Some carbohydrates found in pollen and pollen substitutes are toxic to honey bees

Carbohydrates in some pollen substitutes (galactose, lactose, raffinose, stachyose, glucuronic acid, galacturonic acid, polygalacturonic acid, and pectin) were toxic to caged adult Apis mellifera L. These toxins can be diluted to safe levels by sucrose. Collected nectar apparently dilutes the toxic sugars in pollen thus permitting assimilation of essential nutrients from pollen.     

Comparative quantitative analysis of sucrose and related compounds using ion exchange and reverse phase chromatographic methods

Two analytical methods of sugar determination, namely ion exchange chromatography on an anionic resin coupled with electrochemical detection, and reverse phase chromatography on Nucleosil-NH2 resin equipped with a light scattering detector were tested and compared as regards their rapidity, sensitivity and accuracy with sucrose, fructose, glucose, raffinose, maltose, arabinose, fucose, rhamnose and xylose. Excellent resolution and highly reproducible results were obtained in both cases. Greater sensitivity up to the picomolar range was possible however only with ion exchange chromatography. Reverse phase chromatography was successfully applied to the time course of sucrose hydrolysis under chemical (acid) and enzymatic (invertase) conditions. The hydrolysis was monitored by determining sucrose degradation and the corresponding formation of glucose and fructose.     

Lectin-like inhibition of immune complex receptor-mediated stimulation of neutrophils. Effects on cytosolic calcium release and superoxide production

We have tested the role of lectin-like interactions, with particular emphasis on CR3, in insoluble immune complex (IC)-mediated activation of human polymorphonuclear leukocytes (PMN). The ability of IC to trigger intracellular Ca2+ release and O2- production by normal PMN, saccharide-treated cells, and CR3-deficient PMN (leukocyte adhesion deficiency, LAD, patients) were tested. When indo-1-labeled normal PMN were stimulated with IC in Ca(2+)-free buffer, intracellular Ca2+ rose from approximately 100 nM to approximately 230 nM. However, when LAD PMN were tested, a small rise in intracellular Ca2+ was observed. Because previous studies have shown that certain saccharides inhibit CR3-Fc gamma RIII co-capping, we tested a panel of saccharides to determine their ability to influence IC-mediated intracellular Ca2+ release. When normal PMN were exposed to 0.15 M N-acetyl-D-glucosamine (NADG), D-mannose, or alpha-methyl-mannoside, the Ca2+ response to IC was significantly reduced. However, addition of 0.15 M glucose, raffinose, sucrose, galactose, fructose, or sorbitol did not significantly affect the Ca2+ response, suggesting that the response was specific for certain saccharides. To test the physiologic consequences of these Ca2+ signals, we have examined the ability of saccharides to influence O2- production by normal PMN and the ability of LAD PMN to produce O2- upon triggering by IC. Normal PMN stimulated with IC generated 4.3 +/- 1.7 nmol/10(6) cells/30 min of O2-. In contrast, O2- production was inhibited by 0 to 20% by glucose, galactose, sucrose, sorbitol, fructose, and raffinose and > or = 50% by NADG and mannose. LAD PMN, which display diminished Ca2+ signals, were found to produce O2- at 47 +/- 6% of control levels. NADG and mannose dose-response studies indicated that they cooperatively block O2-.     

Lactobacillus casei 64H contains a phosphoenolpyruvate-dependent phosphotransferase system for uptake of galactose, as confirmed by analysis of ptsH and different gal mutants

Galactose metabolism in Lactobacillus casei 64H was analyzed by genetic and biochemical methods. Mutants with defects in ptsH, galK, or the tagatose 6-phosphate pathway were isolated either by positive selection using 2-deoxyglucose or 2-deoxygalactose or by an enrichment procedure with streptozotocin. ptsH mutations abolish growth on lactose, cellobiose, N-acetylglucosamine, mannose, fructose, mannitol, glucitol, and ribitol, while growth on galactose continues at a reduced rate. Growth on galactose is also reduced, but not abolished, in galK mutants. A mutation in galK in combination with a mutation in the tagatose 6-phosphate pathway results in sensitivity to galactose and lactose, while a galK mutation in combination with a mutation in ptsH completely abolishes galactose metabolism. Transport assays, in vitro phosphorylation assays, and thin-layer chromatography of intermediates of galactose metabolism also indicate the functioning of a permease/Leloir pathway and a phosphoenolpyruvate-dependent phosphotransferase system (PTS)/tagatose 6-phosphate pathway. The galactose-PTS is induced by growth on either galactose or lactose, but the induction kinetics for the two substrates are different.     

