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PubMed2005

Short-term dynamics of slurry-derived plant and microbial sugars in a temperate grassland soil as assessed by compound-specific delta13C analyses.

In view of recent discussions about climate change and the anthropogenically enhanced greenhouse effect, the aim of this study was to determine the short-term carbon (C) dynamics in a grassland soil after slurry application. It is known that, depending on cultivation practices, agro-ecosystems can act either as sources or as sinks for atmospheric CO2. C3 and C4 slurries were applied, differing in their stable C isotope signature, to be able to differentiate between native (soil-inherent) and fresh (slurry-applied) C. Samples were taken from 0-2, 2-7.5 and 7.5-15 cm soil depths from 90 days before until 4 weeks after slurry application at various intervals. We carried out compound-specific stable isotope analysis (CSIA) of plant- (arabinose and xylose) and microbial-derived sugars (fucose and rhamnose). Up to 45% of the applied slurry-derived xylose was found in the 0-2 cm soil depth within 24 h after slurry application, with this figure decreasing rapidly and then increasing again towards the end of the experiment. Therefore, during the first phase of slurry incorporation, preferentially the soluble part of slurry entered the first 2 cm of soil while, after about 2 weeks, particulate slurry-derived organic matter was incorporated into the soil. The ratio between plant- and microbial-derived sugars together with delta13C values of individual sugars in the 2-7.5 cm soil depth revealed that the dissipation of sugars from the 0-2 cm soil depth was not only due to leaching, but also was caused by microbial degradation of the fresh C because slurry did not contain significant amounts of rhamnose while the delta13C values of rhamnose became progressively enriched in 13C during the experiment. Stable isotope measurements of bulk soil previously only showed significant differences between C4 and C3 plots at 0-2 cm soil depth. The CSIA of the individual sugars was much more sensitive than bulk isotope measurements, revealing significant differences between C4 and C3 plots even at the 2-7.5 cm soil depth during the first phase of the experiment. Additionally, the dynamics of slurry-derived plant and microbial sugars could be followed specifically.

Bacteria↗

PubMed1996

Preparation of neoglycolipids with a definite sugar chain moiety from glycoproteins.

A method for preparing neoglycolipids possessing a sugar chain with a definite structure using two N-linked sugar chains as model compounds was developed. Pyridylaminated sugar chains obtained from glycoproteins were purified by reversed-phase HPLC and a pyridylaminated sugar chain thus obtained was converted to a 1-amino-1-deoxy derivative. The product was conjugated by reductive amination with phosphatidylglycolaldehyde prepared by periodate oxidation of dipalmitoyl phosphatidylglycerol. Neoglycolipids formed were purified by gel filtration from the reaction mixture. The structures of the purified neoglycolipids were confirmed by composition analysis for sugar, fatty acid, phosphorus, and 2-acetamido-1,2-dideoxy-1-[(hydroxyethyl)amino]-D-glucitol, the linkage region of the sugar chain, and lipid moieties. The method provides a convenient means of preparing neoglycolipids having a definite structure.

Carbohydrates↗

PubMed1999

Sugars exert a major influence on the vitrification properties of ethylene glycol-based solutions and have low toxicity to embryos and oocytes.

A systematic approach was taken to assess the vitrification properties of ethylene glycol-based solutions supplemented with carbohydrates. Solutions were prepared by weight (gravimetrically) using ethylene glycol as the cryoprotectant, 0.9% NaCl in water, and six different sugars: d-glucose, d(-)-fructose, d-sorbitol, sucrose, d(+)-trehalose, and raffinose. Sugars were added on a molal basis (0. 1, 0.5, and 1 m). Characteristics of the solutions were measured during warming by differential scanning calorimetry using a cooling rate of 100 degrees C/min and a warming rate of 10 degrees C/min. In the absence of carbohydrates a 59 wt% EG-saline solution formed a stable glass. When EG was replaced by an equimolal concentration of glucose, fructose, or sorbitol (monosaccharides) at 0.1, 0.5, or 1.0 m there was no change in the total solute concentration at which vitrification occurred, but the glass transition (Tg) occurred at a higher temperature than in EG-saline alone. When EG was replaced by an equimolal concentration of sucrose or trehalose (disaccharides) both the Tg and the lowest total solute concentration required for vitrification became progressively higher as the molecular weight, or the ratio of sugar to EG in the solutions, increased. At the highest tested disaccharide concentration (1 m) vitrification was achieved at a total solute concentration of 65 wt% (sucrose) and 67 wt% (trehalose). The polysaccharide raffinose significantly modified the vitrification properties of ethylene glycol solutions. When 0.5 or 0.1 m raffinose replaced EG on an equimolal basis the glass transition point was raised more than with either the monosaccharides or the disaccharides. Raffinose allowed vitrification at a total solute concentration of 67 wt% (0.5 m) and 63 wt% (0.1 m). The maturation of immature mouse oocytes, and the development of embryos in media containing 5-7 mM of any sugar was comparable to controls, indicating that they are not toxic. Exposure of freshly collected GV or MII oocytes to sugar concentrations between 0.5 and 1.0 M, for up to 10 min had no significant effect on the proportion which subsequently formed two cells. We conclude that added sugars do contribute to a solutions overall vitrification properties, and their properties should be taken into consideration when vitrification solutions are being designed or modified.

