• Plant Breeding and Biotechnology
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• v.6 no.4
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• pp.413-425
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• 2018

A previous work developed and identified a new omega-5 gliadin deficient wheat line named O-free by crossing Keumkang and Olgeuru, which is nutritionally quite meaningful in that omega-5 gliadin is one of the known wheat allergens. To verify the characteristics of the O-free, we performed RNA sequencing (RNAseq) analysis of the O-free and the two parent lines (Keumkang and Olgeuru). The results of the similarity analysis with the ESTs for gliadins and glutenins showed that the O-free ESTs had no similarity with the omega-5 gliadin sequences but had similarity to other gliadins and glutenins. Furthermore, mapping results between the raw RNAseq data from the O-free and the omega-5 gliadin sequence showed a clear deletion of the N-terminal sequences which are an important signature of omega-5 gliadin. We also designed specific PCR primers that could identify omega-5 gliadin in the genomic DNA. The results showed that no omega-5 gliadin fragments were detected in the O-free. According to these results, we confirmed that the deficiency of omega-5 gliadin in the O-free is not caused by post-transcriptional or post-translational regulations such as epigenetic phenomena but by a simple deletion in the chromosome. Furthermore, we showed that the low-molecular weight glutenin subunit (LMW-GS) gene in the O-free had a single nucleotide polymorphism (SNP) causing a premature stop codon, resulting in a truncated polypeptide. We expect that the O-free line may serve as an excellent source of wheat that could prevail in the hypo-allergen wheat market, which has recently gained interest world-wide.

• Natural Product Sciences
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• v.28 no.2
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• pp.45-52
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• 2022

The identification of Plasmodium falciparum enoyl acyl-carrier protein reductase (pfENR) is considered as a potential biological target against malaria. Trema orientalis is considered a rich source of phytochemicals useful in malaria treatment. This study evaluated the in-vitro inhibitory activity of the extract and isolated compounds of T. orientalis leaf; the isolated compounds and the analogues of the most active compound were subjected to in-silico molecular docking studies on pfENR. The methanolic extract of T. orientalis was subjected to repeated chromatographic separation which led to the isolation of some compounds. The isolated compounds from the plant were examined for their antimalarial activity using β-hematin inhibition assay. Virtual screening via molecular docking and ADMET studies were conducted to gain insight into the mechanism of binding of ligand and to identify effective pfENR inhibitors. The isolated compounds and the analogues of the most active isolates were gotten from PubChem library for use in docking study. Hexacosanol and β-sitosterol showed inhibition of the β-hematin formation. The docking results showed that hexacosanol, β-sitosterol and the analogues of β-sitosterol displayed binding energy ranging between -6.1 kcal/mol and -11.6 kcal/mol. Sitosterol glucoside has the highest docking score. Some of the ligands showed more binding affinity than known bioactive compounds used as reference. Analogues of β-sitosterol has been shown to be potential inhibitors of pfENR, therefore, the findings from this study suggest that sitosterol glucoside and ergosterol peroxide could act as antimalarial agents after further lead optimisation investigations.

• Journal of The Korean Society of Grassland and Forage Science
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• v.43 no.2
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• pp.109-115
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• 2023

Ammonium (NH₄⁺) serves as a nitrogen source, but its elevated levels can hinder plant growth and production. Excess NH₄⁺ with α-ketoglutarate is assimilated into glutamate, a precursor of proline and glutathione (GSH). This study aimed to investigate the effects of excessive NH₄⁺ on the regulation of proline and GSH synthesis. Detached leaves from oilseed rape (Brassica napus L.) were fed with 0, 50, 100, 500, and 1000 mM NH₄Cl for 16 h. As the NH₄⁺ concentrations increased, the leaves exhibited progressive wilting and yellowing. Furthermore, total carotenoid and chlorophyll concentrations declined in response to all NH₄⁺ treatments, with the lowest levels observed in 1000 mM NH₄⁺ treatment. Hydrogen peroxide (H₂O₂) concentration showed a minor increase at low NH₄⁺ concentration (50 and 100 mM) treatments but a significant increase at high NH₄⁺ (500 and 1000 mM), which was consistent with the localization of H₂O₂. Amino acid concentrations increased with increasing in NH₄⁺ concentration, while the protein concentration displayed the opposite trend. Proline and cysteine concentrations exhibited a gradual increase in response to increasing NH₄⁺ concentrations. However, GSH concentrations rose only in the 50 mM NH₄⁺ treatment and decreased in the 500 and 1000 mM NH₄⁺ treatments. These results indicate that excessive NH₄⁺ is primarily assimilated into proline, while GSH synthesis is adversely affected.

