Inhibitory effects of oolong tea polyphenols on pancreatic lipase in vitro

Fifty-four polyphenols isolated from tea leaves were evaluated for their inhibitory activities against pancreatic lipase, the key enzyme of lipid absorption in the gut. (-)-Epigallocatechin 3-O-gallate (EGCG), which is one of major polyphenols in green tea, showed lipase inhibition with an IC50 of 0.349 microM. Moreover, flavan-3-ol digallate esters, such as (-)-epigallocatechin-3,5-digallate, showed higher activities of inhibition on lipase with an IC50 of 0.098 microM. On the other hand, nonesterified flavan-3-ols, such as (+)-catechin, (-)-epicatechin, (+)-gallocatechin, and (-)-epigallocatechin, showed zero and/or the lowest activities against pancreatic lipase (IC50 > 20 microM). These data suggested that the presence of galloyl moieties within the structure was required for enhancement of pancreatic lipase inhibition. It is well-known that flavan-3-ols are polymerized by polyphenol oxidase and/or heating in a manufacturing process of oolong tea. Oolonghomobisflavans A and B and oolongtheanin 3'-O-gallate, which are typical in oolong tea leaves, showed strong inhibitory activities with IC50 values of 0.048, 0.108, and 0.068 microM, respectively, even higher than that of EGCG. The oolong tea polymerized polyphenols (OTPP) were prepared for the assay from oolong tea extract, from which the preparation effectively subtracted the zero and/or less-active monomeric flavan-3-ols by preparative high-performance liquid chromatography. The weight-average molecular weight (Mw) and number-average molecular-weight (Mn) values of OTPP were 2017 and 903, respectively, by using gel permeation choromatography. OTPP showed a 5-fold stronger inhibition against pancreatic lipase (IC50 = 0.28 microg/mL) by comparison with that of the tannase-treated OTPP (IC50 = 1.38 microg/mL). These data suggested that the presence of galloyl moieties within their chemical structures and/or the polymerization of flavan-3-ols were required for enhancement of pancreatic lipase inhibition.     

The effects of human disturbance on a stingless bee community in a tropical rainforest

Human disturbance of tropical rainforests may change pollinator communities indirectly as a result of changes in resource availability. We studied the mechanisms by which human disturbance affects a community of major pollinators, stingless bees, in Sarawak, Malaysia. We surveyed forest structure and flowering activity, and conducted a nest census and a bait-trap survey of stingless bees, both in primary forests and in forests disturbed by logging and shifting cultivation. The densities of late successional trees and large trees (diameter at breast height >50 cm) were higher in the primary forests than in the disturbed forests. The density of flowering trees was lower one year after logging, but recovered in old disturbed forests because of the active flowering of pioneer trees that became established after the disturbance. Stingless bees nest only in large trees, and nest density was positively correlated with the density of large trees. However, we found no relationship between the numbers of foragers and floristic parameters. Some species preferred nesting in dipterocarps, while others preferred dead trees. The results of the bait-trap survey also indicated that some species were abundant in the primary forests, whereas other species were abundant in the disturbed forests. These results suggest that human disturbance alters the species composition of the stingless bee community. Such changes in the bee community may affect the reproductive success of plants, and ultimately forest composition.     

Angiotensin I-converting enzyme-inhibitory peptides obtained from chicken collagen hydrolysate

In this study, collagen extracted from chicken legs (which are the yellow keratin parts containing a nail) was hydrolyzed with various enzymes, and the angiotensin I-converting enzyme (ACE)-inhibitory activity of each hydrolysate was determined. The hydrolysate by treatment with an Aspergillus species-derived enzyme had the highest activity (IC 50 = 260 microg/mL). The fraction of this hydrolysate obtained by ultrafiltration with a molecular-weight cutoff of 3000 Da (low fraction) had a stronger activity (IC 50 = 130 microg/mL) than the fractionated one. This fraction was further fractionated by HPLC, and the peptides in the fraction with high ACE-inhibitory activity were identified. The amino acid sequences of the four peptides were identified using a protein sequencer. These peptides were synthesized to confirm their ACE-inhibitory activities; this showed that peptides with a Gly-Ala-Hyp-Gly-Leu-Hyp-Gly-Pro sequence had the highest activity (IC 50 = 29 microM). When the low fraction was administered to spontaneous hypertensive rats, a decrease in their blood pressure was observed after 2 h of administration, and a significant decrease in blood pressure (-50 mmHg) was observed after 6 h. Moreover, long-term administration studies indicated that the low fraction showed a significant suppression of increased blood pressure.     

