Effect of a sports drink based on highly-branched cyclic dextrin on cytokine responses to exhaustive endurance exercise.

BACKGROUND: Aim of the present study was to compare the effects of highly branched cyclic dextrin (HBCD) drink with a glucose-based control drink on immunoendocrine responses to endurance exercise. METHODS: Using a randomized, double-blind placebo-controlled cross-over design, seven male triathletes participated in two duathlon races separated by one month, consisting of 5 km of running, 40 km of cycling and 5 km of running. In the first race, four athletes consumed the HBCD-based drink and three athletes consumed the glucose-based drink. In the second race, three athletes consumed the HBCD-based drink and four athletes consumed the glucose-based drink. We collected blood and urine samples before and after the races to analyze leukocyte count and concentrations of hormones and cytokines. RESULTS: Lymphocyte and neutrophil counts increased significantly after exercise in both trials (P<0.05), but were not significantly different between the trials. Plasma noradrenalin concentration increased significantly (P<0.05) during exercise in the glucose trial, but not in the HBCD trial. Plasma concentrations of interleukin (IL)-8 and IL-10 increased significantly during exercise in both trials (P<0.05) but were not significantly different between the trials. Post-race urinary IL-8, IL-10 and IL-12p40 concentrations were significantly lower in the HBCD trial compared with the glucose trial (P<0.05), although the plasma concentrations of these cytokines were not significantly different between both trials. CONCLUSION: These results suggest that the HBCD-based drink may attenuate the stress hormone response, and reduce the urinary cytokine levels following exhaustive exercise.

Evaluation of the distribution and orientation of remineralized enamel crystallites in subsurface lesions by X-ray diffraction.

Remineralization is the process by which hydroxyapatite (HAp) is restored in enamel subsurface lesions, and transversal microradiography (TMR) has been used to analyze remineralization in terms of the recovery of mineral content. In this study, we directly detected the distribution and orientation of longitudinal HAp crystallite at the remineralized zone in enamel subsurface lesions by using an X-ray microbeam (6-mum diameter) diffraction method. This method was demonstrated and involves the simultaneous detection of wide-angle X-ray diffraction (WAXRD) and small-angle X-ray scattering (SAXS). WAXRD reflects the amount of HAp crystallites, and SAXS reflects that of voids in crystallites. The polished surface of a bovine enamel block was divided into three zones of sound, demineralized, and remineralized zones. Thin sections of approximately 150 mum thickness were then cut perpendicular to the surface, and subjected to WAXRD and SAXS following TMR. The increase in the amount of HAp crystallites and the decrease in voids in the crystallites at the remineralized zone were detected by WAXRD and SAXS, respectively, which was consistent with the result of TMR. This study indicates that both the spatial distribution and orientation of the restored HAp crystals in the remineralization process at the subsurface lesion can be simultaneously analyzed by the X-ray diffraction methods.

Long-term survival of a patient with type A thymoma and Masaoka's stage IV b: case report.

Thymoma is the most common neoplasm in the anterior mediastinum. This report presents an extremely rare case of a type A thymoma with Masaoka's stage IV b due to lymph node metastasis. In 1997, a 59-year-old man underwent complete resection of a stage IV b type A thymoma with postoperative radiotherapy to the mediastinum. In 2006, small nodules were detected in the anterior mediastinum and above the right diaphragm and tumor resection was performed. The two lesions were both histologically diagnosed as recurrences of the type A thymoma. There has been no evidence of recurrence 15 months after the second surgery.

Psychological stress-reducing effect of chocolate enriched with gamma-aminobutyric acid (GABA) in humans: assessment of stress using heart rate variability and salivary chromogranin A.

We studied the psychological stress-reducing effect of chocolate enriched with gamma-aminobutyric acid (GABA), on stress induced by an arithmetic task using changes of heart rate variability (HRV) and salivary chromogranin A (CgA). Subjects ingested 10 g chocolate enriched with 28 mg GABA (GABA chocolate); 15 min after the ingestion, subjects were assigned an arithmetic task for 15 min. After the task, an electrocardiogram was recorded and saliva samples were collected. HRV was determined from the electrocardiogram, and the activity of the autonomic nervous system was estimated through HRV. The CgA concentration of all saliva samples, an index for acute psychological stress, was measured. From HRV, those taking GABA chocolate made a quick recovery to the normal state from the stressful state. The CgA value after the task in those taking GABA chocolate did not increased in comparison with that before ingestion. From these results, GABA chocolate was considered to have a psychological stress-reducing effect.