• Journal of the Korean Society of Food Science and Nutrition
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• v.37 no.9
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• pp.1154-1161
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• 2008

The purpose of this study is to determine the possibility of using Sophorae fructus as natural health food source. To accomplish this purpose, the contents of general and antioxidative nutrients of Sophora fructus were measured. The contents of carbohydrate, crude protein, crude lipid and ash are 75.9%, 16.4%, 2.41%, and 5.2%, respectively, while the calories of Sophora fructus was 337.3 kcal. Total dietary fiber was 15.07% of total carbohydrates. The percentages of water soluble dietary fiber to insoluble dietary fiber were 1.09% and 10.36%, respectively. The protein was composed of a total of 18 different kinds of amino acids. The contents of essential and non-essential amino acids were 2,310.91 mg and 5,218.52 mg. The K was the largest mineral followed by Ca, P and Mg, which means Sophorae fructus is alkali material. The contents of saturated fatty acids, monounsaturated fatty acids and polyunsaturated fatty acids were 24.94%, 32.40%, and 32.86%, respectively. Therefore, the amount of the total unsaturated fatty acid was higher than that of any other plant. The content of vitamin C in Sophorae fructus was higher than that of any other plant, which suggests that it could increase blood elasticity. The content of rutin, which is responsible for capillary vessel permeability, was 1.78%. The contents of water soluble antioxidative materials in 1 mL of water-extracted Sophorae fructus were $4.95\;{\mu}g$ which is comparable to 1,560.96 mmol of vitamin C in antioxidant effect. The general nutrients and other antioxidatant bioactive materials in Sophorae fructus were also potential materials for good health food. It is expected that a follow up study on Sophorae fructus through developing processed food and evaluation of their functional properties would provide useful information as a source of medicinal foods.

• The Korean Journal of Food And Nutrition
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• v.28 no.4
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• pp.551-557
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• 2015

The genus Rosa (Rosaceae) is an abundant source of phenolics and is traditionally used as a food supplement and as herbal medicine. Various plant phenolics are known to have anticancer, antioxidant, and anti-inflammatory properties. In this study, we investigated the anti-inflammatory effects of rose methanolic extracts (RMEs) from four different rose cultivars (Macarena, Onnuri, Oklahoma, and Colorado) in lipopolysaccharide (LPS)-activated RAW 264.7 cells. Our results demonstrated that pretreatment of REMs ($500{\mu}g/mL$) significantly reduced NO production by suppressing iNOS protein expression in LPS-stimulated cells. Anti-inflammatory effects by RMEs were observed in the following order: Oklahoma > Colorado > Onnuri > Macarena. Consistent with this finding, RMEs inhibited the translocation of $NF-{\kappa}B$ from the cytosol to the nucleus via the suppression of $I{\kappa}B{\alpha}$ phosphorylation and also inhibited LPS-stimulated $NF-{\kappa}B$ transcriptional activity. These findings suggest that RMEs exert anti-inflammatory actions and help to elucidate the mechanisms underlying the potential therapeutic values of RMEs. Therefore, RMEs could be regarded as a potential source of natural anti-inflammatory agents.

• Journal of Life Science
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• v.31 no.11
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• pp.980-986
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• 2021