Studies on absorption and hydrolysis of ethyl alpha-D-glucoside in rat intestine

Ethyl alpha-D-glucoside (alpha-EG) is normally contained in Sake, which has been taken by Japanese people since ancient times. In this study, the intestinal absorption of alpha-EG was investigated using rat everted intestinal sac. Furthermore, the alpha-EG hydrolytic activity in rat intestine was compared with disaccharides hydrolytic activities, and the effects of alpha-EG on disaccharides hydrolysis were examined using crude enzyme preparation from rat intestinal acetone powder. Glucose liberated from alpha-EG was detected in a serosal solution of everted rat intestinal sac, but it was only less than 4% of absorbed intact alpha-EG. alpha-EG absorption into small intestinal tissue was reduced by elimination of sodium ion from the mucosal solution or under the presence of phlorizin. The hydrolytic activity for alpha-EG was detected in crude enzyme preparation from rat intestinal acetone powder, but it showed a low value as compared to those for disaccharides. alpha-EG showed mixed type inhibition for maltose and sucrose hydrolysis, but inhibitory concentrations of alpha-EG required for 50% inhibition for the maltose and sucrose hydrolysis were higher than those of arabinose and acarbose. In conclusion, a small amount of alpha-EG was hydrolyzed and most of it was absorbed via SGLT1 as an intact form in the rat small intestine, and the inhibitory effect of alpha-EG on disaccharides hydrolysis was weak.     

Preparation of antimicrobial reduced lysozyme compatible in food applications

The structural and antimicrobial functions of lysozyme reduced with food-compatible reducing agents-cysteine (Cys) and glutathione (GSH)-were investigated. The disulfide bonds were partially reduced by thiol-disulfide exchange reactions under heat-induced denaturing conditions from 55 to 90 degrees C. The results showed that treatment of lysozyme with Cys and GSH resulted in the introduction of new half-cystine residues (2-3 residues/mol of protein). The released SH groups, in turn, rendered the lysozyme molecule more flexible, being accompanied by a dramatic increase in the surface hydrophobicity and exposure of tryptophan residues. As a consequence, the resulting reduced lysozymes were more capable of binding to lipopolysaccharides (LPS) and permeabilizing the bacterial outer membrane, as evidenced by the liposome leakage experiment, than were native or heated lysozyme. Both reduced lysozymes displayed significantly higher antimicrobial activity than native or heated lysozyme against Salmonella enteritidis (SE) in sodium phosphate buffer (10 mM, pH 7.2) at 30 degrees C for 1 h. Their minimal inhibitory concentrations (MICs) against the tested bacteria were about 150- and 25-fold lower than their respective MICs of native or heated lysozyme. The results suggest that partially reduced lysozyme could be used as a potential antimicrobial agent for prevention of SE attack.     

Enantioseparation of isoxanthohumol in beer by hydroxypropyl-gamma-cyclodextrin-modified micellar electrokinetic chromatography

Chiral resolution of isoxanthohumol (IX) in beer samples was performed by hydroxypropyl-gamma-cyclodextrin-modified micellar electrokinetic chromatography. The optimum running conditions were found to be 20 mM phosphate buffer (pH 7.0) containing 45 mM hydroxypropyl-gamma-cyclodextrin and 100 mM sodium dodecyl sulfate with an effective voltage of +20 kV at 20 degrees C using direct detection at 210, 295, and 370 nm. IX was detected in 12 beer samples and the total levels of (+)- and (-)-IX ranged from 0.15 to 1.4 mg/L. But the amount of xanthohumol (XN) was below the detection limit (0.017 mg/L). Each level of (-)-IX was almost the same as that of (+)-IX, suggesting that IX was a racemic mixture in these beer samples. The racemization of IX in beer could be attributed to the production of a racemic mixture from XN during boiling and to the fact that IX enantiomers were easily interconverted.     