Evaluation of enamel crystallites in subsurface lesion by microbeam X-ray diffraction.

Early caries lesion is a demineralization process that takes place in the top 0.1 mm layer of tooth enamel. In this study, X-ray microbeam diffraction was used to evaluate the hydroxyapatite crystallites in the subsurface lesion of a bovine enamel section and the results are compared with those obtained by transversal microradiography, a method commonly used for evaluation of tooth mineral. Synchrotron radiation from SPring-8 was used to obtain a microbeam with a diameter of 6 microm. Wide-angle X-ray diffraction reports the amount of hydroxyapatite crystals, and small-angle X-ray scattering reports that of voids in crystallites. All three methods showed a marked decrease in the enamel density in the subsurface region after demineralization. As these diffraction methods provide structural information in the nanometre range, they are useful for investigating the mechanism of the mineral loss in early caries lesion at a nanometre level.

[Pulmonary granuloma possibly caused by a fish bone material].

We herein describe an extremely rare case of pulmonary granuloma possibly caused by a fish bone material. A 60-year-old woman with hemosputum was found to have a right pulmonary nodule. Chest computed tomography showed a nodule with pleural retraction, vascular convergence and ground glass opacity, measuring 23 mm in diameter, located at the S2 Of right upper lobe. Based on these findings, this nodule was considered to be a primary lung cancer, and pulmonary resection was performed. Macroscopically, a foreign material was found in the lesion and the pathologic diagnosis of the nodule was foreign body granuloma possibly due to a fish bone material.

Fluids containing a highly branched cyclic dextrin influence the gastric emptying rate.

The rates of gastric emptying for highly branched cyclic dextrin (HBCD) and other carbohydrate (CHO) solutions were examined using ultrasonograph techniques. Ten healthy volunteers ingested water, physiological saline, or solutions containing various CHO, such as HBCD, glucose, maltose, sucrose, and commercially available dextrin. After a subject drank one of the solutions, the relaxed cross-sectional area of the pylorus antrum was measured at rest by real-time ultrasonography. The time required for gastric emptying was correlated with the relaxed cross-sectional area of the pylorus antrum. Among all of the solutions tested, physiological saline was transferred fastest from the stomach to the small intestine. For solutions of the same CHO, 5 % solution was transferred faster than 10 % solution. For CHO solutions other than HBCD, a low osmotic pressure was associated with rapid transfer from the stomach. The gastric emptying time (GET) of HBCD solution increased with an increase in its concentration. A shorter GET was observed for the CHO solutions at 59 to 160 mOsm regardless of their concentration. A sports drink based on 10 % HBCD adjusted to 150 mOsm by the addition of various minerals, vitamins, and organic acids was evacuated significantly (p < 0.05) faster than a 10 % HBCD solution or a sports drink based on 10 % commercially available dextrin (DE16), which has a higher osmotic pressure (269 mOsm). Our results suggest that a shorter GET could be achieved with CHO solutions with osmotic pressures of 59 - 160 mOsm. Therefore, a sports drink based on 10 % HBCD adjusted to 150 mOsm by the addition of minerals, vitamins, and organic acids could supply adequate quantities of CHO, fluid, and minerals simultaneously in a short time, without increasing GET.

Antidiabetic effect of glycyrrhizin in genetically diabetic KK-Ay mice.

We, previously demonstrated that one shot administration of glycyrrhizin (Grz) reduced the postprandial blood glucose rise, using Std ddY mice. Subsequently, we evaluated the effects of long-term Grz treatment (2.7, 4.1 g/kg diet) on diabetic symptoms using genetically non-insulin dependent diabetic model mice (KK-Ay). Male KK-Ay mice were divided into 3 groups: the control group, 0.27% Grz diet (2.7 g of Grz/kg diet) group and 0.41% Grz diet (4.1 g of Grz/kg diet) group. The elevation of blood glucose concentration was almost entirely suppressed in mice fed the 0.41% Grz diet 7 weeks after the beginning of test feeding, although it was not suppressed in mice fed the control diet or the 0.27% Grz diet. Water intake in the control and 0.27% Grz diet groups increased gradually, whereas, this was not true in the 0.41% Grz diet group. Grz treatment significantly lowered blood insulin level. Throughout the experiment, Grz did not affect the food intake or body weight among the three groups. The mice fed the 0.41% Grz diet also improved their tolerance to oral glucose loading 9 weeks after the beginning of test feeding. This study shows that Grz has an antidiabetic effect in noninsulin-dependent diabetes model mice.

