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Acquisition of ability to utilize Xylitol: disadvantages of a constitutive catabolic pathway in Escherichia coli.

Ribitol+ strains of Escherichia coli acquire the ability to utilize xylitol by mutating to constitutive production of the coordinately controlled ribitol catabolic enzymes ribitol dehydrogenase (RDH) and D-ribulokinase (DRK). Such strains concomitantly acquire toxicity to galacitol and L-arabitol, and to D-arabitol if they are unable to utilize it for growth. Strains selected for resistance to these polyols have DRK structural gene mutations or other mutations that eliminate the constitutive production of DRK, consistent with the view that DRK phosphorylates those polyols to toxic substances. Ribitol+ strains selected for growth on 8 mM xylitol fail to grow on 30 mM xylitol. A product of ribitol and xylitol catabolism represses synthesis of RDH, an enzyme required for growth on xylitol. At 30 mM xylitol, greater than 99% of RDH synthesis is repressed. Strains that grow on 8 mM xylitol can mutate to grow on 30 mM xylitol. Such mutants, relieved of this repression, overproduce RDH, resulting in good growth on the poor substrate, xylitol, but poor growth on the normal substrate, ribitol.

Escherichia coli

Shuttles for translocation of NADH in isolated liver cells from fed rats during oxidation of xylitol.

Translocation of xylitol-derived NADH via malate-aspartate and alpha-glycerophosphate shuttles was studied in liver cells isolated from fed rats. In bicarbonate medium amino-oxyacetate, rotenone and antimycin A, were equally efficient in depressing the xylitol removal. Incubation of cells in nonbicarbonate medium did not affect the rate of xylitol removal. In this medium amino-oxyacetate and antimycin A, but not rotenone, inhibited xylitol removal. Xylitol inhibited the lactate accumulation found when the cells were incubated without any exogenous substrates. Glucose was the main end product of xylitol oxidation. In nonbicarbonate medium ketogenesis was high, whereas in bicarbonate medium a low rate of ketone body formation was found. Xylitol had no effect on the rate of ketone body formation in either medium tested. Xylitol markedly decreased the ATP and Pi contents of the cells, but no change in the ATP/ADP x Pi ratio or the rate of oxygen consumption was found. The results suggest that NADH formed during xylitol oxidation is translocated to the mitochondria mainly through the malate-aspartate shuttle and only when this shuttle is inhibited does the alpha-glycerophosphate shuttle transfer NADH. Intramitochondrial reactions which form NADH and FADH2 are also suggested to be important regulators of the activity of the alpha-glycerophosphate shuttle.

Animals

Xylitol and D-arabitol toxicities due to derepressed fructose, galactitol, and sorbitol phosphotransferases of Escherichia coli.

d-Arabitol was observed to be toxic to many laboratory strains of Escherichia coli K-12, and xylitol was found to be toxic to an existing E. coli C mutant strain. Fructose-specific components of the phosphoenolpyruvate:sugar phosphotransferase system are required for xylitol toxicity. Selection for xylitol resistance results in Fru(-) strains blocked in fructose phosphotransferase. Introduction of the ptsF or ptsI mutation into a xylitol-sensitive strain eliminates sensitivity. [(14)C]fructose uptake experiments imply that the mutation to xylitol sensitivity, which is co-transducible with ara and leu, results in derepression of normally inducible fructose phosphotransferase. Wild-type strains also become xylitol sensitive if induced by (and then removed from) fructose. Xylitol toxicity is prevented by fructose in both wild-type and mutant strains. Circumstances causing xylitol, a new food additive, to become toxic to an otherwise insensitive wild-type organism have not been reported previously. The d-arabitol-sensitive laboratory strains are galactitol (dulcitol) utilizers, although most other strains are not. Selection for d-arabitol resistance results in Gat(-) strains blocked in a constitutive galactitol-specific component of the phosphotransferase system. A mutation causing d-arabitol sensitivity occurred many years ago in AB284, the parent of AB311, AB312, AB313, and many other strains. d-Arabitol sensitivity also occurs in sorbitol-constitutive strains and is shown, like the previous two instances of pentitol toxicities, to result from a constitutive phosphotransferase, which is blocked in mutants selected for resistance.

Arabinose

Plaque growth while chewing sorbitol and xylitol simultaneously with sucrose flavored gum.

