key reactant in the formation of PhIP in aqueous heating of the mixture of creatinine and L-phenylalanine.

Acetaldehyde↗

The distribution of sugar chains on the vomeronasal epithelium observed with an atomic force microscope.

The distribution of sugar chains on tissue sections of the rat vomeronasal epithelium, and the adhesive force between the sugar and its specific lectin were examined with an atomic force microscope (AFM). AFM tips were modified with a lectin, Vicia villosa agglutinin, which recognizes terminal N-acetyl-D-galactosamine (GalNAc). When a modified tip scanned the luminal surface of the sensory epithelium, adhesive interactions between the tip and the sample surface were observed. The final rupture force was calculated to be approximately 50 pN based on the spring constant of the AFM cantilever. Distribution patterns of sugar chains obtained from the force mapping image were very similar to those observed using fluorescence-labeled lectin staining. AFM also revealed distribution patterns of sugar chains at a higher resolution than those obtained with fluorescence microscopy. Most of the adhesive interactions disappeared when the scanning solution contained 1 mM GaINAc. The adhesive interactions were restored by removing the sugar from the solution. Findings suggest that the adhesion force observed are related to the binding force between the lectin and the sugars distributed across the vomeronasal epithelium.

Acetylgalactosamine↗

Intrinsic nitric oxide regulates the taste response of the sugar receptor cell in the blowfly, Phormia regina.

The taste organ in insects is a hair-shaped taste sensory unit having four functionally differentiated contact chemoreceptor cells. In the blowfly, Phormia regina, cGMP has been suggested to be a second messenger for the sugar receptor cell. Generally, cGMP is produced by membranous or soluble guanylyl cyclase (sGC), which can be activated by nitric oxide (NO). In the present paper, we electrophysiologically showed that an NO scavenger, 2-phenyl-4,4,5,5-tetramethylimidazoline-3-oxide-1-oxyl (PTIO), an NO donor, 1-hydroxy-2-oxo-3-(N-methyl-3-aminopropyl)-3-methyl-1-triazene (NOC 7) or an NO synthase (NOS) inhibitor, NG-nitro-L-arginine methyl ester (L-NAME) specifically affected the response in the sugar receptor cell, but not in other receptor cells. PTIO, when introduced into the receptor cells in a sensillum aided by sodium deoxycholate (DOC, pH 7.2), depressed the response of sugar receptor cells to sucrose but did not affect those of the salt or water receptor cells. NOC 7, given extracellularly, latently induced the response of sugar receptor cells; and L-NAME, when introduced into the receptor cells, depressed the response of sugar receptor cells. The results clearly suggest that NO, which may be produced by intrinsic NOS in sugar receptor cells, participates in the transduction cascade of these cells in blowfly.

Animals↗

Developmental changes in sugar responses of the chorda tympani nerve in preweanling rats.

To clarify developmental changes in the gustatory system of the rat, integrated taste responses from the chorda tympani (CT) nerve were recorded and analyzed at different postnatal ages. The response magnitude was calculated relative to the response to the standard, 0.1 M NH4Cl. Even at 1 week of age, the CT responded well to all tested 0.1 M chloride salts (NH4Cl, NaCl, LiCl, KCl, RbCl and CsCl). The responses to 0.1 M NaCl and LiCl increased with increasing age of the rat while response magnitudes to KCl, RbCl and CsCl did not change up to 8 weeks. At 1 week, the integrated response pattern was quite similar to that in adult rats for NaCl, HCl and quinine hydrochloride (QHCl). The concentration-response functions for NaCl, HCl, QHCl and sucrose at 2 weeks were essentially the same as those at 8 weeks. These results suggest that taste buds in the 2-week-old rat are functionally mature for the detection of the four basic taste stimuli. The relative magnitude of the responses to the various sugars was smaller at 1 week compared to the adult rat and reached a maximum at weeks 3-4, then decreased gradually with age. Among the six sugars, sucrose was the most effective followed by lactose. From weeks 1-4, the magnitude of the integrated taste response to fructose was smaller than that to lactose except at 3 weeks of age. Maltose, galactose and glucose were less potent stimuli than the other sugars tested. The response magnitude to lactose at 4 weeks had decreased compared to that for the other sugars. Taste responses to the sugars in preweanling and adult rats were not cross-adapted by the individual sugars. These results suggest that after 1 week of age during postnatal development in the rat, taste information from the CT rapidly increases in its importance for feeding behavior.