Distinct regions of frequent loss of heterozygosity of chromosome 5p and 5q in human esophageal cancer

Twenty strains of Streptococcus bovis grew more slowly on lactose (1.21 +/- 0.12 h-1) then than on glucose (1.67 +/- 0.12 h-1), and repeated transfers or prolonged growth in continuous culture (more than 200 generations each) did not enhance the growth rate on lactose. Lactose transport activity was poorly correlated with growth rate, and slow growth could not be explained by the ATP production rate (catabolic rate). Batch cultures growing on lactose always had less intracellular fructose 1,6-bisphosphate (Frul,6P2) than cells growing on glucose (6.6 mM compared to 16.7 mM), and this difference could be explained by the pathway of carbon metabolism. Glucose and the glucose moiety of lactose were metabolized by the Embden-Meyerhoff-Parnas (EMP) pathway, but the galactose moiety of lactose was catabolized by the tagatose pathway, a scheme that by-passed Frul,6P2. A mutant capable of co-metabolizing lactose and glucose grew more rapidly when glucose was added, even though the total rate of hexose fermentation did not change. Wild-type S. bovis grew rapidly with galactose and melibiose, but these galactose-containing sugars were activated by galactokinase and catabolized via EMP. On the basis of these results, rapid glycolytic flux through the EMP pathway is needed for the rapid growth (more than 1.2 h-1) of S. bovis.     

Binding of monoclonal antibody 4B1 to homologs of the lactose permease of Escherichia coli

The conformationally sensitive epitope for monoclonal antibody (mAb) 4B1, which uncouples lactose from H+ translocation in the lactose permease of Escherichia coli, is localized in the periplasmic loop between helices VII and VIII (loop VII/VIII) on one face of a short helical segment (Sun J, et al., 1996, Biochemistry 35;990-998). Comparison of sequences in the region corresponding to loop VII/VIII in members of Cluster 5 of the Major Facilitator Superfamily (MFS), which includes five homologous oligosaccharide/H+ symporters, reveals interesting variations. 4B1 binds to the Citrobacter freundii lactose permease or E. coli raffinose permease with resultant inhibition of transport activity. Because E. coli raffinose permease contains a Pro residue at position 254 rather than Gly, it is unlikely that the mAb recognizes the peptide backbone at this position. Consistently, E. coli lactose permease with Pro in place of Gly254 also binds 4B1. In contrast, 4B1 binding is not observed with either Klebsiella pneumoniae lactose permease or E. coli sucrose permease. When the epitope is transferred from E. coli lactose permease (residues 245-259) to the sucrose permease, the modified protein binds 4B1, but the mAb has no significant effect on sucrose transport. The studies provide further evidence that the 4B1 epitope is restricted to loop VII/VIII, and that 4B1 binding induces a highly specific conformational change that uncouples substrate and H+ translocation.     

Trypanosoma evansi in capybara from Venezuela

Twenty-nine strains of yeasts, which are capable of using galactose, melibiose, or raffinose, were screened for alpha-galactosidase production. Among the strains, 5 produced intracellular and extracellular alpha-galactosidases, and 2 produced only intracellular enzyme. Substrate specificities of these enzymes were explored using 6(3)-alpha-D-galactosyl-1,4-beta-D-mannotriose and 6(3)-alpha-D-galactosyl-1,4-beta-D-mannotetraose. All enzymes liberated the terminal galactose from 6(3)-alpha-D-galactosyl-1,4-beta-D-mannotriose, but did not the stub galactose from 6(3)-alpha-D-galactosyl-1,4-beta-D-mannotetraose.     

Differential effects of sucrose, fructose, glucose, and lactose on crying in 1- to 3-day-old human infants: Qualitative and quantitative considerations.

