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PubMed2004

Identification of sorbitol transporters expressed in the phloem of apple source leaves.

Sorbitol is a major photosynthetic product and a major phloem-translocated component in Rosaceae (e.g. apple, pear, peach, and cherry). We isolated the three cDNAs, MdSOT3, MdSOT4, and MdSOT5 from apple (Malus domestica) source leaves, which are homologous to plant polyol transporters. Yeasts transformed with the MdSOTs took up sorbitol significantly. MdSOT3- and MdSOT5-dependent sorbitol uptake was strongly inhibited by xylitol and myo-inositol, but not or only weakly by mannitol and dulcitol. Apparent K(m) values of MdSOT3 and MdSOT5 for sorbitol were estimated to be 0.71 mM and 3.2 mM, respectively. The protonophore, carbonyl cyanide m-chlorophenylhydrazone (CCCP), strongly inhibited the sorbitol transport. MdSOT3 was expressed specifically in source leaves, whereas MdSOT4 and MdSOT5 were expressed in source leaves and also in some sink organs. MdSOT4 and MdSOT5 expressions were highest in flowers. Fruits showed no or only weak MdSOT expression. Although MdSOT4 and MdSOT5 were also expressed in immature leaves, MdSOT expressions increased with leaf maturation. In addition, in situ hybridization revealed that all MdSOTs were expressed to high levels in phloem of minor veins in source leaves. These results suggest that these MdSOTs are involved in sorbitol loading in Rosaceae.

Amino Acid Sequence↗
PubMed1987

Sorbitol intolerance in adults. Prevalence and pathogenesis on two continents.

The prevalence and pathogenesis of sorbitol intolerance in adults have not been adequately studied. On oral administration of sorbitol (10 g), 32% of the 124 healthy adults (41 in the U.S.A., 83 in India) developed abdominal symptoms. The orocecal transit time, measured in Asian Indian volunteers by breath hydrogen (H2) analysis, was significantly shorter in those intolerant to sorbitol (71.8 +/- 36.9 min) than in those who were tolerant (109.5 +/- 47.8 min). Our results indicate that sorbitol intolerance is a potential clinical problem in a substantial number of healthy adults. A short orocecal transit time may be responsible for sorbitol intolerance in some people. We discuss the sorbitol content of common foodstuffs.

Adult↗
PubMed1993

Apple juice malabsorption: fructose or sorbitol?

Carbohydrate malabsorption after apple juice ingestion may produce abdominal symptoms and diarrhea, especially in children. The carbohydrates suggested to play roles in this process are fructose, as it is present in excess of glucose, and sorbitol. Absorption of the carbohydrates in apple juice was investigated in 17 children and 12 adults by means of the hydrogen breath test. Apple juice was given at a dose of 15 ml/kg body weight, with a maximum of 375 ml. Fructose (0.6 g/kg) and sorbitol (0.06 g/kg), alone and in combination, were administered in amounts similar to their contents in apple juice (fructose as excess over glucose content). Apple juice malabsorption, as judged by a peak breath H2 excretion of > or = 20 ppm, was found in 11 children (65%) and 4 adults (33%). Of those malabsorbing apple juice, 7 of 11 children malabsorbed fructose, 1 of 11 sorbitol, and 4 of 11 the combination; the four adults absorbed all test solutions completely. We could not find an additive effect of sorbitol on breath H2 excretion after fructose ingestion. Peak breath H2 concentrations after apple juice ingestion (mean +/- SEM: 43 +/- 7 ppm) were higher than those with fructose (23 +/- 5 ppm; p < 0.05) or the fructose-sorbitol combination (20 +/- 5 ppm; p < 0.05). Fructose, and not sorbitol, is the sugar responsible for the increase in breath H2 after apple juice consumption and therefore for the diarrhea accompanying excessive apple juice consumption in toddlers.

Adult↗
PubMed1996

Facilitating effect of amino acids on fructose and sorbitol absorption in children.