Animals↗

PubMed2002

Increased expression of p21(WAF-1/CIP-1) in the lens epithelium of rat sugar cataract.

It has been demonstrated that exquisite regulation of the cell cycle between the activation and inhibition is crucial to maintain the transparency of the ocular lens. While it is generally recognized that the sugar cataract is accompanied by the enhanced proliferation of lens epithelial cells (LECs), it is unclear whether or not an inhibitory mechanism against the lens proliferation is involved, except for TGF-beta. In this study, the authors demonstrated the enhanced expression of p21(WAF-1/CIP-1), a potent inhibitor against cell cycle progression, and its specific temporal and regional expression profiles in the LECs during the development of sugar cataract. Sugar cataract was induced in 6-week-old male Sprague-Dawley rats by feeding them on a 50% galactose-rich diet, and then the expression patterns of p21(WAF-1/CIP-1) mRNA and protein with the advance of the sugar cataract were studied. Western blot analyses showed that p21(WAF-1/CIP-1) expression increased throughout the period of galactose exposure, up to 21 days. Also, a gradual increase in the number of p21(WAF-1/CIP-1) positive cells was observed immunohistochemically in the course of the galactose exposure. Interestingly, p21(WAF-1/CIP-1) was significantly expressed in the multi-layered epithelium, which was observed typically in the advanced cataract. Proliferating cell nuclear antigen (PCNA), an indicator of cell proliferation, was also positive in the most multi-layered epithelial cells. In addition, transient expression of PCNA mRNA and its protein was noticed throughout the lens epithelium in the course of the sugar cataract development. Prior to the elevation of p21(WAF-1/CIP-1) mRNA expression, PCNA mRNA expression increased greatly and reached a peak according to the semiquantitative analyses using either the real time reverse transcription-polymerase chain reaction (RT-PCR) or the Southern blot analyses. Based on these observations, it is possible that p21(WAF-1/CIP-1) is elevated and exerts its inhibitory action against the proliferating epithelial cells during the development of the sugar cataract.

Animals↗

PubMed1996

Airflow obstruction in chalkpowder and sugar workers.

Our aim was to investigate the relationship between suspected occupational hazards and airflow obstruction in industrial workers. The study was a cross-sectional survey of 314 male workers from a chalkpowder plant (n = 158) and from a sugar refinery (n = 156). Occupational exposure to chalkpowder and sugar dust was assessed by individual job classification. Outcome variables included respiratory symptoms and routine spirometric parameters. Statistical analysis was done for each industry separately. Overall, mean pulmonary function parameters fell either within or above the normal range in both industries. However, analysis by job classification showed that in the chalkpowder plant, all indices of airway obstruction declined significantly with increasing dustiness. Additionally, workers in the dustiest workplace (chalk sacking) had significantly lower airflow parameters than workers from other workstations. In the sugar refinery, workers exposed to sugar dust in the sugar cube manufacture workstation had significantly lower forced expiratory volume in 1 s (FEV1) (p = 0.02) than the non-exposed ones. For both industries, the proportion of subjects complaining of cough and/or phlegm was greater among the most exposed subgroups than among the remaining workers but the differences were not statistically significant. In conclusion, coupling spirometry to job classification proved useful in disclosing a relationship between airflow obstruction and exposure to either chalkpowder or sugar dust in industrial workers.