Quality Characteristics of Waxy Hexaploid Wheat (Triticum aestivum L.): Properties of Starch Gelatinization and Retrogradation

The viscoelastic properties and molecular structure of the starch isolated from waxy (amylose-free) hexaploid wheat (WHW) (Triticum aestivum L.) were examined. WHW starch generally had lower gelatinization onset temperature, peak viscosity, and setback than the starch isolated from normal hexaploid wheat (NHW). Differential scanning calorimetry (DSC) showed that WHW starch had higher transition temperatures (T_o, T_p, and T_c) and enthalpy (ΔH) than NHW starch. However, when compared on the basis of amylopectin (AP) content, ΔH of WHW starch was almost statistically identical to that of its parental varieties. Typical A-type X-ray diffraction patterns were observed for the starches of WHW and its parental varieties. Somewhat higher crystallinity was indicated for WHW starch. WHW starch was also characterized by having greater retrogradation resistance. The high-performance size-exclusion chromatography (HPSEC) of amylopectin showed that each amylopectin yielded two fractions after debranching. Although WHW amylopectin had somewhat long B chains, little difference was observed in the ratio of Fr.III/ Fr.II between WHW and its parental varieties.     

Hot spring bathing accelerates wound healing and enhances heat retention effect in guinea pigs

This study aimed to demonstrate the effects of hot springs on wound healing and heat retention by performing comparative experiments with tap water. The hot spring water used in this study was from an alkaline hot spring that was rich in sodium and chloride ions and exhibited high reducibility. Guinea pigs were divided into a hot spring bathing group and a tap water bathing group, and a bathing test was conducted for eight consecutive days. A comparison of the plasma amino acid composition between the two groups after the bathing test revealed differences in the concentrations of several amino acids associated with wound healing. Image analysis demonstrated that wounds made on the abdominal skin of guinea pigs were significantly contracted by hot spring bathing compared to that by tap water bathing, and histopathological findings showed that wound healing was accelerated. In the thermography test, changes in body surface temperature after bathing were investigated in both groups. The heat retention effect was not observed in the tap water bathing group after bathing, whereas it was enhanced in the hot spring bathing group until 30 min after bathing. In conclusion, this study demonstrated that hot spring bathing accelerates wound healing and has a more significant heat retention effect than tap water bathing.     

Harvest index is a critical factor influencing the grain yield of diverse wheat species under rain-fed conditions in the Mediterranean zone of southeastern Turkey and northern Syria

Environmental and plant factors critical to the grain yields of bread (Triticum aestivum L.), durum (T. durum L.) and emmer (T. dicoccum L.) wheat cultivars were investigated at two Mediterranean rain-fed field sites: Adana in southeastern Turkey (2009 and 2010) and Aleppo in northern Syria (2009). The grain yield (GY) and biological yield (BY) of most cultivars were higher in Adana than in Aleppo, and the lower GY in Aleppo resulted from lower harvest index (HI) and lower BY due to higher temperatures and lower rainfall. The variations in the HI among cultivars were greater in Adana than in Aleppo. The GY was closely related to the HI but not the BY across cultivars at each site, and a higher GY was accompanied by a superior conversion-efficiency of incident radiation during the grain filling period for grain yield [GY/Ra, where Ra is the cumulative radiation for 30 days after heading (D₃₀)] across all observations. The GY/Ra correlated negatively with the average temperature for D₃₀, and higher HI values resulted in higher GY/Ra. In Adana, the time from anthesis to physiological-maturity decreased as the average temperature for D₃₀ increased, resulting in a lower HI. Cultivars exhibiting the early heading trait can effectively escape the negative impacts of terminal high-temperature and water-shortage conditions on the HI. The results suggested that the HI is a critical factor for GY across diverse wheat cultivars under terminal high-temperatures and water-shortages in Mediterranean areas, and the BY is also an important factor under severe water-limitation conditions.