Practical production of (S)-1,2-propanediol and its derivative through baker's yeast-mediated reduction.

The enantiomeric excess (ee) of (S)-1,2-propanediol produced by baker's yeast-mediated reduction of 1-acetoxy-2-propanone was improved to 96% ee by a fed-batch operation of the substrate. A similar reduction of 1-benzoyloxy-2-propanone stopped because of the inhibition toward the enzymes for NADPH regeneration by the reduction product. The inhibition was prevented using resin that adsorbs the product from the reaction mixture, and 70 g/l substrate was reduced yielding (S)-1-benzoyloxy-2-propanol at > 99% ee.

Enhancement of swimming endurance in mice by highly branched cyclic dextrin.

We investigated the ergogenic effect in mice of administering highly branched cyclic dextrin (HBCD), a new type of glucose polymer, on the swimming endurance in an adjustable-current swimming pool. Male Std ddY mice were administered a HBCD, a glucose solution or water via a stomach sonde 10 min before, 10 min after or 30 min after beginning swimming exercise, and were then obliged to swim in the pool. The total swimming period until exhaustion, an index of the swimming endurance, was measured. An ergogenic effect of HBCD was observed at a dose of 500 mg/kg of body weight, whereas it had no effect at a dose of 166 mg/kg of body wt (p < 0.05). The mice administered with the HBCD solution 10 min after starting the exercise were able to swim significantly longer (p < 0.05) than the mice who had ingested water or the glucose solution. The rise in mean blood glucose level in the mice administered with HBCD, which was measured 20 min after starting swimming, was significantly lower (p < 0.05) than that in the mice administered with glucose, although it was significantly higher (p < 0.05) than that in the mice administered with water. The mean blood insulin rise in the mice given HBCD was significantly lower (p < 0.05) than that in the mice given glucose. The mice administered with HBCD 30 min after starting the exercise swam significantly longer (p < 0.05) than the mice who had ingested water, although the enhancement of swimming time was similar to that of the glucose-ingesting mice. The gastric emptying rate of the HBCD solution was significantly faster (p < 0.05) than that of the glucose solution. However, this glucose polymer must have spent more time being absorbed because it has to be hydrolyzed before absorption, reflecting a lower and possibly longer-lasting blood glucose level. We conclude that the prolongation of swimming endurance in mice administered with HBCD depended on its rapid and longer-lasting ability for supplying glucose with a lower postprandial blood insulin response, leading to a delayed onset of fatigue.

Permeation of hesperidin glycosides across Caco-2 cell monolayers via the paracellular pathway.

The intestinal permeability to hesperidin glycosides was investigated by using a cultured monolayer of Caco-2 as a model for the small intestinal epithelium. Hesperidin glycosides were added to the apical side of the monolayer, and the substances that permeated to the basolateral side were determined by HPLC. Whereas hesperidin did not permeate across the Caco-2 monolayer, probably owing to its low solubility, the hesperidin glycosides did permeate. The transepithelial transport of hesperidin glycosides occurred in time- and dose-dependent manners. The transport was observed to be energy-independent, and was inversely correlated with the transepithelial electrical resistance (TEER) of the monolayer. These results suggest that hesperidin glycosides permeate across the Caco-2 cell monolayer via the paracellular pathway.

Formation of Inclusion Complexes of Cycldextrin with Ethanol under Anhydrous Conditions.