Adult↗

PubMed1987

The rise of blood sugar as an additional parameter in traumatic shock.

Besides the known cardiovascular effects of hemorrhagic shock, regular metabolic consequences can be demonstrated in the rabbit: 1. The rise in blood sugar in hemorrhagic shock increases with the amount of blood withdrawn per unit of time. 2. The characteristic rise in blood sugar lasts longer than the phase of the partial exsanguination. 3. In traumatic shock, the rise in blood sugar indicates the degree of severity of the injury earlier than the fall in blood pressure does. The rise in blood sugar is attributable to a reflex increase in glycogenolysis due to catecholamine secretion elicited via baroreceptors in hemorrhagic shock. For the pronounced rise in blood sugar in traumatic shock, an additional receptor is postulated. Supplementary investigation of the blood sugar may possibly enable the extent of the injury to be diagnosed earlier in traumatic shock, and the negative sequelae may be reduced by timely, adequate therapy.

Animals↗

PubMed1992

Asparagine-linked sugar chains of erythrocyte membrane glycoproteins from Wiskott-Aldrich syndrome are not impaired.

In both healthy controls and patients with Wiskott-Aldrich syndrome, the main oligosaccharide in asparagine-linked sugar chains of the membrane glycoproteins of erythrocytes was biantennary sugar chain with bisected GlcNAc (Gal2-GlcNAc2-Man3-GlcNAc-GlcNAc-Fuc-GlcNAcOT). Biantennary sugar chain with an alpha-fucosyl residue linked at the proximal GlcNAc was seen but biantennary sugar chain without an alpha-fucosyl residue at the proximal GlcNAc was not detected in each subject. There was no difference in quality and quantity of asparagine-linked sugar chains of erythrocyte membrane glycoproteins between healthy controls and the patients. These results suggest that asparagine-linked sugar chains in membrane glycoproteins of hematopoietic cells may not be impaired in Wiskott-Aldrich syndrome.

Adolescent↗

PubMed1992

Asparagine-linked sugar chains of plasma membrane glycoproteins from healthy and common variable immunodeficiency B lymphoblastoid cell lines.

Asparagine-linked sugar chains of plasma membrane glycoproteins, which are formed by glycosylation during B cell maturation, were examined with B lymphoblastoid cell lines (LCLs) transformed by Epstein-Barr virus derived from healthy controls and patients with common variable immunodeficiency (CVI). Both two patients with CVI showed hypogammaglobulinemia and impaired B cell functions. LCLs from healthy controls and the patients showed CD19+ and HLA/DR+ in the cell surface and secreted IgM. In both healthy controls and the patients, the main oligosaccharide in asparagine-linked sugar chains of the membrane glycoproteins of LCLs was biantennary sugar chain with bisected GlcNAc (Gal2-GlcNAc2-Man3-GlcNAc-GlcNAc-Fuc-GlcNAcOT). Biantennary sugar chain with an alpha-fucosyl residue linked at the proximal GlcNAc was seen but biantennary sugar chain without an alpha-fucosyl residue at the proximal GlcNAc was little detected in each LCL. There was no difference in quality and quantity of asparagine-linked sugar chains between healthy controls and the patients. These results suggest that glycosylation during B cell maturation may not be impaired in patients with CVI.

Asparagine↗

PubMed1984

Behavior of sugar derivatives in procedures for ganglioside isolation.

A common method of studying ganglioside metabolism is to measure the amounts of radioactivity incorporated into ganglioside from a radiolabeled precursor. This requires that radioactive nonganglioside material be completely removed from the ganglioside fraction. Nucleotide sugars and aminosugars comprise an important source of such contaminants. Therefore, we have studied their behaviors in several procedures currently employed to isolate gangliosides. Over 50% of the radioactivity associated with several nucleotide sugars added to a brain homogenate is extracted with chloroform/methanol (2:1, v/v), and most of this is recovered in the upper phase of a Folch partition. Dialysis against water removes almost all of the free aminosugar but only 70% of nucleotide sugar. Treatment with alkaline phosphatase, phosphodiesterase and alkaline methanol followed by dialysis removes almost all of the nucleotide diphosphate sugars but only 88% of cytidine 5'-monophosphate sialic acid (CMP-NeuAc). Nucleotide sugars cannot be separated from gangliosides by Unisil or Iatrobead chromatography, but nucleotide diphosphate sugars and gangliosides are resolved with Sephadex LH-20 chromatography following treatment with phosphodiesterase and alkaline phosphatase. CMP-NeuAc was not satisfactorily separated from gangliosides using any of the procedures.