The facilitating effect of glucose on free fructose absorption has been suggested to be due to a sucrase-related transport mechanism. In contrast, the conditions influencing the absorption of sorbitol have hardly been investigated. As amino acids promote transcellular water flow, we investigated their effects on the absorption of fructose and sorbitol. We studied 15 healthy children using breath hydrogen tests following the ingestion of fructose and sorbitol, alone and in combination with glucose or amino acids. Similarly, the effect of acarbose pretreatment on sucrose and fructose-glucose absorption was investigated. The inhibition of sucrase isomaltase by acarbose impedes the absorption of sucrose but not of the fructose-glucose mixture. Fructose absorption is enhanced by glucose and by the amino acids L-alanine, L-glutamine, L-phenylalanine, and L-proline. Similarly, the absorption of sorbitol is facilitated by glucose and L-alanine. These results are not in concordance with a sucrase-related fructose-transport system and suggest another mechanism for glucose-induced enhancement of fructose (and sorbitol) absorption. We hypothesize that the absorption of fructose and sorbitol may be stimulated by the increased water flux induced by active absorption of glucose as well as amino acids.

Adolescent↗
PubMed1996

A simple and rapid method for the determination of D-sorbitol in plasma using the Cobas Mira S.

The standard enzymatic assay for quantification of D-sorbitol in plasma was adapted to the automatic analyzer Cobas Mira S. In the assay, NAD (reagent) in the presence of sorbitoldehydrogenase (SDH; start reagent) converts D-sorbitol to fructose with formation of NADH, which was detected automatically as the difference between the first and last readings at 340 nm. The sample blank values for each specimen were subtracted to exclude both endogenous D-sorbitol and sugars, which also react as substrates for SDH. The method is simple, rapid (40 samples/h), precise down to endogenous concentrations (coefficient of variation < 5%; limit of determination: 0.38 mg/L) and linear up to 100 mg/L. Samples with higher D-sorbitol concentrations were estimated after dilution. The method was used to measure disposition curves of sorbitol in volunteers after a single intravenous dose of 0.8 g sorbitol.

Humans↗
PubMed1992

Purification and properties of sorbitol-6-phosphate dehydrogenase from oral streptococci.

The activity of sorbitol-6-phosphate (S6P) dehydrogenase (S6PDH) and the sorbitol transport system were studied in strains of the oral streptococci Streptococcus gordonii, Streptococcus mitis, Streptococcus sanguis and Streptococcus mutans. Genetically transformed (to ferment sorbitol) strains and their DNA donors were included. S6PDH was purified by anion exchange chromatography and gel filtration. The purity of the enzyme was confirmed by polyacrylamide gel electrophoresis. The purified enzyme from all the strains exhibited Michaelis-Menton saturation kinetics. The Km values for nicotinamide-adenine dinucleotide (NAD) and S6P ranged between 0.03 and 0.21 mM and 0.07 and 0.20 mM respectively. The relative molecular weights of the native enzyme were 229,000 for one donor-transformant pair (S. sanguis and S. gordonii), 107,000 for the other pair (S. mitis and S. gordonii) and 129,000 for S. mutans. The molecular weights of the S6PDH subunits ranged from 26,000 to 28,000. The pH optima (greater than 8.5) and the amino acid composition (15 amino acids examined) were similar for the S6PDH from the different strains. However, the chromatographic and electrophoretic patterns as well as the Km values for NAD and S6P were the same only between the S6PDHs from the strains within each donor-transformant pair. Purified S6PDH from S. mutans also exhibited low mannitol-1-phosphate dehydrogenase activity. Sorbitol-grown decryptified cells of all the strains phosphorylated sorbitol in the presence of phosphoenolpyruvate but not in the presence of adenosine triphosphate (ATP). ATP-mediated phosphorylation of glucose was observed with the same strains when grown on glucose. No evidence for a non-phosphotransferase transport system was found for sorbitol in any of the strains.(ABSTRACT TRUNCATED AT 250 WORDS)

Biological Transport, Active↗
PubMed1994

The role of the succinate pathway in sorbitol fermentation by oral Actinomyces viscosus and Actinomyces naeslundii.