Complex formation of poorly water soluble organic compounds with cyclodextrin (CD) is quite difficult in an aqueous cyclodextrin system. Formation of the inclusion complex of d-limonene, phenyl ethanol, acetophenone, or menthol was investigated in a slurry form of α-, ß-, or γ-CD in organic solvents or alcohol under anhydrous conditions. Ethanol and methanol were found to be good solvents for this method. The use of ethanol as the solvent was investigated in greater detail. There existed an optimal amount of ethanol for the maximum inclusion of d-limonene as the guest compound. However, an excess of ethanol inhibited the inclusion. An adsorption model of alcohol on CD, analogous to the substrate inhibition model of enzyme kinetics, could correlate the inclusion ratio with the amount of alcohol added to CD.

A novel acid phosphatase from Aspergillus niger KU-8 that specifically hydrolyzes C-6 phosphate groups of phosphoryl oligosaccharides.

We had analyzed the detailed structures of the phosphoryl oligosaccharide-1 (PO-1) fraction that was the main component of phosphoryl oligosaccharides (POs) prepared from a potato starch hydrolysate. PO-1 fraction was made up of 3-phosphoryl oligosaccharides and 6-phosphoryl oligosaccharides. Aspergillus niger strain KU-8 produced two types of intracellular acid phosphatase (EC 3.1.3.2, ACPase); ACPase I and II. ACPase II preferentially dephosphorylated 6-phosphoryl oligosaccharides rather than 3-phosphoryl oligosaccharides. The molecular weight of the enzyme was estimated as 66 kDa by SDS-polyacrylamide gel electrophoresis and about 260 kDa by gel filtration, implying the active form to be a tetramer. The optimum pH and temperature of the enzyme were 2.0-2.5 and 60 degrees C, respectively. ACPase II was stable below 50 degrees C for 30 min and pH 2.0-10.0 for 60 min. In spite of the strict specificity toward 6-phosphoryl oligosaccharides in the PO-1 fraction, ACPase II was able to hydrolyze Fru-1,6-di-P, ATP, pyrophosphate, and polyphosphate as well as pNPP and Glc-6-P, a broad substrate specificity.

Lowering effect of phenolic glycosides on the rise in postprandial glucose in mice.

Glycosides were screened for their lowering effect on the postprandial blood glucose rise in vivo. The effect of phlorizin and other phenolic glycosides on the postprandial blood glucose response to glucose ingestion was evaluated in Std ddY mice. When phlorizin was simultaneously added, the peak blood glucose level was significantly decreased by 51% (p < 0.01) compared to vehicles following glucose ingestion by mice, while the blood insulin responses were generally similar. Screening experiments were conducted with different classes of phenolic glycosides added to a glucose solution. Reductions of 40-52% (p < 0.05) were observed in vehicles containing arbutin, 4-hydroxyphenyl-alpha-D-glucopyranoside (hydroquinone-alpha-glucoside) or glycyrrhizin, and of only 15-31% (not significant) in vehicles containing neohesperidin dihydrochalcone, glycyrrhetinic acid monoglucuronide, or 3,4-dimethoxyphenyl-beta-D-glucopyranoside. No lowering effect was observed in vehicles containing salicin. Since glycyrrhizin, arbutin, and hydroquinone-alpha-glucoside blunted to varying degrees the postprandial blood glucose rise following glucose ingestion, they may be useful adjuvants for the treatment of diabetic subjects.

NADPH-dependent reduction of ethyl acetoacetate coupled with ethanol oxidation in Kloeckera magna.

We evaluated the catalytic ability of 29 yeast strains to reduce ethyl acetoacetate (EA) in the presence of ethanol or glucose. In 18 yeast strains, the reduction in the presence of ethanol proceeded as well as in the presence of glucose. Among them, Kloeckera magna (AKU 4704) effectively catalyzed the NADPH-dependent reduction of EA in the presence of ethanol. In this reduction, 1 mol of EA was reduced by consuming 1 mol of ethanol. We found that the NADPH regeneration system responsible for EA reduction in K. magna was coupled with oxidation of acetaldehyde to acetic acid catalyzed by an NADP(+)-dependent aldehyde dehydrogenase.

Acceptor specificity of cyclodextrin glucanotransferase from an alkalophilic Bacillus species and synthesis of glucosyl rhamnose.