Assessed the potency of different sugars as calming agents in human infants. Five 0.1-ml aliquots of 0.51M sucrose, fructose, glucose, or lactose were presented to 1- to 3-day-old infants who were crying spontaneously. Sucrose and fructose were equally effective calming agents, glucose slightly less so. Lactose, the milk sugar, was not at all effective and did not reduce crying any more than did water. In fact, some babies cried more when given lactose. A 2nd experiment established quantitative, dose-response functions for sucrose as a calming agent: 0.17M, 0.42M, and 0.51M sucrose reduced crying equally effectively. Moreover, crying reduction was not differentially affected by the volume of ingested sucrose, because 0.2 ml of 0.34M sucrose was as effective as 0.6 or 1.0 ml of 0.34M sucrose. Results suggest that sucrose calms in a stepwise manner with a flat suprathreshold function and that the calming basis of milk must be sought in components other than its sugar. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

The impact of daycare environments on children''s mood and behavior

The effect of dehulling, soaking and soaking/cooking on sucrose, raffinose and stachyose in mature dry seeds of nine varieties of cowpea (Vigna unguiculata) and one variety of tropical African yam bean (Sphenostylis stenocarpa) were investigated. The results showed a progressive decrease in sucrose, raffinose, and stachyose contents. Soaking for 12 hours and cooking for 30 min eliminated most of the sucrose, raffinose and stachyose. The sugar contents in whole raw cowpea were sucrose 0.73-4.58%, raffinose 0.71-6.86% and stachyose 2.38-3.87%, and for tropical African yam bean sucrose 4.08%, raffinose 1.08% and stachyose 4.14% while the seeds soaked for 12 hours and cooked for 30 min had for cowpea sucrose 0.03-0.81%, raffinose 0.04-0.20% and stachyose 0.12-0.72%, and tropical African yam bean sucrose 0.70%, raffinose 0.40% and stachyose 0.41%.     

Prevention of progressive myopia

Changes in sugars and polyols content of fruits and leaves of three cultivar of peach, from fruit set to over maturity, have been studied. Sucrose, glucose, fructose and sorbitol were found both in leaves and in fruits. In fruits, inositol and galactose were also found but only in traces. Redhaven fruits show the greatest total sugars content (sorbitol included) (866 mg/g.d.m.)followed in order by Favorita and Prodigiosa ones (771 and 694 mg/g.d.m.) For Redhaven and Favorita fruits, at harvest matuity, the sugar present in greatest amount was sucrose followed in order by glucose, fructose and sorbitol. On the contrary, in Prodigiosa fruits the order was sucrose, sorbitol, fructose and glucose. In the leaves of three cultivar, the compound present in the greatest amount was sorbitol followed by sucrose, glucose and fructose. The sucrose decrease in leaves during fruit growth could confirm that the disaccharide is the main sugar transported from the leaves to the fruits.     

Detection of counterfeit and relabeled infant formulas by high-pH anion-exchange chromatography-pulsed amperometric detection for the determination of sugar profiles

High pH anion-exchange separation with pulsed amperometric detection (HPAE-PAD) is used to characterize various milk-based, soy-based, and protein hydrolysate infant formulas based on carbohydrate profiles. Counterfeit and relabeled formulas are compared to authentics. Figures of merit are shown for glucose, fructose, lactose, sucrose, and maltose.     

Taste in acceptance of sugars by human infants.

Offered 96 male and 96 female 23-84 hr old infants water and a solution of glucose, fructose, lactose, or sucrose for 3-min periods. Volumes ingested were measured. Ss discriminated between water and a solution of sugar, demonstrating a distinct preference for the latter. The effectiveness of sugars in evoking ingestion varied with both the compound and the solution concentration. (15 ref.) (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Topology of allosteric regulation of lactose permease

Sugar transport by some permeases in Escherichia coli is allosterically regulated by the phosphorylation state of the intracellular regulatory protein, enzyme IIAglc of the phosphoenolpyruvate:sugar phosphotransferase system. A sensitive radiochemical assay for the interaction of enzyme IIAglc with membrane-associated lactose permease was used to characterize the binding reaction. The binding is stimulated by transportable substrates such as lactose, melibiose, and raffinose, but not by sugars that are not transported (maltose and sucrose). Treatment of lactose permease with N-ethylmaleimide, which blocks ligand binding and transport by alkylating Cys-148, also blocks enzyme IIAglc binding. Preincubation with the substrate analog beta-D-galactopyranosyl 1-thio-beta-D-galactopyranoside protects both lactose transport and enzyme IIAglc binding against inhibition by N-ethylmaleimide. A collection of lactose permease replacement mutants at Cys-148 showed, with the exception of C148V, a good correlation of relative transport activity and enzyme IIAglc binding. The nature of the interaction of enzyme IIAglc with the cytoplasmic face of lactose permease was explored. The N- and C-termini, as well as five hydrophilic loops in the permease, are exposed on the cytoplasmic surface of the membrane and it has been proposed that the central cytoplasmic loop of lactose permease is the major determinant for interaction with enzyme IIAglc. Lactose permease mutants with polyhistidine insertions in cytoplasmic loops IV/V and VI/VII and periplasmic loop VII/VIII retain transport activity and therefore substrate binding, but do not bind enzyme IIAglc, indicating that these regions of lactose permease may be involved in recognition of enzyme IIAglc. Taken together, these results suggest that interaction of lactose permease with substrate promotes a conformational change that brings several cytoplasmic loops into an arrangement optimal for interaction with the regulatory protein, enzyme IIAglc. A topological map of the proposed interaction is presented.     