Brain Chemistry↗

PubMed2007

Phosphate starvation responses are mediated by sugar signaling in Arabidopsis.

Phosphate (Pi) is one of the least available plant nutrients in soils. It is associated with dynamic changes in carbon fluxes and several crucial processes that regulate plant growth and development. Pi levels regulate the expression of large number of genes including those involved in photosynthesis and carbon metabolism. Herein we show that sugar is required for Pi starvation responses including changes in root architecture and expression of phosphate starvation induced (PSI) genes in Arabidopsis. Active photosynthesis or the supplementation of sugar in the medium was essential for the expression of PSI genes under Pi limiting conditions. Expression of these genes was not only induced by sucrose but also detected, albeit at reduced levels, with other metabolizable sugars. Non-metabolizable sugar analogs did not induce the expression of PSI genes. Although sugar input appears to be down-stream of initial Pi sensing, it is absolutely required for the completion of the PSI signaling pathway. Altered expression of PSI genes in the hexokinase signaling mutant gin2 indicates that hexokinase-dependent signaling is involved in this process. The study provides evidence for requirement of sugars in PSI signaling and evokes a role for hexokinase in some components of Pi response mechanism.

Arabidopsis↗

PubMed2006

Role of sugars in regulating transfer cell development in cotyledons of developing Vicia faba seeds.

Transfer cell formation in cotyledons of developing faba bean (Vicia faba L.) seeds coincides with an abrupt change in seed apoplasm composition from one dominated by hexoses to one in which sucrose is the principal sugar. On the basis of these observations, we tested the hypothesis that sugars induce and/or sustain transfer cell development. To avoid confounding effects of in planta developmental programs, we exploited the finding that adaxial epidermal cells of cotyledons, which do not become transfer cells in planta, can be induced to form functional transfer cells when cotyledons are cultured on an agar medium. Growth rates of cotyledons cultured on hexose or sucrose media were used to inform choice of sugar concentrations. The same proportion of adaxial epidermal cells of excised cotyledons were induced to form wall ingrowths independent of sugar species and concentration supplied. In all cases, induction of wall ingrowths coincided with a marked increase in the intracellular sucrose-to-hexose ratio. In contrast, further progression of wall ingrowth deposition was correlated positively with intracellular sucrose concentrations that varied depending upon external sugar species and supply. Sucrose symporter induction and subsequent maintenance behaved identically to wall ingrowth formation in response to an external supply of hexoses or sucrose. However, in contrast to wall ingrowth formation, induction of sucrose symporter activity was delayed. We discuss the possibility of intracellular sugars functioning both as signals and substrates that induce and control subsequent development of transfer cells.

Carbohydrates↗

PubMed2004

An ethanologenic yeast exhibiting unusual metabolism in the fermentation of lignocellulosic hexose sugars.

Three lignocellulosic substrate mixtures [liquid fraction of acid-catalyzed steam-exploded softwood, softwood spent sulfite liquor (SSL) and hardwood SSL] were separately fermented by the industrially employed SSL-adapted strain Tembec T1 and a natural galactose-assimilating isolate (Y-1528) of Saccharomyces cerevisiae to compare fermentative efficacy. Both strains were confirmed as S. cerevisiae via molecular genotyping. The performance of strain Y-1528 exceeded that of Tembec T1 on all three substrate mixtures, with complete hexose sugar consumption ranging from 10 to 18 h for Y-1528, vs 24 to 28 h for T1. Furthermore, Y-1528 consumed galactose prior to glucose and mannose, in contrast to Tembec T1, which exhibited catabolite repression of galactose metabolism. Ethanol yields were comparable regardless of the substrate utilized. Strains T1 and Y-1528 were also combined in mixed culture to determine the effects of integrating their distinct metabolic capabilities during defined hexose sugar and SSL fermentations. Sugar consumption in the defined mixture was accelerated, with complete exhaustion of hexose sugars occurring in just over 6 h. Galactose was consumed first, followed by glucose and mannose. Ethanol yields were slightly reduced relative to pure cultures of Y-1528, but normal growth kinetics was not impeded. Sugar consumption in the SSLs was also accelerated, with complete utilization of softwood- and hardwood-derived hexose sugars occurring in 6 and 8 h, respectively. Catabolite repression was absent in both SSL fermentations.