In a recent study, sorbitol flavored chewing gum was found neither to increase nor decrease the normal rate of plaque formation, whereas high plaque scores were obtained with sucrose gum during 4 days of no mechanical tooth cleaning. The aim of the present study was to see if chewing sorbitol or xylitol flavored gum together with sucrose gum would affect the growth rate of plaque and whether chewing of xylitol flavored gum could reduce the amount of already formed plaque. Twenty-seven dental students refrained from mechanical oral hygiene measures from Monday to Friday morning for 3 weeks. The students were randomly divided into three groups. A three time crossed-over double-blind approach was used. During each test period one group chewed a combination of one piece sorbitol and one piece sucrose flavored gum five times per day, the second group correspondingly chewed xylitol and sucrose flavored gum, while the third group served as a no hygiene control group. After each test period the students in the control group chewed one piece of xylitol gum every 15 minutes for 2.5 hours. The participants started out each week with clean teeth and were at the end of each test period scored for visible plaque on the facial, mesial and lingual surfaces of their teeth. There was somewhat more plaque after 4 days of chewing sucrose-sorbitol and sucrose-xylitol gum combinations than after no oral hygiene alone. There was no difference between the two test treatments. The 2.5-hour chewing of xylitol flavored gum after the no oral hygiene period did not result in a reduction of the 4-day-old plaque.

Chewing Gum

Effect of peroral administration of xylitol on exocrine secretions of monkeys.

The effects of the administration of xylitol on the biochemical properties of monkey parotid and submandibular saliva and lacrimal fluid were investigated. Monkeys (Macaca mulatta) were fed either a sucrose or xylitol diet for 3 days. Ingestion of xylitol was associated with a significant increase of the activity of the salivary lactoperoxidase. The concentration of protein was also increased. Higher lactoperoxidase activity was found in parotid than in submandibular saliva. The concentrations of inorganic phosphate, calcium and SCN ions were practically unaffected. The concentration of SCN ions in pilocarpine stimulated monkey saliva was low (less than 5.5 mg/liter). Peroral administration of 2.5 g of xylitol or sorbitol per day to M. fascicularis resulted in almost similar levels of salivary lactoperoxidase activity. The administration of xylitol orally or by gastric intubation was not found to affect the concentration of lactoperoxidase, protein, phosphate, and SCN and iodine ions in lacrimal fluid. The results suggest that specific dietary sugars have a selective effect on the biochemical properties of saliva.

Animals

Effect of 4.5--year use of xylitol and sorbitol on plaque.

The pH values of xylitol-containing plaque suspensions of persons who habitually used xylitol and sorbitol during 3.2 to 4.5 years did not significantly differ from those observed with suspensions containing no added carbohydrates. Sorbitol produced in 22-hour incubations pH values as low as 3.9 to 5.5. The activity of plaque xylitol dehydrogenase was almost nil, whereas sorbitol dehydrogenase activity was higher. The nonacidogenic nature of xylitol in relation to plaque did not change in prolonged use of xylitol.

Adolescent

[The cariogenicity of xylitol in the animal experiment].

After programmed feeding of rats in a six and eight-week long conventional experiment with increasing concentrations of xylitol, compared to sorbitol, fructose and saccharose, the non-cariogenic nature of xylitol was confirmed. The increasing amounts of xylitol after sorbitol in chocolate diets (up to 30 g/day/rat) led to serious dilatation of the cecum and to changes in the mucosa of cecum and colon when sorbitol was given. Examination of plaques of the germ-free rats monoassociated with S. mutans showed that xylitol had no bacteriostatic effect on this type of germ. Since xylitol is not broken down by these germs with acid being formed, careis did not continue to extend.

Animals

[Further improvement of the ACD-AG protective solution for blood. III. Improvement of the oxygen transport function of erythrocytes in sorbitol xylitol pyruvate solutions with elevated Ph].

If the pH value in the ACD or in the ACD-AG storage solution is enhanced, the glucose in the autoclaving with undergo a caramelizing process. For this reason glucose was replaced by sorbite in the storage solutions with a pH value of 6.0 and additions of xylitol and pyruvate. The initial pH value in the blood amounted to 7.3. The content of 2.3 DPG of the erythrocytes remained fully preserved in the blood with sorbitol and additions of xylitol and pyruvate during the first 2 weeks of storage and decreased to 30% only in the third week. There were only slight amounts of 2.3 DPG in the ACD-AG blood at that time of storage. Up to the third week of storage the ATP content of erythrocytes as well as the haemoglobin level in the plasma revealed no essential differences between stored blood with sorbitol and xylitol as a substrate or glucose + xylitol respectively. The quick decrease of the ATP level to zero and the simultaneous strong increase of haemolysis in the sorbitol blood within the fourth week of storage is discussed in connection with a lowering of the NAD/NADH2 quotient. For the purpose of keeping the 2.3 DPG level of erythrocytes a storage solution with sorbite and xylitol (ASCX-AG-Pyr 10mM) seems to be well suited for a storing time of 2---3 weeks at first.

Adenosine Triphosphate

Growth of a mutant of Escherichia coli K-12 on xylitol by recruiting enzymes for D-xylose and L1,2-propanediol metabolism.