Aging↗

Semiquantitative determination of short-chain fatty acids in cane and beet sugars.

Some sugars, specifically white beet sugar and raw cane sugars, possess off-flavors and off-odors. Although not necessarily the source, the presence of short-chain fatty acids serves as an indicator of an off-odor problem in sugar. Solid-phase microextraction (SPME) is used to collect the volatile compounds from the headspace of sugar. The temperature, moisture, and type of SPME fiber are varied to optimize recovery. Sugars analyzed in the absence of water using an incubation temperature of 70 degrees C with a divinylbenzene-carboxen-polydimethylsiloxane fiber yield the most reproducible results. Data from depletion analyses report a recovery level of 38% for the first injection. The semiquantitative analysis of butyric acid is accomplished using injected standards to develop a calibration curve.

Calibration↗

Structures and functional roles of the sugar chains of human erythropoietins.

Erythropoietin (EPO) is a haemopoietic hormone specific to cells of erythroid lineage. EPO has recently become available for the treatment of anaemia as the first human recombinant biomedicine produced in heterologous mammalian cells. Human EPO is characterized by its large carbohydrate chains, which occupy close to 40% of its total mass. These sugar moieties were thought to be important for the biological activity of EPO, but detailed studies were not performed until the structures were elucidated. The variety of roles for the sugar chains were then immediately found once the structures were known. EPO is an excellent model for investigating the roles of sugar chains on glycoproteins, since its gene and its multiple glycoforms are available, as well as sensitive bioassays for testing. In this review, we will first summarize the known sugar chain structures of EPO from different host cells, and then discuss the host-cell dependent and peptide structure-dependent glycosylation of glycoproteins. We will then address how one investigates the roles of sugar chains of glycoproteins, show several examples of such investigations, and discuss the functional roles of HuEPO's sugar chains in its biosynthesis and secretion, its in vitro and in vivo biological activities, and its half-life in blood circulation.

Carbohydrate Sequence↗

Photosynthesis, sugars and the regulation of gene expression.

Sugar-mediated regulation of gene expression is a mechanism controlling the expression of many different plant genes. In this review, a compilation of the genes encoding photosynthetic proteins, subject to this mode of regulation, is presented. Several groups have devised different screening strategies to obtain Arabidopsis mutants in sugar sensing and signalling. An overview of these strategies has been included. Sugar-mediated regulation of gene expression is thought to require the hexokinase (HXK) protein. It has previously been shown that one such sugar, mannose, is capable of blocking germination in Arabidopsis. This inhibition is also mediated by HXK and occurs in the low millimolar concentration range. Here, the use of germination on mannose as an effective screening strategy for putative sugar sensing and signalling mutants is reported. T-DNA- and EMS-mutagenized collections were used to isolate 31 mannose-insensitive germination (mig) mutants. With the use of these mutants, a comparison between this screen and other existing sugar-sensing screens is presented.

Arabidopsis↗

Female Aedes aegypti (Diptera: Culicidae) in Thailand rarely feed on sugar.

Female Aedes aegypti (L.) of two different body sizes and provided with different diets (20% sucrose, water only, or 20% sucrose + human blood) were marked and released together in a rural Thai village. Recaptured marked and unmarked (wild) adults were tested for fructose by the cold-anthrone reaction. Both released and wild females showed a low frequency of sugar feeding in nature; both small-bodied and large-bodied individuals failed to sugar feed significantly. Marked females released with sugar in their crop utilized this energy source over the following 2-3 d but failed to replenish it. In contrast, about one-third of wild, resting males showed evidence of recent sugar feeding. This indicated that the absence of plant sugar in females was not caused by a shortage of nectar sources in this rural domestic environment. Presumably, the nonutilization of plant sugar as an energy source increases the frequency of blood feeding and, therefore, the vectorial capacity of Ae. aegypti.

Aedes↗

Spermidine and related-metabolic inhibitors modulate sugar and amino acid levels in Vitis vinifera L.: possible relationships with initial fruitlet abscission.