Addition of an osmotic agent to liver preservative solutions in a model of in vitro preservation of hepatocytes

To avoid hypoxic cell swelling during liver preservation followed by reduced perfusion in the reoxygenation period, osmotic substances such as mannitol, sucrose and raffinose, and the impermeant anion lactobionate are used in established liver preservation solutions. The various osmotic agents were investigated at concentrations of 60, 140, 260 and 300 mM, the solutions being kept isotonic by substitution with sodium and potassium chloride to 300 mosmol/l. Cultures of adherent pig hepatocytes were incubated in an in vitro model of cold hypoxia (4 degrees C, PO2 < 0.1 mmHg) for 24 h and reoxygenated with standard culture medium for 3 h. After each incubation period, light microscopy was performed to estimate cell viability and detachment rate. LDH and GOT liberation were also measured. To estimate the change in cell volume, isolated hepatocytes were incubated in suspension for 24 h of cold hypoxia. The cell volumes were compared after centrifugation and measurement of the pellet and the solute levels. Rising concentrations of osmotic substances resulted in increasing liberation of LDH and GOT. The levels of LDH and GOT release from cultures incubated with 60 mmol/l sucrose or raffinose were comparable to those in a preservation solution of "extracellular" ion composition. Addition of mannitol to the preservation solution resulted in cell damage. At high concentrations, sucrose did not affect the hepatocytes as much as raffinose. While mannitol can permeate the hepatocytes and lead to cell swelling, a cell-shrinking effect was observed when sucrose was used, and even more pronounced cell shrinking was seen with raffinose, to which the hepatocyte membrane is known to be permeable.(ABSTRACT TRUNCATED AT 250 WORDS)     

Preliminary investigations of an amperometric oligosaccharide dehydrogenase-based electrode for the detection of glucose and some other low molecular weight saccharides

Biosensors for the determination of sugars were constructed using oligosaccharide dehydrogenase (ODH) and soluble phenazine methosulfate (PMS) or an osmium-based three-dimensional redox hydrogel. In the latter case the enzyme and poly(1-vinylimidazole) complexed with osmium (4,4'-dimethylbpy)2Cl were cross-linked with poly(ethylene glycol) diglycidyl ether. Both electrode configurations showed similar sensitivities for glucose in the range between 8 and 21 muAmM-1 cm-2. The responses for 10 mono and oligosaccharides were studied. There was no response for fructose. In the concentration range 0.1-20 mM the relative sensitivities were determined for arabinose (96%), xylose (3%), mannose (50%), galactose (11%), glucose (100%), maltose (24%), lactose (12%), cellobiose (34%) and maltotriose (10%).     

Disturbed galactose metabolism in elderly and diabetic humans is associated with cataract formation

Lactose consumption has been associated with a high incidence of cataract in northern Indian and southern Italian populations. Galactose absorbed after hydrolysis of lactose from milk in individuals with normal lactase activity is considered responsible. However, lactase-deficient subjects who often avoid drinking milk are able to digest lactose and absorb free galactose in fermented milk and yogurt. This study was conducted to evaluate the relationships between milk and yogurt consumption, galactose metabolism and cataract risk. Milk ingestion was dose-related with cataract risk in lactose digesters (particularly in diabetics) but not in lactose maldigesters. Conversely, yogurt intake had a protective dose-effect on cataract formation for the whole population. Maximal galactose concentrations after an oral galactose test increased exponentially with age. Red blood cell galactokinase activity was significantly lower in elderly subjects (> 60 y) than in young individuals (P < 0.05), and galactose-1-phosphate uridyl-transferase activity was significantly lower in institutionalized subjects and in home-living elderly with cataract than in healthy elderly subjects (P < 0.05). We conclude that the cataractogenic action of milk lactose is dependent on the disturbance of galactose metabolism in elderly subjects and that yogurt is not cataractogenic, although the mechanism of the protective effect of yogurt remains unknown.     

