This study examines the possibility of directly producing and utilizing useful substances in the intestines of animals using anaerobic bacteria that can grow in the intestines of animals. A facultative anaerobe producing a large amount of α-glucosidase inhibitor was isolated from hay and identified and named Bacillus coagulans CC. The main compound of α-glucosidase inhibitor produced by Bacillus coagulans CC was identified as 1-deoxynojirimycin. α-glucosidase inhibitor activity was confirmed in the intestinal contents and feces of mice orally administered with spores of this strain, and it was confirmed that this strain could efficiently reach the intestines, proliferate, and produce α-glucosidase inhibitors. As a result of administering Bacillus coagulans CC to mice at 109 cells per 1 kg body weight of spores for 8 weeks, the high-carbohydrate diet and the high-fat diet showed a 5% lower weight gain compared to the non-administrated group. At this point, in the spore-administered group, a decrease was observed in both the visceral and subcutaneous fat layers of the abdomen and thorax in both high-carbohydrate and high-fat diet groups compared to the non-administered group on computed tomography. The results of this study show that α-glucosidase inhibitors produced in the intestine by specific strains can work efficiently.
Polyamines have been reported to have cell proliferative and anti-inflammatory effects on normal metabolism in the body. This study aimed to investigate polyamine content of AIG01 pepper and the anti-inflammatory effect of AIG01 pepper extract (PAE) in mice. Polyamine content was analyzed by HPLC after acid hydrolysis of peppers with different acidic solvents. AIG01 pepper has the highest total polyamine content at about 1.5 mg/g. In LPS-stimulated RAW264.7, PAE inhibits nitric oxide production in a concentration-dependent manner and decreased the levels of pro-inflammatory cytokines. PAE has been shown to inhibit phosphorylation of MAPK/ERK. In TPA-stimulated Balb/C, PAE treatment showed tissue-level reductions in pro-inflammatory cytokines, reductions in ear thickness, and inhibition of neutrophil invasion. The polyamine content, polyamine extraction efficiency and anti-inflammatory effect of AIG01 obtained in this study suggest that it is useful as a raw material for the treatment of inflammatory diseases.
We selected Bacillus licheniformis NY1505 by screening a strain capable of producing α-glucosidase inhibitors in both aerobic and anaerobic environments in vitro and spore formation. To confirm whether this strain proliferates in the intestine and produces α-glucosidase inhibitor, the spores of this strain were administered to mice orally. As the results, it was confirmed that 107 cells and about 300 units of α-glucosidase inhibitor per 1 g feces were excreted in the feces after three weeks of administration as spores. And after two weeks of stopping administration, Bacillus licheniformis NY1505 in the intestine are cleared. This means that Bacillus licheniformis NY1505 steadily proliferated in the intestine and produced α-glucosidase inhibitors and excreted in the feces. Also, it has an advantage in its use as it can easily eliminate Bacillus licheniformis NY1505 from the intestine. This method of ingesting only microorganisms is a more efficient and new method than the existing method of administering an α-glucosidase inhibitor that consumes a large amount of purified product. This method shows a process in which microorganisms capable of proliferating in the intestine directly produce and supply specific secondary metabolites in the intestine.
We recently bred a cucumisin-containing Korean variety of melon. However, it is not known whether the cucumisin macromolecule can pass the intestinal barrier. We developed a polyclonal antibody against cucumisin and tried to confirm whether orally administered cucumisin passes through the intestinal tract with mouse. Cucumisin was isolated and purified from the improved Korean melon by ion exchange chromatography. Protein size was estimated by SDS-PAGE and protease activity was confirmed by performing zymography and fibrin plate assay. The purified cucumisin largely comprised a native form (67 kDa) and mature form (54 kDa), and displayed fibrinolytic activity, as evident by the decomposition of gelatin and fibrin to form a clear zone. To confirm the transition to blood following the oral administration of cucumisin, a polyclonal antibody against cucumisin was produced. Use of the antibody in ELISA and immunoblotting analyses confirmed the transfer of cucumisin to the blood of cucumisin-fed mice as compared to those not fed cucumisin. Orally administered cucumisin can migrate through the intestinal barrier and into the blood. The fibrinolytic activity of cucumisin purified from the improved Korean melon might be exploited for development as a raw material for a thrombolytic drug.
