Objective: p66Shc, a 66 kDa protein isoform encoded by the proto-oncogene SHC, is an essential intracellular redox homeostasis regulatory enzyme that is involved in the regulation of cellular oxidative stress, apoptosis induction and the occurrence of multiple age-related diseases. This study investigated the expression profile and functional characteristics of p66Shc during preimplantation embryo development in sheep. Methods: The expression pattern of p66Shc during preimplantation embryo development in sheep at the mRNA and protein levels were studied by quantitative real-time polymerase chain reaction (RT-qPCR) and immunofluorescence staining. The effect of p66Shc knockdown on the developmental potential were evaluated by cleavage rate, morula rate and blastocyst rate. The effect of p66Shc deficiency on reactive oxygen species (ROS) production, DNA oxidative damage and the expression of antioxidant enzymes (e.g., catalase and manganese superoxide dismutase [MnSOD]) were also investigated by immunofluorescence staining. Results: Our results showed that p66Shc mRNA and protein were expressed in all stages of sheep early embryos and that p66Shc mRNA was significantly downregulated in the 4-to 8-cell stage (p<0.05) and significantly upregulated in the morula and blastocyst stages after embryonic genome activation (EGA) (p<0.05). Immunofluorescence staining showed that the p66Shc protein was mainly located in the peripheral region of the blastomere cytoplasm at different stages of preimplantation embryonic development. Notably, serine (Ser36)-phosphorylated p66Shc localized only in the cytoplasm during the 2- to 8-cell stage prior to EGA, while phosphorylated (Ser36) p66Shc localized not only in the cytoplasm but also predominantly in the nucleus after EGA. RNAi-mediated silencing of p66Shc via microinjection of p66Shc siRNA into sheep zygotes resulted in significant decreases in p66Shc mRNA and protein levels (p<0.05). Knockdown of p66Shc resulted in significant declines in the levels of intracellular ROS (p<0.05) and the DNA damage marker 8-hydroxy2'-deoxyguanosine (p<0.05), markedly increased MnSOD levels (p<0.05) and resulted in a tendency to develop to the morula stage. Conclusion: These results indicate that p66Shc is involved in the metabolic regulation of ROS production and DNA oxidative damage during sheep early embryonic development.
Lingmin Jiang;Hanna Choe;Yuxin Peng;Doeun Jeon;Donghyun Cho;Yue Jiang;Ju Huck Lee;Cha Young Kim;Jiyoung Lee
Journal of Microbiology and Biotechnology
/
v.33
no.10
/
pp.1292-1298
/
2023
PAMB 00755T, a bacterial strain, was isolated from Korean fir leaves. The strain exhibits yellow colonies and consists of Gram-negative, non-motile, short rods or ovoid-shaped cells. It displays optimal growth conditions at 20℃, 0% NaCl, and pH 6.0. Results of 16S rRNA gene-based phylogenetic analyses showed that strain PAMB 00755T was most closely related to Sphingomonas chungangi MAH-6T (97.7%) and Sphingomonas polyaromaticivorans B2-7T (97.4%), and ≤96.5% sequence similarity to other members of the genus Sphingomonas. The values of average nucleotide identity (79.9-81.3%), average amino acid identity (73.3-75.9%), and digital DNA-DNA hybridization (73.3-75.9%) were significantly lower than the threshold values for species boundaries; these overall genome-related indexes (OGRI) analyses indicated that the strain represents a novel species. Genomic analysis revealed that the strain has a 4.4-Mbp genome encoding 4,083 functional genes, while the DNA G+C content of the whole genome is 66.1%. The genome of strain PAMB 00755T showed a putative carotenoid biosynthetic cluster responsible for its antioxidant activity. The respiratory quinone was identified as ubiquinone 10 (Q-10), while the major fatty acids in the profile were identified as C18:1ω7c and/or C18:1ω6c (summed feature 8). The major polar lipids of strain PAMB 00755T were diphosphatidylglycerol, phosphatidylethanolamine, sphingoglycolipid, and phosphatidylcholine. Based on a comprehensive analysis of genomic, phenotypic, and chemotaxonomic characteristics, we proposed the name Sphingomonas abietis sp. nov. for this novel species, with PAMB 00755T as the type strain (= KCTC 92781T = GDMCC 1.3779T).