Sucrose and fructose have qualitatively different flavors to rats

Previous studies suggest that rats might be able to discriminate between sucrose and fructose, but no previous study has examined this possibility in much detail. Rats were conditioned to avoid either sucrose or fructose by injecting them with lithium chloride when they drank these substances. Control rats were given the same injections but were not exposed to either sugar during training. After training, the rats were given a choice of fructose vs. sucrose. Data from control rats provided information about the relative taste intensity of the sugars. If the sugars possess only a single gustatory quality, control rats should prefer the sweeter sugar; under this assumption, sucrose appears to be two-four times sweeter than fructose. The two sugars share a common taste because rats trained to avoid sucrose avoided fructose when the fructose concentration was much greater than the sucrose concentration. Nevertheless, the two sugars are discriminable because, when the apparent sweetness of the sugars was matched, rats showed a greater aversion to the sugar they were trained to avoid. Aversions to sucrose and fructose also generalized to maltodextrins, but sucrose may have a somewhat greater maltodextrin flavor than does fructose. It is proposed that the biological function of maltodextrin taste is to allow animals to sense the ratio of glucose to fructose in foods.     

Kinetics of plasma fructose and glucose when lactose and fructose are used as energy supplements for neonatal calves

Shortly after birth, plasma glucose and fructose concentrations of the neonate decline and thus leave blood sugar below the homeostatic mode. Two trials were conducted to determine the plasma glucose and fructose kinetics in control and supplemented calves for 108 h after birth. In the short-term trial, six Holstein calves were given 40 g of either fructose, lactose, or water (control) orally at 1 and 96 h after birth. Treatments were administered with a colostrum substitute (Life Boost) at 1 h and whole milk at 96 h. Rectal temperatures and changes in plasma glucose and fructose concentrations were monitored at close intervals for 12 h after supplementation. In the long-term trial, 15 Holstein calves were given 40 g of either lactose, fructose, or water (control) at 1 h after birth and at 12-h intervals for 81 h. Plasma glucose and fructose concentrations were determined before and 4 h after each of the seven feedings. Early postpartal feeding of fructose suppressed plasma glucose (approximately 50%), with a reciprocal rise in plasma fructose. Irrespective of treatment, plasma glucose concentrations did not stabilize (approximately 100 mg/dL) until 17 to 24 h after birth. After 24 h, lactose supplements increased concentrations of plasma glucose 4 h after supplementation (169.7 +/- 8.2 mg/dL), compared with those in calves that did not receive the additional lactose. After 24 h, fructose supplements did not affect plasma glucose, but plasma fructose concentrations increased (82.6 +/- 12.4 mg/dL) 4 h after administration. The response to fructose supplements declined by 11.4 mg x dL(-1) x d(-1). Fructose was not detected in the plasma of control or lactose-treated calves after 17 h after birth. Calves that received fructose supplements had rectal temperatures 8 and 10 h after birth that were higher than those of the other calves. The mechanisms of sugar metabolism change quickly following birth. Oral sugar supplements increase the total plasma sugar concentrations of treated calves.     

Identification of a new porin, RafY, encoded by raffinose plasmid pRSD2 of Escherichia coli

The conjugative plasmid pRSD2 carries a raf operon that encodes a peripheral raffinose metabolic pathway in enterobacteria. In addition to the previously known raf genes, we identified another gene, rafY, which in Escherichia coli codes for an outer membrane protein (molecular mass, 53 kDa) similar in function to the known glycoporins LamB (maltoporin) and ScrY (sucrose porin). Sequence comparisons with LamB and ScrY revealed no significant similarities; however, both lamB and scrY mutants are functionally complemented by RafY. Expressed from the tac promoter, RafY significantly increases the uptake rates for maltose, sucrose, and raffinose at low substrate concentrations; in particular it shifts the apparent K(m) for raffinose transport from 2 mM to 130 microM. Moreover, RafY permits diffusion of the tetrasaccharide stachyose and of maltodextrins up to maltoheptaose through the outer membrane of E. coli. A comparison of all three glycoporins in regard to their substrate selectivity revealed that both ScrY and RafY have a broad substrate range which includes alpha-galactosides while LamB seems to be restricted to malto-oligosaccharides. It supports growth only on maltodextrins but not, like the others, on raffinose and stachyose.