In this study, we used phenotypic and genetic analysis to investigate Double haploid (DH) lines derived from normal corn parents (HF1 and 11S6169). DH technology offers an array of advantages in maize genetics and breeding as follows: first, it significantly shortens the breeding cycle by development of completely homozygous lines in two or three generations; and second, it simplifies logistics, including requiring less time, labor, and financial resources for developing new DH lines compared with the conventional RIL population development process. In our study, we constructed a maize genetic linkage map using SSR markers and a DH population derived from a cross of normal corn (HF1) and normal corn (11S6169). The DH population used in this study was developed by the following methods: we crossed normal corn (HF1) and normal corn (11S6169), which are parent lines of a normal corn cultivar, in 2014; and the next year, the F1 hybrids were crossed with a tropicalized haploid inducer line (TAIL), which is homozygous for the dominant marker gene R1-nj (Nanda and Chase in Crop Sci 6:213–215, 1966), and we harvested seeds of the haploid lines. A total of 200 SSR markers were assigned to 10 linkage groups that spanned 1145.4 cM with an average genetic distance between markers of 5.7 cM. 68 SSR markers showed Mendelian segregation ratios in the DH population at a 5% significance threshold. A total of 15 quantitative trait loci (QTLs) for plant height (PH), ear height (EH), ear height ratio (ER), leaf length (LL), ear length (EL), set ear length (SEL), set ear ratio (SER), ear width (EW), 100 kernel weight (100 KW), and cob color (CC) were found in the 121 lines in the DH population. The results of this study may help to improve the detection and characterization of agronomic traits and provide great opportunities for maize breeders and researchers using a DH population in maize breeding programs.
Abstract Background Honey is a natural product used as food, medicine, or cosmetics for very long time and is made by bees. Honey contains various components such as sugar, protein, minerals, and vitamins. Honey is made by Apis cerana or Apis mellifera, which commonly has major royal jelly proteins (MRJPs) as a major protein. To discriminate between natural honey (NH) and artificial honey (AH), many researchers tried method of physicochemical analysis. However, the analysis results were ambiguous and not stable. Results We have produced a monoclonal antibody that recognizes MRJPs of honeys in common. Monoclonal antibody has advantage such as accuracy, sensitivity, and stability as the standard. The specificity and affinity of produced antibody were measured by western blotting and enzyme‐linked immunosorbent assay. As a result, this monoclonal antibody specifically recognized MRJPs of NH and did not recognize AH which has not including MRJPs. Conclusion Natural honey could be able to distinguish from AH accurately by using this monoclonal antibody. Also, this method could be commercially applicable.
Two new flavonoids, bismilachinone (11) and smilachinin (14), were isolated from the leaves of Smilax china L. together with 14 known compounds. Their structures were elucidated using spectroscopic methods. The PTP1B, alpha-glucosidase, and DPP-IV inhibitory activities of compounds 1-16 were evaluated at the molecular level. Among them, compounds 4, 7, and 10 showed moderate DPP-IV inhibitory activities with IC50 values of 20.81, 33.12, and 32.93 mu M, respectively. Compounds 3, 4, 6, 11, 12, and 16 showed strong PTP1B inhibitory activities, with respective IC50 values of 7.62, 10.80, 0.92, 2.68, 9.77, and 24.17 mu M compared with the IC50 value for the positive control (ursolic acid: IC50 = 1.21 mu M). Compounds 2-7, 11, 12, 15, and 16 showed potent alpha-glucosidase inhibitory activities, with respective IC50 values of 8.70, 81.66, 35.11, 35.92, 7.99, 26.28, 11.28, 62.68, 44.32, and 70.12 mu M. The positive control, acarbose, displayed an IC50 value of 175.84 mu M. In the kinetic study for the PTP1B enzyme, compounds 6, 11, and 12 displayed competitive inhibition with K-i values of 3.20, 8.56, and 5.86 mu M, respectively. Compounds 3, 4, and 16 showed noncompetitive inhibition with K-i values of 18.75, 5.95, and 22.86 mu M, respectively. Molecular docking study for the competitive inhibitors (6, 11, and 12) radically corroborates the binding affinities and inhibition of PTP1B enzymes. These results indicated that the leaves of Smilax china L. may contain compounds with anti-diabetic activity. (C) 2016 Elsevier Ireland Ltd. All rights reserved.