Wild type Escherichia coli K-12 cannot grow on xylitol and we have been unsuccessful in isolating a mutant directly which had acquired this new growth ability. However, a mutant had been selected previously for growth on L-1,2-propanediol as the sole source of carbon and energy. This mutant constitutively synthesized a propanediol dehydrogenase. Recently, we have found that this dehydrogenase fortuitously converted xylitol to D-xylose which could normally be metabolized by E. coli K-12. In addition, it was also discovered that the D-xylose permease fortuitously transported xylitol into the cell. A second mutant was thus isolated from the L-1,2-propanediol-growing mutant that was constitutive for the enzymes of the D-xylose pathway. This mutant could indeed grow on xylitol as the sole source of carbon and energy, by utilizing the enzymes normally involved in D-xylose and L-1,2-propanediol metabolism.

Alcohol Oxidoreductases

Metabolic investigations after xylitol infusion in human subjects.

Evidence has been sought for minor degrees of thiamin and pyridoxine deficiency in patients undergoing surgery who have been infused with xylitol as a parenteral nutrient. Some metabolic changes which are associated with this practice have been studied; the findings are compared with those obtained in similar patients infused with glucose solutions. The thiamin status of all of the subjects was normal. Some of the patients showed slight biochemical evidence of pyridoxine deficiency, but there were no untoward effects of xylitol infusion. The concentration of oxalate in the blood and the excretion of oxalate in the urine did not exceed the normal range in any patient. The plasma and urine orthophosphate and urinary pyrophosphate levels decreased in association with the infusion of both xylitol and glucose. Plasma pyrophosphate and calcium levels, and the urinary calcium level, were essentially unaltered. A detailed quantitative study of the urinary organic acid excretion by means of gas chromatography/mass spectrometry showed that there was an abnormal glycolic aciduria and tetronic aciduria associated with xylitol infusion, but not with glucose infusion. There was no evidence of increased oxalate excretion in any patient by this method. The biochemical and clinical significance of these findings is discussed.

Adult

Effect on plaque growth of xylitol and sucrose-containing chewing gums.

Two experiments tested the effects of xylitol or sucrose-containing chewing gums on plaque formation. In the first experiment the 18 subjects maintained their normal dietary habits, and in the second they received a sucrose-free diet during the 3-day experimental periods. Plaque formation was assessed gravimetrically and/or planimetrically. When the dietary habits were not altered, neither the sucrose nor the xylitol-containing chewing gums had significant effects on the plaque scores. When sucrose was almost eliminated from the diet, plaque formation was significantly higher in the sucrose than in the xylitol and the placebo groups. Although plaque formation was lower in the xylitol group than in the placebo group, the differences were not statistically significant.

Adult

Turku sugar studies XX. Microbiological findings and plaque index values in relation to 1-year use of xylitol chewing gum.

The aim was to study possible alterations in the microbial flora of plaque and saliva in relation to partial substitution of dietary sucrose with xylitol. The development of plaque index values was observed simultaneously. These observations were carried out during a 1-year clinical trial, the effects of sucrose (S) and xylitol (X) chewing gum on the incidence of dental caries being observed in 100 young adults. Paraffin-stimulated saliva samples were diluted stepwise and cultivated on Rogosa S.L. agar and Sabouraud agar aerobically. Lyophilized dental plaque samples were cultivated on phenol red agar under anaerobic and aerobic conditions. The pH-values were measured after incubating the mixed plaque flora for 1 and 7 days in the presence of various sugars. Both the arithmetic and geometric means of the total CFU values on Rogosa S.L. agar decreased in the S-group at the 6-month phase but returned to the starting level after one year, whereas in the X-group they decreased or remained on the starting level. At the 6-month phase the difference between the groups was significant (U-test, p = 0.0013) and almost significant (U-test, p = 0.0569) at the end of the study. No significant differences or changes could be seen between or within the groups on Sabouraud agar. The geometric mean values of S. sanguis and S. mutans as well as the total CFU values on phenol red agar decreased considerably in both the S- and X-groups, but no significant differences could be detected in any of the streptococcal counts between the groups. The pH of the carbohydrate-containing culture media infected with mixed dental plaque significantly decreased, with the exception of the xylitol containing ones in which the pH values were not lowered even after 7 days' incubation. A significant decrease in plaque formation in relation of chewing per se was demonstrable. The difference in the plaque index values equalling or exceeding 2 was significant between the S- and X-groups. No bacterial adaptation to utilize xylitol occurred during the trial.

Chewing Gum

Effects of 3 months frequent consumption of hydrogenated starch hydrolysate (Lycasin), maltitol, sorbitol and xylitol on human dental plaque.