Natural products have always been an attractive source in terms of novel anti-metastatic compounds which can hinder MMP expression and activity. Corydalis heterocarpa is a salt marsh plant found in the seashores throughout Korea. Its yellow flowers and spikes have been an ingredient in folk medicine to treat spasm and contractions. The present study assessed the potential of different solvent-based fractions from the crude extract of Corydalis heterocarpa (CHE), a halophyte with reported bioactivities, to suppress the PMA-induced MMP expression in human fibrosarcoma HT-1080 cells. The solvent fractions which were named after the solvent used for fractionation (n-hexane, 85% aqueous (aq.) methanol (MeOH), n-butanol (BuOH), and H₂O were shown to inhibit the both elevated mRNA and protein expression levels of MMP-2 and MMP-9 and simultaneously relieved the suppression on the expression of the endogenous MMP inhibitors TIMP-1 and TIMP-2. Results indicated that the CHE fractions might intervene with the PMA-induced activation of the MAPK signaling which is the upstream activator of MMP overexpression. Among tested samples, 85% aq. MeOH and n-hexane fractions of CHE was determined to be the most active and future studies to isolate the bioactive substances responsible for the regulation of the MMP expression are, therefore, urged. In conclusion, C. heterocarpa was shown to be a potential source of anti-metastatic compounds and n-Hexane and MeOH fractions might yield lead molecules to develop novel MMP inhibitors.

• Asian-Australasian Journal of Animal Sciences
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• v.14 no.6
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• pp.880-884
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• 2001

Rumen microbiology research has undergone several evolutionary steps: the isolation and nutritional characterization of readily cultivated microbes; followed by the cloning and sequence analysis of individual genes relevant to key digestive processes; through to the use of small subunit ribosomal RNA (SSU rRNA) sequences for a cultivation-independent examination of microbial diversity. Our knowledge of rumen microbiology has expanded as a result, but the translation of this information into productive alterations of ruminal function has been rather limited. For instance, the cloning and characterization of cellulase genes in Escherichia coli has yielded some valuable information about this complex enzyme system in ruminal bacteria. SSU rRNA analyses have also confirmed that a considerable amount of the microbial diversity in the rumen is not represented in existing culture collections. However, we still have little idea of whether the key, and potentially rate-limiting, gene products and (or) microbial interactions have been identified. Technologies allowing high throughput nucleotide and protein sequence analysis have led to the emergence of two new fields of investigation, genomics and proteomics. Both disciplines can be further subdivided into functional and comparative lines of investigation. The massive accumulation of microbial DNA and protein sequence data, including complete genome sequences, is revolutionizing the way we examine microbial physiology and diversity. We describe here some examples of our use of genomics- and proteomics-based methods, to analyze the cellulase system of Ruminococcus flavefaciens FD-1 and explore the genome of Ruminococcus albus 8. At Illinois, we are using bacterial artificial chromosome (BAC) vectors to create libraries containing large (>75 kbases), contiguous segments of DNA from R. flavefaciens FD-1. Considering that every bacterium is not a candidate for whole genome sequencing, BAC libraries offer an attractive, alternative method to perform physical and functional analyses of a bacterium's genome. Our first plan is to use these BAC clones to determine whether or not cellulases and accessory genes in R. flavefaciens exist in clusters of orthologous genes (COGs). Proteomics is also being used to complement the BAC library/DNA sequencing approach. Proteins differentially expressed in response to carbon source are being identified by 2-D SDS-PAGE, followed by in-gel-digests and peptide mass mapping by MALDI-TOF Mass Spectrometry, as well as peptide sequencing by Edman degradation. At Ohio State, we have used a combination of functional proteomics, mutational analysis and differential display RT-PCR to obtain evidence suggesting that in addition to a cellulosome-like mechanism, R. albus 8 possesses other mechanisms for adhesion to plant surfaces. Genome walking on either side of these differentially expressed transcripts has also resulted in two interesting observations: i) a relatively large number of genes with no matches in the current databases and; ii) the identification of genes with a high level of sequence identity to those identified, until now, in the archaebacteria. Genomics and proteomics will also accelerate our understanding of microbial interactions, and allow a greater degree of in situ analyses in the future. The challenge is to utilize genomics and proteomics to improve our fundamental understanding of microbial physiology, diversity and ecology, and overcome constraints to ruminal function.