The sorbitol fermentation by Actinomyces viscosus and Actinomyces naeslundii was studied with washed sorbitol-grown cells. The fermentation was followed by titration of acids produced at pH 7.0 under anaerobic conditions. Metabolic end-products and intracellular levels of NAD, NADH and glycolytic intermediates during the fermentation were also analyzed. Cell extracts were examined for certain enzyme activities. Bicarbonate was required for acid production from sorbitol and from a mixture of glucose and sorbitol. Malate and fumarate could also support the acid production of A. viscosus. The main end-products were succinate and lactate but not ethanol. Cell extracts showed no activities of alcohol and aldehyde dehydrogenases, but they had activities of malate dehydrogenase and fumarate reductase. In the absence of bicarbonate, malate or fumarate, the intracellular NADH/NAD ratio increased and the levels of 3- and 2-phosphoglycerate and phosphoenolpyruvate decreased. The results indicate that oral sorbitol-fermenting actinomyces lack the ethanol pathway that can contribute to NADH oxidation. To maintain intracellular redox balance during anaerobic sorbitol fermentation, these bacteria can oxidize surplus NADH through a succinate pathway.

Actinomyces↗
PubMed1995

Effect of acetate on sorbitol fermentation by oral lactobacilli.

The rate of acid production and end-products from sorbitol were measured under anaerobic conditions in washed-cell suspensions of oral strains of Lactobacillus casei subsp. casei and Lactobacillus casei subsp. rhamnosus. The enzymatic activities were assayed in cell extracts of these strains. The cells fermented sorbitol to lactate, formate, ethanol and acetate under anaerobic conditions. Exposure of the cells to air (oxygen) led to inactivation of pyruvate formate-lyase and inhibition of anaerobic sorbitol fermentation. In the presence of acetate, air-exposed cells fermented sorbitol with a concomitant consumption of acetate and production of ethanol and lactate. Acetate also enhanced acid production from sorbitol in cells kept under anaerobic conditions and resulted in formation of lactate and ethanol. Cell extracts of all the strains had NADH-coupled acetate-reducing activity, which consisted of sequential reactions of acetate kinase, phosphotransacetylase, acylating aldehyde dehydrogenase and alcohol dehydrogenase. These findings indicate that oral lactobacilli can utilize acetate as an electron acceptor for maintaining their intracellular redox balance during anaerobic sorbitol fermentation in the absence of pyruvate formate-lyase activity.

Acetate Kinase↗
PubMedDate not supplied

Red cell sorbitol levels during hyper and hypoglycaemic clamp studies in insulin-dependent diabetic patients.

The red cell sorbitol concentration has been suggested as a measure of polyol pathway activity. Red cell sorbitol levels were higher in 53 patients having insulin-dependent diabetes mellitus (IDDM) than in 16 control subjects. Six patients having IDDM underwent hyperglycaemic 'clamp' studies; the red cell sorbitol level returned to the normal range when the blood glucose was clamped at 5 mmol/l for 1 h and rapidly increased when it was clamped at 15 and 25 mmol/l for a further hour at each level. Seven patients with IDDM were rendered hypoglycaemic; red cell sorbitol levels rapidly fell to a level less than, but not significantly different from normal. The results of these studies suggest that in IDDM red cell sorbitol levels are a reflection of prevailing blood glucose concentration and do not indicate long-term sorbitol accumulation in other tissues.

Adult↗
PubMed1989

Solid state adsorption of antibiotics onto sorbitol.