Capsaicinoids, the pungent component of chili peppers, are generally analyzed by precise analytical techniques, such as gas chromatography and high-performance liquid chromatography (HPLC), but these are not practical for the mass analyses of samples. To analyze mass samples rapidly, a colorimetric method was suggested. In this work, pigments and capsaicinoids were efficiently separated from chili pepper extract by sequential solid–liquid extraction and liquid–liquid extraction in test tubes followed by a colorimetric analysis on the capsaicinoids by a selective chromogenic reaction with Gibbs reagent (2,6-dichloroquinone-4-chloroimide). In the comparison of the capsaicinoid content by the colorimetric method and HPLC using acetone extracts of fresh pepper and dry red pepper as samples, R2 was 0.9973 and 0.9816, respectively, which shows a high linear correlation. In addition, a minimum of 1 μg/mL capsaicinoids can be detected and it was therefore determined that the method can efficiently analyze a great quantity of samples in a short time.
Ethnopharmacological relevance: The dried rhizome of Coptis chinensis Franch. (Huanglian) has been widely used in Asian traditional medicine. It was already known that Coptis chinensis Franch. rhizome has various pharmacological properties including its anti-oxidant, anti-cancer, and anti-inflammatory activity.Aim of the study: To evaluate the immune-enhancement effect of the Coptis chinensis Franch. rhizome extract on helper T cells and its signaling mechanism.Materials and methods: MOLT-4 human T cell line was used to investigate the effect of the Coptis chinensis Franch. rhizome extract. Cell viability was measured by the MTT assay and cytokine expression level was analyzed by ELISA and qRTPCR. MAPKs signal molecule's activation level was detected by immunoblotting.Results: The expression of IFN-gamma, a cytokine of type I helper T (Th1) cell, increased; however, IL-4 was not affected by the Coptis chinensis Franch. rhizome extract. Other Th1 cytokines, such as IL-1 beta, IL-2, and IL-6, also increased. These data suggest that the Coptis chinensis Franch. rhizome extract activates MOLT-4 cell to Th1 cell, not type II helper T cell. Furthermore, the Coptis chinensis Franch. rhizome extract activates the Mitogen-activated protein kinase (MAPKs) signaling pathways.Conclusion: The results obtained from this study suggest that the Coptis chinensis Franch. rhizome extract should be used as an immune enhancer in anti-inflammatory medicine, adjuvant materials, and as a supplement to treat weakened immune system. (C) 2016 Published by Elsevier Ireland Ltd.
Anthocyanin is a powerful natural antioxidant. Purple corn husk is rich in anthocyanin. In this paper the antioxidative effect of anthocyanin-rich purple corn husk extract (PCHE) in mayonnaise during storage was studied. The antioxidative effect of the mayonnaise containing PCHE was evaluated by measuring peroxide values, p-anisidine values, total oxidation values, acid values, and iodine values at time intervals for 10weeks. The antioxidative effect of the mayonnaise containing PCHE was higher than that of mayonnaise with chemical antioxidants BHT and EDTA as positive control. The mayonnaise containing 0.4g/kg PCHE showed the strongest antioxidative performance during storage. This study suggests that PCHE could be used as natural antioxidant in high fat food and as a substitute to chemical antioxidant with its purplish colour marking its difference from ordinary mayonnaise. Such colour difference will tell consumers that their food contains natural antioxidants.
Phytic acid in grains affects the bioavailability of minerals and nutrients in monogastric animals. Physicochemical properties of Sang-gol, a low-phytate rice cultivar developed to decrease anti-nutrient effect of phytic acid, were compared with those of its parent rice cultivar Il-pum. The amylose content of Sang-gol was lower, but its crude protein content was not significantly different compare with Il-pum. Texture profiles of cooked rice, except for hardness, adhesiveness and chewiness, did not show any differences. The hardness, adhesiveness and chewiness of the Sang-gol were lower than that of Il-pum. Pasting characteristics, peak viscosity, trough, and final viscosity of Sanggol were higher, and breakdown was lower, as compared to Il-pum. Setback of Il-pum was significantly (p <0.01) lower than Ilpum. The pasting temperatures of Sang-gol and Il-pum were very similar (68.1 and 68.0°C), respectively.