Proceedings of the Plant Resources Society of Korea Conference
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2021.04a
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pp.37-37
/
2021
Calystegia soldanella L. is a perennial herbaceous halophyte belonging to the convolvulaceae family, which mainly grows in coastal sand dunes in Korea. Shoots and rhizomes are edible, and roots called 'Hyoseon Chogeun' are known to have medicinal effects such as antipyretic, sterilization, and diuretic. In addition, physiological activities of antioxidant, anti-inflammatory, antiviral, antifungal and PTP-1B (protein tyrosine phosphate-1B) inhibition have been reported. In this study, in vitro induction cell lines of C. soldanella L. collected from the coastal sand dunes in Jeju island was redifferentiated into adventitious roots that can be used as medicinal resources. Also the biomass of mass-proliferated adventitious roots using a bioreactor were evaluated. Plants of C. soldanella L. were collected from the crevice of the seashore in the coastal area of Taeheung 2-ri, Namwon-eup, Seogwipo-si. Then, it was separated into leaves, stems, rhizomes, and roots, and surface sterilized with 70% ethyl alcohol and 2% NaOCl (sodium hypochlorite). After washing with sterilized water, each organ section was cultured in Hormone-free MS medium (Murashige & Skoog Medium). As a result, the induction response rates were evaluated at 85% and 55%, respectively, in terms of callus formation and shoot generation in the rhizome segment. In the case of the adventitious roots morphological characteristics induced by single-use treatment of auxin-based plant growth regulators IBA and NAA from redifferentiated shoots were compared. Most efficient adventitious root culture method as a rooting rate, number, length, and biomass proliferation in the bioreactor system was confirmed when treated by culturing in MS salts, Sucrose 30 g·L-1, and IBA 1mg·L-1 for 4 weeks. In this study, the medium composition and culture period were confirmed using a bioreactor system to mass-proliferate adventitious roots derived from C. soldanella L. in Jeju island. Also this adventitious root line developed a new medicinal material could increase value of the bio-industry ingredient through quantitative and qualitative screening of phyto-bioactive compounds.
This study was conducted to develop a preservation technology that can induce changes in physicochemical properties to effectively utilize of persimmon leaves. The application effects of steaming or freezing technique were investigated. Astringent persimmon leaves were steam-blanched (100℃, 30 sec) or frozen (-20℃, 15 d), followed by hot-air drying (50℃). The physicochemical properties of the extract obtained by hot-water leaching from the dried leaves were compared. The extract of leaves dried without pretreatment was used as a control. L* value was higher in steamed than in control and frozen. a* value was highest in the control. The browning index was higher in the frozen and lower in the steamed than in the control. Soluble solids were the highest in the steamed and the lowest in the frozen. Sucrose content was relatively high in the steamed, and the glucose and fructose contents were relatively high in the frozen. Total polyphenol content and DPPH radical scavenging activity were higher in steamed and lower in frozen than in control. Thus, it was confirmed that steam or freeze pretreatment after harvesting persimmon leaves affects the extraction yield, color, antioxidant capacity and component changes of dried persimmon leaves. Unlike steaming, freezing pretreatment showed the effect of promoting decomposition and browning reactions, and it is considered useful when such an effect is needed.
Germination is controlled by internal factors of seeds and external factors such as water, temperature and light. We investigated the relationship between germination characteristics of four waxy rice cultivars and patterns of water uptake, antioxidant enzymes and free soluble sugars during early imbibition. Seed viabilities by tetrazolium test of four different rice cultivars were higher than 95% and germination rates of the hulled rice seeds were on 95% average. However, germination rate of intact rice among four cultivars showed a big difference depending on temperature. Water uptake of hulled and intact rice seeds during imbibition reached a stationary phase at around 30% moisture content. Although rates of water uptake were faster in hulled rice and high temperature than intact rice and low temperature condition, difference of those among cultivars was greater under low temperature than high temperature. The time required for rice seeds to uptake 30% water was negatively correlated with percentage of germination, germination energy, germination speed and mean germination time. Guaiacol peroxidase activity at 24h of imbibition was correlated with germination energy and germination speed but not percentage of germination. Catalase activity, soluble protein and maltose concentration at 24h of imbibition were not correlated with characteristics of germination. These results suggest that a time required for rice seeds to uptake 30% of water significantly correlated with germination and guaiacol peroxidase activity during early imbibition plays an important role in initiation of germination.
Journal of the Korean Society of Food Science and Nutrition
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v.38
no.10
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pp.1406-1413
/
2009
In order to investigate the quality characteristics of candies prepared with Compositae species flower extracts, the samples of Chrysanthemum indicum Linne (CI), C. boreale Makino (CB), and C. zawadskii var. latilobum (Maxim.) Kitam (CZ) were extracted with hot water and concentrated, and then added with various concentrations of 5$\sim$20% for candy preparation. L-value of candy color was decreased, but a- and b-value were increased with increasing amount of added extracts. pH was decreased, but total acidity was increased with increasing amount of added extracts. Moisture content of CI, CB, and CZ candies were 2.61$\sim$4.11, 2.46$\sim$4.57, and 2.27$\sim$4.43%, respectively. Hardness of candies were decreased with increasing amount of added extracts. CZ candy of 20% added extract showed the highest total polyphenol, flavonoid content, electron donating ability and anol, flavonaid content at 2.71 mg/g, 0.86 mg/g, 76.24%, and 3.17 mg AA eq/g, respectively. In sensory evaluation, the CI and CZ candies of 10% added extract obtained higher sensory scores.