Cyclodextrin glucanotransferase [1,4-alpha-D-glucan 4-alpha-D-(1,4-alpha-D-glucano)-transferase (cyclizing), EC 2. 4. 1. 19] from an alkalophilic Bacillus species A2-5a had a wider acceptor specificity than that from B. macerans, which was similar to those from B. stearothermophilus and B. circulans. Glucosyl rhamnose produced by the CGTase was identified as glucopyranosyl-alpha-1,4-rhamnopyranose by alpha- and beta-glucosidase treatments, and 1H- and 13C-NMR analyses.

Synthesis of neohesperidin glycosides and naringin glycosides by cyclodextrin glucanotransferase from an alkalophilic Bacillus species.

Cyclodextrin glucanotransferase from an alkalophilic Bacillus species produced neohesperidin monoglucoside and a series of its maltooligoglucosides by transglycosylation with neohesperidin as an acceptor and soluble starch as a donor. As the reaction using beta-CD as a donor at an alkaline pH was very effective for solubilizing neohesperidin, the amount of glycosides formed was increased. As a result, its amount with beta-CD at pH 10 was about 7 times greater than that with soluble starch at pH 5. Neohesperidin monoglucoside was purified from the reaction mixture by glucoamylase and naringinase treatments, an Amberlite XAD-16 column, a Sephadex LH20 column, and HPLC on an ODS column. The structure of the purified monoglucoside was identified as 3G-alpha-D-glucopyranosyl neohesperidin by FAB-MS, methylation analysis, and 1H- and 13C-NMR. The solubility of neohesperidin monoglucoside in water was approximately 1500 times higher than that of neohesperidin, and the bitterness of the monoglucoside was about 10 times less than that of neophesperidin. In addition, naringin was also glycosylated by the same method as neohesperidin, and its monoglucoside was identified as 3G-alpha-D-glucopyranosyl naringin. The solubility of naringin monoglucoside in water was also at least 1000 times higher than that of naringin without altering its bitterness.

Vitamin A is involved in estrogen-induced cell proliferation but not in cytodifferentiation of the chicken oviduct.

We examined vitamin A-deficient chicks to determine whether vitamin A affects the estrogen-induced development of the chick oviduct. When oviduct development was stimulated for 5 days with the synthetic estrogen, diethylstilbestrol, the wet weight of the oviduct in vitamin A-deficient chicks was only half that in control chicks. The DNA content in this tissue showed that the decreased oviduct weight in the vitamin A-deficient chicks was caused by the decreased proliferation of oviduct cells. However, the estrogen-induced expression of the ovalbumin gene was not affected by the vitamin A deficiency, suggesting that estrogen-induced cytodifferentiation is not affected by vitamin A. To clarify the vitamin A action on estrogen-induced development in the oviduct, transcripts of nuclear estrogen receptor (ER) and all-trans-retinoic acid (RAR alpha, beta and gamma) receptors, which exert the effects of estrogen and vitamin A, were measured. The ER, RAR alpha and RAR beta genes, but not that of RAR gamma, were expressed during oviduct development, indicating that estrogen and vitamin A may control the expression of target genes through their cognate receptors. Thus, we have shown that vitamin A is involved in estrogen-induced cell proliferation but not in cytodifferentiation of the chicken oviduct.

[Inhibitory effects of hydroquinone-alpha-glucoside on melanin synthesis].

Inhibitory effects of hydroquinone-alpha-glucoside (HQ-alpha-G) on the melanogenesis were investigated and compared with those of arbutin. The levels of inhibitory effects of HQ-alpha-G and arbutin on the tyrosinase activity were nearly the same. Inhibitory effects of both compounds on the melanogenesis, were studied using cultured B16 melanoma cells, and HQ-alpha-G was also found to have a similar effect to that of arbutin without inhibiting cell growth. In this experiment, while HQ-alpha-G hardly inhibited cell growth at 1 mM, arbutin inhibit it significantly at the same concentration. From these results it is suggested that HQ-alpha-G as well as arbutin inhibited the melanogenesis by affecting tyrosinase rather than by killing melanocytes. Furthermore, the melanogenesis of guinea-pigs with brown hair was reduced to about 80% by applying them each compound. The great differences in toxicity to normal human keratinocyte were not recognized between these two glucosides. It is, therefore, considered that HQ-alpha-G is an effective and safe ingredient for cosmetics.