Lozenges containing hydrogenated starch hydrolysate (Lycasin), maltitol, sorbitol or xylitol were consumed 4 times daily during 3 months by 4 groups of persons (in all 85 subjects). In the maltitol-, sorbitol- and xylitol-group the plque wet weights were of the same magnitude before and after the test period. In the Lycasin-group, a higher value was found after than before the 3-month period (p less than 0.01). The acid production in suspensions of dental plaque material from Lycasin, maltitol and sorbitol expressed as per cent of that from glucose was approximately the same before and after the test period. From xylitol no acid production could be demonstrated either before or after the 3-month period. There were no statistically significant differences between the plaque pH-changes induced by rinsing with 50% solutions of Lycasin, maltitol, sorbitol or xylitol before and after the test period. However, there was a tendency (p less than 0.05) towards lower pH-values induced by the maltitrol and sorbitol rinse after the 3-month period compared with before. No difference in the relative numbers of facultative anaerobic streptococci. Streptococcus mutans or facultative anaerobic lactobacilli before and after the test period was found.

Adult

[Contribution to the microbial catabolism of xylitol].

Five xylitol metabolizing streptococci strains from the oral cavity of the rat were examined with regard to their metabolic capacities for xylitol, sorbitol, mannitol, fructose, glucose and saccharose by means of the Warburg technique. The amounts metabolized were smallest with xylitol and sorbitol and considerably bigger with mannitol and fructose. By far the highest CO2 values were obtained with glucose, while saccharose was not metabolized at all. The experimental results confirm the weak metabolic capability of these organisms with regard to the sugar substitute xylitol.

Animals

Dissolution rate of p-aminobenzoates from solid xylitol dispersions.

Xylitol was studied as a carrier in solid dispersions because of its low melting point and stability up to 180 degrees. It is more stable than sucrose and does not enter into Maillard reactions. Solid dispersions were prepared from esters of p-aminobenzoic acid and xylitol by the melting method and were compressed into tablets. The p-aminobenzoate dissolution rates were determined by a modified beaker method. The increase in the dissolution rates was greatest at the lowest drug levels. When the dispersion drug content exceeded 20-30%, the dissolution rate per unit area remained nearly constant. In the latter case, the increase in the dissolution rate was primarily due to an increase in area. When the carbon chain length was increased in the homologous series, the dissolution rate from the xylitol dispersions showed a nearly linear decrease.

4-Aminobenzoic Acid

Xylitol associated changes in amylase and protein content of monkey parotid saliva.

The purpose of this study was to examine amylase activity in the parotid saliva of monkeys fed a high sugar diet supplemented for 3-day periods with either sucrose or xylitol. All monkeys fed xylitol displayed a significant elevation of amylase activity and total protein content in their parotid saliva. The specific activity of amylase (units/mg protein) did not differ between the two diets. The possible importance of this observation to the purported cariostatic action of xylitol is discussed.

Amylases

Turku sugar studies XXI. Xylitol, sorbitol-, fructose- and sucrose-induced physico-chemical changes in saliva.

The aim was to study eventual physico-chemical changes occurring in whole saliva due to sweetened and unsweetened stimulators. The assay was carried out in 10 female subjects with regard to changes of pH, buffering capacity and electrolytes in saliva as influenced by chewing of fructose, sucrose, sorbitol and xylitol gum, gum base and paraffin. The flow rate of saliva was measured in relation to use of xylitol and sucrose chewing gum and unsweetened gum base. These sweeteners increased significantly the salivary flow rate in comparison to the unsweetened gum base. Generally, xylitol and sorbitol on one hand, and sucrose and fructose on the other, behaved in an almost similar way. Increased buffering capacity and elevation of pH saliva was found in the presence of the polyols tested.

Bicarbonates

[Use of maltose and a mixture of maltose, fructose and xylitol in parenteral feeding].

Maltose or maltose in combination with fructose and xylitol was administered intravenously to eight healthy male subjects. Constant maltose levels could not be attained in the blood at an infusion rate of 0.125 g maltose/kg body-weight and hour. Maximal concentrations of maltose were found at the end of the infusion period. 8.6 +/- 1.2% of the administered radioactivity was excreted into urine within 8 hours. Regarding the enzymatically determined maltose and glucose, the maltose balance was more favorable with the loss of only 3--3.5% carbohydrates into urine. The highest oxidation rate of the administered maltose was 1.5 g maltose/human volunteer and hour. During the experimental period of 8 hours 7.4 g maltose, corresponding to 37% of the applied dosis of the disaccharid, has been oxidized to and excreted as 14CO2. Xylitol and fructose did not effect utilisation and balance of maltose. Only the urinary excretion of glucose was higher when the combined solution was applied. At a limited infusion rate (0.125 g maltose/kg body-weight and hour) maltose or the combined solution maltose--xylitol--fructose may be recommended for parenteral nutrition.

Adult