The relationships between free polyamines (PAs), sugar and amino acid status were investigated in cuttings from two cultivars of Vitis vinifera L., Pinot noir (PN), a low abscising cultivar and Merlot (MRT), a high abscising one. In both cultivars free PAs decreased in inflorescences, but more drastically in MRT plants. Upon anthesis, this was associated with a decreased sugar content, especially sucrose, and an increase in total free amino acids. Thereafter, sucrose and amino acids showed opposite trends. In addition, darkening the PN plants at full flowering resulted in a dramatic decrease of PAs and sugars in inflorescences, but an increase in amino acid content, followed by high abscission. The concept that initial fruitlet abscission might be regulated by free PAs through changes in primary metabolites was hypothesized. Hence, the application of exogenous spermidine (Spd), but not putrescine (Put), prior to flowering markedly inhibits abscission. The Spd treatment also increased soluble sugar content but reduced amino acids in both leaves and inflorescences, while Put had no significant effect. By contrast, cyclohexylamine and beta-hydroxyethylhydrazine, as potent inhibitors of Spd synthase and PA-oxidases, respectively, exerted inverse effects on sugar, amino acid and abscission levels. Sucrose and free proline seemed to be highly sensitive to these treatments. This study suggests that Spd could regulate fruitlet abscission in grapevine by modulating, in a reverse way, the levels of sugars and amino acids in inflorescences.

Amino Acids↗

Sugar and phytohormone response pathways: navigating a signalling network.

Many plant developmental, physiological and metabolic processes are regulated, at least in part, by nutrient availability. In particular, alterations in the availability of soluble sugars, such as glucose and sucrose, help regulate a diverse array of processes. Multiple lines of evidence indicate that many of these processes are also regulated in response to other signalling molecules, such as phytohormones. This review draws examples from a variety of plant systems, including bean, Arabidopsis, potato, and cereals. Five of the most interesting and best developed examples of processes regulated via 'interactions' or 'crosstalk' between sugars and phytohormones are described, including embryogenesis, seed germination, early seedling development, tuberization, and the regulation of alpha-amylase activity. The types of mechanisms by which different response pathways are known or postulated to interact are also described. These mechanisms include regulation of the metabolism and/or transport of a signalling molecule by a different response pathway. For example, sugars have been postulated to help regulate the synthesis, conjugation and/or transport of phytohormones, such as gibberellins and abscisic acid. Conversely, phytohormones, such as abscisic acid, gibberellins and cytokinins have been shown to help regulate sugar metabolism and/or transport. Similarly, sugars have been shown to regulate the expression of components of phytohormone-response pathways and phytohormones regulate the expression of some genes encoding possible components of sugar-response pathways. Examples of proteins and second messengers that appear to act in multiple response pathways are also described.

Arabidopsis↗

UVA-visible photo-excitation of guanine radical cations produces sugar radicals in DNA and model structures.

This work presents evidence that photo-excitation of guanine radical cations results in high yields of deoxyribose sugar radicals in DNA, guanine deoxyribonucleosides and deoxyribonucleotides. In dsDNA at low temperatures, formation of C1'* is observed from photo-excitation of G*+ in the 310-480 nm range with no C1'* formation observed > or =520 nm. Illumination of guanine radical cations in 2'dG, 3'-dGMP and 5'-dGMP in aqueous LiCl glasses at 143 K is found to result in remarkably high yields (approximately 85-95%) of sugar radicals, namely C1'*, C3'* and C5'*. The amount of each of the sugar radicals formed varies dramatically with compound structure and temperature of illumination. Radical assignments were confirmed using selective deuteration at C5' or C3' in 2'-dG and at C8 in all the guanine nucleosides/tides. Studies of the effect of temperature, pH, and wavelength of excitation provide important information about the mechanism of formation of these sugar radicals. Time-dependent density functional theory calculations verify that specific excited states in G*+ show considerable hole delocalization into the sugar structure, in accord with our proposed mechanism of action, namely deprotonation from the sugar moiety of the excited molecular radical cation.

Cations↗

Structural analysis of N-linked sugar chains of human blood clotting factor IX.