Cellulose↗

PubMed2005

Phenolic compounds in red oak and sugar maple leaves have prooxidant activities in the midgut fluids of Malacosoma disstria and Orgyia leucostigma caterpillars.

Phenolic compounds are generally believed to be key components of the oxidative defenses of plants against pathogens and herbivores. However, phenolic oxidation in the gut fluids of insect herbivores has rarely been demonstrated, and some phenolics could act as antioxidants rather than prooxidants. We compared the overall activities of the phenolic compounds in red oak (Quercus rubra) and sugar maple (Acer saccharum) leaves in the midgut fluids of two caterpillar species, Malacosoma disstria (phenolic-sensitive) and Orgyia leucostigma (phenolic-tolerant). Three hypotheses were examined: (1) ingested sugar maple leaves produce higher levels of semiquinone radicals (from phenolic oxidation) in caterpillar midgut fluids than do red oak leaves; (2) O. leucostigma maintains lower levels of phenolic oxidation in its midgut fluids than does M. disstria; and (3) phenolic compounds in tree leaves have overall prooxidant activities in the midgut fluids of caterpillars. Sugar maple leaves had significantly lower ascorbate: phenolic ratios than did red oak leaves, suggesting that phenolics in maple would oxidize more readily than those in oak. As expected, semiquinone radicals were at higher steady-state levels in the midgut fluids of both caterpillar species when they fed on sugar maple than on red oak, consistent with the first hypothesis. Higher semiquinone radical levels were also found in M. disstria than in O. leucostigma, consistent with the second hypothesis. Finally, semiquinone radical formation was positively associated with two markers of oxidation (protein carbonyls and total peroxides). These results suggest that the complex mixtures of phenolics in red oak and sugar maple leaves have overall prooxidant activities in the midgut fluids of M. disstria and O. leucostigma caterpillars. We conclude that the oxidative defenses of trees vary substantially between species, with those in sugar maple leaves being especially active, even in phenolic-tolerant herbivore species.

Acer↗

PubMed2004

Identification and characterization of a novel cassava (Manihot esculenta Crantz) clone with high free sugar content and novel starch.

This study reports the identification of a new class of cassava (Manihot esculenta Crantz) with a storage root showing unusual free sugar accumulation and novel starch. Twenty-seven clones high in free sugar were identified under cultivation in primitive rural community areas in the Amazon. Iodine test and glucose oxidase-peroxidase reagent strips were used, in the field, for identification of starch and glucose, respectively. Five out of these 27 clones of cassava were cultivated at EMBRAPA Genetic Resources and Biotechnology and used for biochemical characterization, starch synthesis enzyme activities and gene expression analysis. Carbohydrates were fractioned into free sugar, polymerized water-soluble and -insoluble alpha-polyglucan. Clones of series CAS36 accumulate over 100 times more free sugar (mainly glucose) than commercial varieties. Monosaccharide composition analysis revealed one clone with distinct water-soluble sugars not present in the commercial cultivar. Structure analysis of the water-soluble and -insoluble alpha-polyglucan revealed the presence of a glycogen-like starch in clone CAS36.1. This clone indicated disruption in the starch synthesis pathway for enzyme activities and protein blot analyses in ADPG-pyrophosphorylase and branching enzyme, and their corresponding protein. Gene expression analysis indicated the lack of transcript for the gene coding for branching enzyme, but not for the gene coding for the ADPG-pyrophosphorylase small subunit. In addition, the pattern of distribution of sugar and starch content showed to be related to tissue age in the storage root.

1,4-alpha-Glucan Branching Enzyme↗

PubMed1985

Calcium-dependence of sugar transport in rat small intestine.