The ability of two types of sorbitol, instant and crystalline, to hold antibiotics permanently after mixing has been assessed by an air sieving technique. Sorbitol instant was found to have a greater adsorption capacity and binding strength than crystalline sorbitol. The six antibiotics studied were found to fall roughly into two groups of different adsorption capacities: (1) pivampicillin, cephalexin monohydrate and erythromycin ethylsuccinate, and (2) ampicillin trihydrate, amoxycillin trihydrate and cloxacillin sodium. The former have slightly higher levels of adsorption than the latter. A negative linear relationship was found between the amount of antibiotic adsorbed onto dry sorbitol and that originally added to sorbitol. When adsorption is expressed as the weight of drug adsorbed per unit weight of sorbitol, an 'apparent' Langmuir isotherm results. This suggests that there are a number of adsorption sites available for holding drug particles, these sites being different for the different antibiotics.

Adsorption↗
PubMed1993

Virulence of an Escherichia coli O157:H7 sorbitol-positive mutant.

Virulence and pathogenicity of an Escherichia coli O157:H7 sorbitol-positive mutant were investigated with an infant rabbit animal model as well as a battery of in vitro assays. Total cell lysate protein profiles, outer membrane protein profiles, plasmid profiles, and levels of cytotoxic activity against Vero cells were similar in the wild-type and mutant strains. Both adhered to intestinal epithelial cells in culture and reacted with fluorescein isothiocyanate-labeled antiserum against E. coli O157:H7. The mutant appeared to be similar to the wild type in all respects except in its ability to ferment sorbitol. [14C]sorbitol uptake and sorbitol-6-phosphate dehydrogenase activities were notably increased in the mutant strain. Diarrhea developed in rabbits administered the wild-type strain and in those fed the sorbitol-positive mutant. There was greater bacterial attachment and mucosal damage in the cecum and large intestine than in the small intestine. Scanning electron microscopy revealed bacteria adhering as single cells and as aggregates closely associated with mucus. Mucosal lesions consisted of areas of tissue necrosis with sloughing of epithelial cells. By transmission electron microscopy, electron-dense necrotic epithelial cells were visible in areas where bacteria were present, and epithelial cell debris containing bacteria was observed between the villar luminal surfaces. Light microscopy of epithelial cells of intestinal sections of infected rabbits revealed noticeable vacuolation and spherical, pyknotic nuclei. These data indicate that the sorbitol-negative phenotype is not associated with the pathogenicity of E. coli O157:H7.

Animals↗
PubMed1982

Effect of intracytoplasmic membrane development on oxidation of sorbitol and other polyols by Gluconobacter oxydans.

By using membrane-bound dehydrogenases, Gluconobacter oxydans characteristically accomplishes single-step oxidation of many polyols and quantitative release of the oxidation product into the medium. These cells typically differentiate by forming intracytoplasmic membranes (ICM) after exponential growth on glycerol. Earlier experiments demonstrated that glycerol-grown cells containing ICM oxidized glycerol more rapidly than cells which were harvested during exponential growth and lacked ICM (Claus et al., J. Bacteriol. 123:1169-1183). This report demonstrates that ICM are also formed after growth on sorbitol. Sorbitol-grown, ICM-containing maximum stationary-phase (MSP) cells showed from 50 to 300% greater oxidation (respiration) rates on mannitol, glycerol, glucose, meso-erythritol, and meso-inositol than did exponential-phase (EXP) cells which lacked ICM. Both EXP and MSP cells exhibited maximum sorbitol oxidation at pH 5.0, 38 degrees C, and 5% (wt/vol) sorbitol. When assayed under these optimum conditions, ICM-containing MSP cells demonstrated a 72% increase in respiration on sorbitol compared with that of EXP cells lacking ICM (oxygen quotients of 3,100 and 1,800, respectively). Gas chromatographic studies showed that sorbose was the only detectable product released from cells during oxygen quotient analysis. The specific activity of particulate-bound sorbitol dehydrogenase from ICM-containing MSP cells was twice that obtained from particulate fractions prepared from EXP cells lacking ICM. These results show that neither ICM formation after exponential growth nor increased respiration of other polyols is dependent upon the polyol used to grow cells. Our results suggest that increased respiratory activity of MSP cells is caused both by ICM formation and by increased synthesis (or activity) of the polyol dehydrogenases found in these membranes.