The structures of N-glycans of human blood clotting factor IX were studied. N-Glycans liberated by hydrazinolysis were N-acetylated and the reducing-end sugar residues were tagged with 2-aminopyridine. The pyridylamino (PA-) sugar chains thus obtained were purified by HPLC. Each PA-sugar chain was analyzed by two-dimensional sugar mapping combined with glycosidase digestion. The major structures of the N-linked sugar chains of human factor IX were found to be sialotetraantennary and sialotriantennary chains with or without fucose residues. These highly sialylated sugar chains are located on the activation peptide of the protein.

Carbohydrate Sequence↗

Activation of the promoters of Arabidopsis genes for the branched-chain alpha-keto acid dehydrogenase complex in transgenic tobacco BY-2 cells under sugar starvation.

Sugar starvation exerted by sub-10 mM levels of sucrose on Arabidopsis T87 suspension-cultured cells triggered marked accumulation of the transcripts of genes for E1beta and E2 subunit of the branched-chain alpha-keto acid dehydrogenase complex. Similar levels of sugar starvation increased the luciferase activity in transgenic tobacco BY-2 lines expressing the Arabidopsis E1beta- or E2-promoter-luciferase fusion gene. These results indicate that sugar levels tightly regulate the E1beta and E2 promoter activity in the heterologous plant system. We further showed in the transgenic tobacco BY-2 lines that sugar-starvation-induced activation of the E1beta and E2 promoters was prevented by K-252a, an inhibitor of Ser/Thr protein kinase, and was enhanced by okadaic acid, an inhibitor of protein phosphatases. By contrast, the cauliflower mosaic virus 35S promoter activity in sugar-starved BY-2 cells was not significantly affected by K-252a and only slightly enhanced by okadaic acid. Taken together, we propose that transcriptional activation of genes for the branched-chain alpha-keto acid dehydrogenase complex and its modulation by specific protein kinases/phosphatases are of critical importance in branched-chain amino acid catabolism in plant cells under sugar starvation.

3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide)↗

Sugar retrieval by coats of developing seeds of Phaseolus vulgaris L. and Vicia faba L.

Influxes of glucose, fructose and sucrose were characterised for coat cells of developing seeds of Phaseolus vulgaris L. and Vicia faba L. by monitoring uptake of [(14)C]sugars into excised seed-coat halves and two different protoplast populations derived from seed coats. Sugar influxes by the two populations of protoplasts were similar for each sugar species [sucrose > (fructose approximately glucose)] and hexoses competed with sucrose. Concentration-dependent influxes of all three sugars by excised seed coats could be described by a simple directly proportional relationship between concentration ([S]) and uptake rate (v) in the physiological range of sugar concentrations (v approximately A.[S]). Alternatively, with the exception of fructose influx by Vicia, all could be fitted to a Michaelis-Menten relationship, as could sucrose uptake by Vicia protoplasts. Apparent K(m) values were high ( approximately 100-500 mM) compared with those reported for other systems. Sucrose transport was distinct from glucose and fructose transport in both species. Sugar influx was decreased by p-chloromercuribenzenesulfonic acid, carbonylcyanide m-chlorophenylhydrazone and erythrosin B. These responses are consistent with sugar/H(+) symport acting to retrieve photoassimilates leaked to the apoplasm during post-sieve element transport within seed coats.

4-Chloromercuribenzenesulfonate↗

Involvement of HLS1 in sugar and auxin signaling in Arabidopsis leaves.

Sugar regulates a variety of genes and controls plant growth and development similarly to phytohormones. As part of a screen for Arabidopsis mutants with defects in sugar-responsive gene expression, we identified a loss-of-function mutation in the HOOKLESS1 (HLS1) gene. HLS1 was originally identified to regulate apical hook formation of dark-grown seedlings (Lehman et al., 1996, Cell 85: 183-194). In hls1, sugar-induced gene expression in excised leaf petioles was more sensitive to exogenous sucrose than that in the wild type. Exogenous IAA partially repressed sugar-induced gene expression and concomitantly activated some auxin response genes such as AUR3 encoding GH3-like protein. The repression and the induction of gene expression by auxin were attenuated and enhanced, respectively, by the hls1 mutation. These results suggest that HLS1 plays a negative role in sugar and auxin signaling. Because AUR3 GH3-like protein conjugates free IAA to amino acids (Staswick et al., 2002, Plant Cell 14: 1405-1415; Staswick et al., 2005, Plant Cell 17: 616-627), enhanced expression of GH3-like genes would result in a decrease in the free IAA level. Indeed, hls1 leaves accumulated a reduced level of free IAA, suggesting that HLS1 may be involved in negative feedback regulation of IAA homeostasis through the control of GH3-like genes. We discuss the possible mechanisms by which HLS1 is involved in auxin signaling for sugar- and auxin-responsive gene expression and in IAA homeostasis.