• Journal of Nutrition and Health
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• v.38 no.5
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• pp.352-363
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• 2005

The previous extensive in vitro studies on the antioxidative activities of a number of Korean grains, vegetables, seaweeds and mushrooms, and the various combinations of these food source exhibited a wide range of antioxidative activities, and four food mixtures composed of 5 kinds of foods (5A, 5B, 5C and 5D) were designed from 16 selective foods showing. high antioxidant effect, in vitro, to find the good combinations for the meal planning. Mixture 5B or 5C contained very high levels of total flavonoid and polyphenol, and ethanol extract from 5A, 5B or 5C showed very strong inhibitory effects against in vitro $Fe^{2+}-induced$ lipid peroxidation and ethanol extract from 5B or 5C showed remarkable DPPH radical scavenging effect and lipid peroxide-protein conjugation inhibition effect. And in vivo study was also carried out with two mixtures (5B, 5C). Powders (P5B, P5C) or ethanol extracts (E5B, E5C) of these mixtures were supplemented to Sprague-Dawley rats fed on high fat $(15\%)-high$ cholesterol $(1\%)$ semipurified diet for 5 weeks. The total antioxidant power in serum was significantly higher in P5B, P5C, E5B and E5C groups than in high fat control group, and $ascorbate-Fe^{2+}-induced$ TBARS was significantly lowered by E5B supplementation in rat liver. In liver tissue, Cu, Zn-SOD activity was significantly higher in P5B and E5B groups than in high fat control group, while catalase or GSH-peroxidase (GPx) activity was not changed by any supplementations. In kidney, Cu, Zn­SOD activity was significantly higher in P5B group than in high fat control group, while GPx activity was not changed by any supplementations. Taken together, mixture 5B and 5C showed very strong antioxidative effects both in vitro and in vivo. Therefore, the ingredient Korean foods of 5B and 5C could be recommended to take a lot together for prevention from age-related chronic diseases.

• Journal of the Korean Society of Food Science and Nutrition
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• v.38 no.9
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• pp.1139-1144
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• 2009

The purpose of this study is to determine the possibility of using Pueraria flos as natural health food source. To accomplish this purpose, the contents of general and antioxidative nutrients of Pueraria flos a were measured. The contents of carbohydrate, crude protein, crude lipid and ash were 69.2%, 19.9%, 2.2% and 8.9%, respectively, and calories was 340.4 kcal. Total dietary fiber was 85.1% of total carbohydrates. The percentages of water soluble dietary fiber to insoluble dietary fiber were 12.0% and 46.7%, respectively. The protein contained 18 different kinds of amino acids. The contents of essential and non-essential amino acids were 5.0 g and 6.7 g. The Ca was the largest mineral followed by K, Mg, and P, which means Pueraria flos is alkali material. The contents of saturated fatty acids, monounsaturated fatty acids and polyunsaturated fatty acids were 25.37%, 33.61% and 35.68%, respectively. Therefore, the amount of the total unsaturated fatty acid was higher than that of any other plant. The antioxidant activity of 70% ethanol extract and fractions of the Pueraria flos has been determined by the scavenging of the stable radical DPPH; the result showed that the ethyl acetate fraction was the most active, as the amount required for 50% reduction of DPPH after 30 mins ($RC_{50}$) was 109.9 ${\mu}g$, followed by 70% ethanol extract (217.3 ${\mu}g$), hexane fraction (134.5 ${\mu}g$), chloroform (116.7 ${\mu}g$), butanol faction (129.8 ${\mu}g$) and aqueous fraction (473.5 ${\mu}g$).

• Korean Journal of Food Science and Technology
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• v.37 no.6
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• pp.976-982
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• 2005

Abilities of various edible plants and natural antioxidants to protect brain against oxidative damages were evaluated using brain homogenate of perfused Sprague-Dawley rat. Oxidative damage, expressed as lipid peroxidation (LPO), indicating total quantity of malondialdehyde and 4-hydroxyalkenal, increased from 4.1 to 6.9nmol/mg protein by treatment of $2.5{\mu}M$ ferrous sulfate and 7.5mM hydrogen peroxide as source of reactive oxygen species (ROS) on brain homogenate for 10min at $37^{\circ}C$ Mallow(88%) in leafy vegetables, small potato (93%) in root vegetables, green red pepper (76%) in fruit vegetables, and avocado (96%) in fruits showed highest LPO inhibition capacities. Ability of mushrooms decreased in order of nameko, shiitake, pine mushroom, oyster mushroom, and new type pine mushroom. Among natural antioxidants tested, (+)catechin (91%), (-)epigallocatechin gallate (85%), (-)epicatechin gallate (83%), and kaempferol(83%) showed high LPO inhibition capacities.