The involvement of Ca2+ in the theophylline action on sugar transport was investigated in isolated rat small intestinal mucosa. Theophylline significantly increased cell water free sugar accumulation and reduced mucosal to serosal sugar fluxes both in the presence and absence of calcium, but the effects of theophylline were significantly less in calcium free media. In theophylline untreated tissues, calcium-deprived bathing solutions decreased tissue galactose accumulation and increased mucosal to serosal sugar flux. The calcium-channel blocker verapamil produced similar effects on intestinal galactose transport to those induced by low extracellular calcium activity. RMI 12330A and the calmodulin antagonist trifluoperazine abolished the theophylline-effects on intestinal galactose transport. Both drugs also affected sugar transport in basal conditions. These studies suggest that calcium might modulate sugar permeability across the basolateral boundary of rat enterocytes, and that its effect may be mediated by calmodulin.

Animals↗

PubMed1984

Structural comparison of the two distinct sugar binding sites in wheat germ agglutinin isolectin II.

The two unique sugar binding sites in wheat germ agglutinin, located in the subunit/subunit interface of the dimer molecule and termed primary and secondary binding sites, are compared in the light of the newly obtained chemical amino acid sequence and a high-resolution electron density map (1.8 A). Homology was found in the three amino acid residues directly involved in sugar binding: Tyr73II, Ser62II, Glu115I in the primary site, and Tyr159I, Ser148I, Asp29II in the secondary site (subscripts refer to promoters I and II). Thirteen corresponding side-chain atoms of these three homologous residues in the two sites could be superimposed with a root-mean-square difference of 1.39 A. The three sugar binding residues are located in subsite 1 of each extended binding location and contribute to binding of the terminal, non-reducing N-acetyl-D-glucosamine and N-acetyl-D-neuraminic acid residues only, and they provide three hydrogen bonds for complex stabilization. Two hydrogen bonds are made with the carbonyl and amido portions of the N-acetyl group and the third with the C-3 OH group of the sugar ring. It is suggested that small differences in the sugar binding affinities at these two unique sites exist, due to the different numbers of van der Waals' interactions made at these sites, which contribute to stabilizing, for instance, the wheat germ agglutinin/N,N'-diacetyl-chitobiose complex. The single tryptophan residue is located at a distance of approximately 13 A from the primary site and is thought to have no affect on sugar binding. In addition, the disposition of the four saccharide binding sites of the dimer with respect to three local, pseudo 2-fold symmetry axes, relating domains of opposite protomers, is discussed.

Amino Acid Sequence↗

PubMed1992

A sensitive fluorometric assay for reducing sugars.

A simple and rapid fluorometric assay for reducing sugars that is sensitive to the nanomolar range has been developed. The assay involves the derivatization of a given sugar with hydrazine at pH 3 to form a hydrazone, which is reacted with fluorescamine following adjustment of pH to first 9.4 and then 7.4. The amount of sugar in a sample is quantitated by measuring the fluorescence intensity at an excitation wavelength of 400 nm and an emission wavelength of 490 nm. The assay is precise and reproducible, as indicated by intra- and inter-run variations of at most 3% and 4%, respectively. In addition to reducing sugars, the assay can also be used to measure aliphatic and aromatic aldehydes, but not acetone. Compared with an existing fluorometric sugar assay, the assay reported here does not require chromatographic separation of the fluorescent derivative from unreacted fluorescamine. The assay can, however, be potentially adapted for postcolumn detection of aldehydes, reducing sugars, and hydrazones in HPLC.

Aldehydes↗

PubMed1977

Studies on mutagenicity of irradiated sugar solutions in Salmonella typhimurium.

Irradiated sugar solutions are mutagenic towards Salmonella typhimurium, the effect being dose-dependent up to 2.0 Mrad. At all doses, ribose solution exhibited greater mutagenicity than did sucrose solution. The mutagenic effect was observed only in dividing cells and appears to be directly related to the growth rate. A larger proportion of revertants was observed after incubation with irradiated sugar solution for a period of 4 h than for 24 h. Irradiation of the sugar solutions in the frozen conditions was effective in completely preventing the development of mutagenic potential. Post-irradiation storage of the sugar solutions for a prolonged period (25 weeks) also minimized their mutagenic effect. The irradiated sugar solutions gave rise to both missense and frame-shift (additon as well as deletion) types of mutation; ribose was more effective in inducing the latter type. The irradiated sugar solutions failed to show a mutagenic response in the host-mediated assay with mice as the mammalian host.

Anaerobiosis↗