Among the extracts from 420 kinds of herbs, Prunus salicina, showing the highest glucosyltransferase inhibition activity, was purified and designated GTI-0163. Structural determination of GTI-0163 revealed it to be an oleic acid-based unsaturated fatty acid. GTI-0163 was an uncompetitive inhibitor of GTase. Among the unsaturated fatty acids, oleic acid showed a significantly higher GTase inhibitory activity than the saturated fatty acids or the ester form of oleic acid. These results strongly suggested that both the number of double bonds and the existence of free carboxyl groups of fatty acids play an important role in GTase inhibitory activity.
The major proteins in honey have different molecular weights depending upon the honeybee species. To confirm the origin of major honey proteins, honey protein produced by Apis cerana or Apis mellifera were purified and analyzed by MALDI-TOF. Two major proteins were identified as a major royal jelly protein 1. Although two major proteins shared primary structure, they showed different molecular weights of 56 and 59 kDa, respectively. To discriminate the honeybee species producing honey using SDS–PAGE, artificial marker proteins, 56 and 59 kDa, were produced from Escherichia coli. Two artificial marker proteins were co-electrophoresed with honey samples and the difference in molecular weight was readily distinguished by SDS–PAGE. Therefore, the measurement of major proteins in honey is a useful method to discriminate the honey that produced from different honeybee species.
Ginseng treated with several treatment conditions of various acids to search hydrolysates on the basis of increased biological activity and modified structure. In the result of acid treatment, the conversion rate of ginsenoside Rg3, Rk1 and Rg5 was highest when ginseng treated with citric acid. After added citric acid to ginseng extract, boiled at 100 degrees C for I hour and add enzyme, which is examined change by time. It compared with group which did not treated acid. Two groups became difference according to enzyme but the generation rate of ginsenoside Rg3, Rk1 and Rg5 did not show difference greatly. Also, the generation rate of ginsenoside Rg3, Rk1 and Rg5 by time passes did not show difference. The generation rate of ginsenoside Rg3, Rk1 and Rg5 increased when increased acid concentration, temperature and time. We did exclusion molding to shorten treatment time. In the result of ginseng treated with citric acid of various concentrations at various temperatures as time passes by extrusion molding, the generation rate of ginsenoside Rg3, Rk1 and Rg5 was highest when ginseng treated with 3% citric acid at 160 degrees C for 20 minutes. In addition, total saponin amount of ginseng treated with 3% citric acid at 160 degrees C for 20 minutes was about 11% higher than ginseng heated at 120 degrees C for 3 hours. These results indicated that our exclusion molding process more effective, compared to traditional red ginseng manufacturing process.
In this research, we screened for glucosyltransferase (GTase) inhibitors that effectively prevent the dental caries from 420 kinds of boiled water extracts of herbs and wild plants and searched for GTase inhibitory activities. Among them, 13 kinds of hot water extracts had high GTase inhibitory activities and especially, we Focused on Foeniculum vulgare which showed the highest inhibitory activity on GTase. The boiled water extract of E vulgare was stable at high temperature and showed as a mixed type of competitive and uncompetitive inhibition kinetic behavior. It did not have antibacterial effect on Streptococcus mutans and had inhibitory activity on GTase. Specially, in the clinical trial, the group treated by boiled water extract of F. vulgare showed more decrease of plague index at 4.8 point than untreated group. These results suggested that boiled water extract of E vulgare can effectively suppress the plague formation as it inhibits the GTase activity,.
Anthocyanin pigments are extracted from various plants and used for diverse purposes. The overall goal of this study was to develop high-anthocyanin corn to enhance the economic efficiency of anthocyanin production. We determined and compared the anthocyanin contents from the different parts of purple corn in various breeding lines. Our results revealed that purple corn produced the anthocyanin pigment throughout the plant, especially high in the husk and cob regions, although anthocyanin levels varied significantly among different plant parts. We analyzed the 295 selected lines from the 2006 breeding population, and it showed that anthocyanin levels of husks ranged from 17.3% to 18.9% of dry weight, roughly 10 times more than the standard current purple corn kernel content, 1.78%. LC-MS/MS analysis demonstrated that the main components of purple corn husk anthocyanin were cyanidin derivatives, and the most prevalent constituents were cyanidin-3-glucoside, cyanidin-3-succinylglucoside and pelargonidin-3-(6''-malonylglucoside). The results suggested that high-anthocyanin corn will boost the purple corn pigment production far more than its current level.