Glucose↗
PubMed1985

Agar media that indicate acid production from sorbitol by oral microorganisms.

Two varieties of agar medium (Trypticase [BBL Microbiology Systems]-serum-sorbitol-bromcresol purple agar [TSSB] and Trypticase-blood-sorbitol-CaCO3 agar [TBSCa]) indicating microbial acid production from sorbitol were tested. The media were devised for use in studies on the prevalence of sorbitol-fermenting human oral microorganisms incubated in an anaerobic or microaerophilic atmosphere containing 5 to 6% CO2. TSSB contains bromcresol purple as the pH indicator and NaHCO3 as the main buffering salt. TBSCa contains CaCO3 as both the buffering salt and the indicator of acid production. The growth yield of pure cultures of oral microorganisms on TBSCa was shown to equal that on blood agar incubated under similar conditions. TSSB inhibited the growth of several bacteria to various extents. The recovery of sorbitol-fermenting microorganisms from oral specimens was the greatest when the specimens were assayed with TBSCa. The poorer results obtained with TSSB were mainly due to the decoloration of the pH indicator in this medium and the presence of greater numbers of sorbitol false-positive colonies.

Acids↗
PubMed1990

In vivo osmoregulation of aldose reductase mRNA, protein, and sorbitol in renal medulla.

Sorbitol accumulates in renal medullary cells by synthesis from glucose in a reaction catalyzed by aldose reductase. Medullary sodium and urea are high and vary with urinary concentration. Sorbitol varies similarly, consistent with its role as a compatible intracellular organic osmolyte. We measured renal medullary sodium, urea, sorbitol, aldose reductase (protein and activity), and aldose reductase mRNA in rats treated to change medullary sodium and urea. In untreated Brattleboro rats all measurements were low and increased after 7 days of treatment with arginine vasopressin. In contrast, when normal rats were water deprived for 3 days, urea increased out of proportion to sodium, and sorbitol, aldose reductase, and aldose reductase mRNA were unchanged. After 2 h of diuresis, normal rats had lower medullary sodium and urea and reduced mRNA and sorbitol; however aldose reductase did not change. These data are consistent with previous results from cultured cells in which altered extracellular sodium, but not urea, leads to rapid changes in aldose reductase mRNA and slow changes (days) in aldose reductase. In addition, acute decreases in extracellular sodium increase leakage of sorbitol from cells. We also confirm previous results showing medullary glycerophosphorylcholine correlates best with urea, whereas the sum of all compatible osmolytes correlates best with sodium.

Aldehyde Reductase↗
PubMed1998

Regulation of sorbitol content in cultured porcine urinary bladder epithelial cells.

Sorbitol content was determined in porcine urinary bladder epithelial cells immediately after death of the animals and after primary culture of the cells at different osmolalities. In both instances, sorbitol content increased with urine and medium osmolality, respectively. For example, at 300 mosmol/kg the cultured cells contained 0.84 +/- 0.02 nmol/mg protein, at 600 mosmol/kg contained 21.7 +/- 0.95 nmol/mg protein, and at 900 mosmol/kg contained 59.5 +/- 2.8 nmol/mg protein. Similarly, aldose reductase activity rose from 0.27 +/- 0.04 mumol.h-1.mg protein-1 at 300 mosmol/kg to 1.81 +/- 0.16 at 600 mosmol/kg and to 3.02 +/- 0.33 at 900 mosmol/kg. These changes were, however, only observed when NaCl but not when urea was used to augment the medium osmolality, since urea equilibrated across the cell membrane. In contrast, sorbitol release from cells cultured at 900 mosmol/kg was slowest into a 900 mosmol/kg medium and fastest into a 300 mosmol/kg medium (63 +/- 16 nmol/10 min compared with 389 +/- 52 nmol/10 min). These studies demonstrate that the sorbitol content of porcine urinary bladder epithelium is regulated by changes both in sorbitol synthesis and sorbitol release. Thus the regulatory mechanisms in the urinary bladder seem to be similar to those present in the embryological related collecting duct.