Arabidopsis↗

Relationship between temperature, respiratory loss of sugar and premature dehardening in dormant Scots pine seedlings.

Increased intracellular sugar concentration is an important contributor to the increased cold tolerance of conifers in winter. This study examines the extent to which wintertime respiratory loss of sugars leads to premature dehardening. Two-year-old seedlings of Scots pine (Pinus sylvestris L.), grown and cold-hardened in the field, were exposed to different temperature regimes for 16 weeks while dormant. To minimize short-term carry-over effects, after the temperature treatments, all seedlings were conditioned to 5.5 degrees C and watered before the assessment of non-structural carbohydrates and cold tolerance. Needle sugar concentration was decreased by 54, 32, 21 and 9% following treatment at 5.5, 0, -1.5 and -8.5 degrees C, respectively. Sugar concentration did not decrease as much in root tissues as in needles because starch was mobilized in roots. Cold tolerance of needles was analyzed by controlled freezing, and the temperature causing an initial 10% damage (LT(10)) was plotted as a function of needle sugar concentration, revealing a strong, linear relationship. When one-third of the initial sugars had been consumed, LT(10) had increased from -24.5 to -16.5 degrees C, and when one half had been consumed, LT(10) had increased to -10 degrees C. Consequences of these findings for the field performance of conifers are discussed in relation to climatic variation and change.

Journal Article↗

Chemotaxis towards sugars by Bacillus subtilis.

Many sugars and derivatives were tested in the capillary assay for their attraction of Bacillus subtilis. The major attractants were 2-deoxy-D-glucose, D-fructose, gentiobiose, D-glucose, maltose, D-mannitol, D-mannose, N-acetylglucosamine, alpha-methyl-D-glucoside, beta-methyl-D-glucoside, N-acetylmannosamine, alpha-methyl-D-mannoside, D-sorbitol, L-sorbose, sucrose, trehalose and D-xylose. Only glucose chemotaxis was completely constitutive. Competition experiments were carried out to determine the specificities of chemoreceptors. There were 25 instances of no influence of two sugars on each other's taxis, 92 instances of one sugar interfering non-reciprocally with chemotaxis towards another and 49 instances of two sugars reciprocally competing. However, in most of the last instances, other sugars were identified that interfered with chemotaxis towards one member of the pair but not the other. Thus, nearly all sugars and related compounds appear to be detected by their own chemoreceptors, but many secondary interactions exist.

Bacillus subtilis↗

QTL analysis in a complex autopolyploid: genetic control of sugar content in sugarcane.

QTL mapping in autopolyploids is complicated by the possibility of segregation for three or more alleles at a locus and by a lack of preferential pairing, however the subset of polymorphic alleles that show simplex segregation ratios can be used to locate QTLs. In autopolyploid Saccharum, 36 significant associations between variation in sugar content and unlinked loci detected by 31 different probes were found in two interspecific F(1) populations. Most QTL alleles showed phenotypic effects consistent with the parental phenotypes, but occasional transgressive QTLs revealed opportunities to purge unfavorable alleles from cultivars or introgress valuable alleles from exotics. Several QTLs on homologous chromosomes appeared to correspond to one another-multiple doses of favorable 'alleles' at such chromosomal region(s) yielded diminishing returns-such negative epistasis may contribute to phenotypic buffering. Fewer sugar content QTLs were discovered from the highest-sugar genotype than from lower-sugar genotypes, perhaps suggesting that many favorable alleles have been fixed by prior selection, i.e. that the genes for which allelic variants (QTLs) persist in improved sugarcanes may be a biased subset of the population of genes controlling sugar content. Comparison of these data to mutations and QTLs previously mapped in maize hinted that seed and biomass crops may share a partly-overlapping basis for genetic variation in carbohydrate deposition. However, many QTLs do not correspond to known candidate genes, suggesting that other approaches will be necessary to isolate the genetic determinants of high sugar content of vegetative tissues.

Alleles↗