Aldehyde Reductase↗
PubMed1986

Effects of diabetes and insulin treatment on sorbitol and water of rat lenses.

Since increased lens sorbitol and osmotic swelling are central causative features of cataract in diabetic rats, the effects of insulin treatment on lens sorbitol, water, sodium, and potassium were studied. The sorbitol concentration in early stage diabetic lenses was greater than in normal ones by 83 mmol/kg water, and the lens water was greater by 1.3%. Sodium was greater by 9 mmol/kg water; potassium was less by the same amount so that the sum of sodium and potassium was not different. In insulin-treated diabetic lenses, the sorbitol was less than in untreated diabetic lenses by 39 mmol/kg water, and the lens water was not different. Insulin restored the potassium, but not the sodium, to normal concentration so that the sum of sodium and potassium was greater by 16 mmol/kg water. The differences in lens water were less than would be expected on the basis of osmosis due to differences in sorbitol and suggested that the lenses were able to maintain their water content within a narrow range by losing or gaining solutes to offset the differences in sorbitol.

Alloxan↗
PubMed1990

Sorbitol adaptation of dental plaque in people with low and normal salivary-secretion rates.

Caries-related bacteriological and biochemical factors were studied in 12 persons with low and 11 persons with normal salivary-secretion rates before and after a four-week period of frequent mouthrinses with 10% sorbitol solution (adaptation period). After the adaptation period, an increase in total numbers of sorbitol-fermenting bacteria and mutans streptococci was found in plaque samples from both groups. The pH drop in dental plaque in situ after a mouthrinse with sorbitol was more pronounced after the adaptation period, especially in the group with the low salivary-secretion rate. Increased acid production in plaque from sorbitol was also observed in the two groups in vitro. The composition of the fermentation end-products was affected by the sorbitol exposure. It is suggested that in subjects with diminished natural protection against caries development, such as patients with low salivary-secretion rate, a cariogenic potential of sorbitol should be considered when this polyol is included in products that might be used frequently.

Adaptation, Physiological↗
PubMed1990

[Erythrocyte sorbitol levels in patients with thyrotoxicosis].

Hyperglycemia and impaired glucose tolerance are frequently observed in patients with hyperthyroidism. However, little is known about whether altered polyol metabolism in hyperthyroidism is present or not. To examine changes in polyol metabolism in hyperthyroidism, we investigated changes in erythrocyte sorbitol, glyceraldehyde reductase (GAR) and sorbitol dehydrogenase (SDH) activities during hyperthyroid and euthyroid states in patients with thyrotoxicosis. Mean levels of erythrocyte sorbitol and GAR were 32.0 +/- 1.6nM/g.Hb and 147.1 +/- 0.3mU/g.Hb, respectively. In thyrotoxic patients in a hyperthyroid state, these values were significantly higher than those in euthyroid controls. Mean level of erythrocyte SDH in thyrotoxic patients was weak but was significantly increased in comparison with that of euthyroid controls. However, mean levels of erythrocyte sorbitol and GAR were remarkably reduced to 23.6 +/- 1.4nM/g.Hb and 125.3 +/- 4.6mU/g.Hb in thyrotoxic patients in a euthyroid state after treatment with anti-thyroid drugs or by subtotal thyroidectomy. Mean level of SDH, on the other hand, was increased after the treatment. In addition, positive correlations were observed between the level of erythrocyte sorbitol or GAR, and the level of free thyroxine(FT4) or free triiodothyronine(FT3). A negative correlation was observed between the level of erythrocyte SDH and the level of FT4 or FT3. These results suggest that the level of erythrocyte sorbitol may be increased through direct acceleration of erythrocyte GAR activity by increased thyroid hormone levels in patients with thyrotoxicosis.

Adult↗