Jeong, Yun Sook;Hwang, Kyung-A;Kim, Ga Ram;Song, Jin;Noh, Geon Min;Hwang, In Guk
The Korean Journal of Food And Nutrition
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v.28
no.5
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pp.745-751
/
2015
This study was performed to investigate the quality characteristics such as pH, total acidity, browning index, 5-hydroxymethylfurfural (5-HMF), fructose, S-allyl-L-cysteine (SAC), total polyphenol, and DPPH radical scavenging activity of aged garlic at different aging temperatures and periods. Aging temperature and period had a significant (p<0.05) effect on the quality characteristics of garlic. The pH in aged garlic significantly (p<0.05) decreased with increasing aging temperatures and periods, while the total acidity, browning index, and 5-HMF levels increased. The 5-HMF levels in garlic aged at 60 and $70^{\circ}C$ increased from 0.08~3.30 and from 0.05~106.07 mg/100 g, respectively. The fructose content in garlic aged at $60^{\circ}C$ gradually increased from 0.77 to 14.57%, while that of garlic aged at $70^{\circ}C$ increased from 1.07 to 19.75% until day 30, after which it decreased. The SAC level in raw garlic was 47.09 mg/100 g. The SAC contents of aged garlic differed significantly according to aging temperature and period. The SAC levels in garlic aged at $60^{\circ}C$ and $70^{\circ}C$ were in the range of 15.28~100.5 and 12.41~85.55 mg/100 g, respectively. The total polyphenol contents of garlic aged at $60^{\circ}C$ and $70^{\circ}C$ were 6.01 and 6.67 fold higher, respectively, than those in raw garlic. The DPPH radical scavenging activity of aged garlic also showed a tendency to increase during aging.
Brown color characteristics and antioxidizing activities were investigated for Doenjang under different processing conditions. Doenjang A was prepared directly from Meju and saline solution whereas Doenjang B was Prepared after separating soy sauce by soaking for 45 days. Both Doenjangs were aged for up to 180 days. Antioxidizing activity was studied in relation to the brown color characteristics using fat-soluble extract and water-soluble extract of Doenjang. The intensity of brown color was higher in the water-soluble Doenjang extract than the fat-soluble Doenjang extract. In the UV-VIS scanning spectra, water-soluble Doenjang extracts showed significant changes as the aging proceeded, but fat-soluble Doenjang extract did not. Antioxidizing activity of fat-soluble Doenjang extract increased as the aging period extended; however, no significant difference was detected in the water-soluble extract. Overall, Doenjang A showed higher contents of amino acids, reducing sugar, brown color, and antioxidizing activity, and the antioxidizing ability was higher in water-soluble Doenjang extract rather than in the fat-soluble Doenjang extract.
Kim, Da-Mi;Kim, Kyoung-Hee;Yun, Young-Sik;Kim, Jae-Hun;Lee, Ju-Woon;Yook, Hong-Sun
Journal of the Korean Society of Food Science and Nutrition
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v.40
no.10
/
pp.1460-1468
/
2011
This study investigated the quality changes and characteristics (0, 3, 6, 9%) of pound cake made with flour that included gamma irradiated (50 kGy) hot water extracts of Undaria pinnatifida sporophyll (WEUS). The pH of pound cakes decreased with increasing powder concentration, and gamma-irradiated pound cakes had lower pH than non-irradiated pound cakes at the same powder concentrations. The height, volume, specific loaf volume, and baking loss showed no significant differences between control and experimental groups. With increasing powder concentration, the L value of the crust and crumbs decreased, but the a value increased. The b value showed different tendencies between crust and crumb. The crust value was reduced with higher content of WEUS, but the crumb value increased. Gamma-irradiated pound cakes were also less hard than non-irradiated pound cakes. On the other hand, adhesiveness and springiness decreased with increasing powder concentration, but were not significantly different from the control. Also, gumminess and chewiness decreased but not significantly so. The hardness after several days of storage (5, 10, and 15 days) was higher than the control, and the springiness and cohesiveness were significantly reduced with increasing concentration compared to the control. The retrogradation increased in the control group, but it did not in the experimental groups. Results of radical scavenging activity using DPPH indicated that the gamma-irradiated group was higher than the non-irradiated group and it was also higher with higher concentrations of powder. In a sensory evaluation, when compared to the control, pound cake with 3% WEUS was superior in taste, flavor, and overall preference. Therefore, it was found that pound cake with 3% WEUS powder with gamma irradiation of 50 kGy added could improve the yield, taste, and antioxidant activity of pound cake.
This study was carried out to compare the growth characteristics, biological activities, β-glucan contents, sugar contents, and amino acid contents of 14 strains of Ganoderma spp. Among the 14 strains of Ganoderma spp., KMCC02960 (G. meredithae) and KMCC02932 (G. tropicum) showed excellent growth characteristics such as those with respect to the size and yield of fruiting bodies. The highest 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity was observed in KMCC02932 (G. tropicum). The nitrite scavenging activities of KMCC02824 (G. lucidum) and KMCC02852 (G. neo-japonicum) were higher than those of the other strains. The total polyphenol contents of the extracts from KMCC02824 (G. lucidum) and KMCC02852 (G. neo-japonicum) were higher than those of the other strains. KMCC03018 (G. lingzhi) showed the highest β-glucan content of 33.4%. In an analysis of the 4 types of monosaccharides, 2 types of disaccharides, and 4 types of sugar alcohols, only KMCC02996 (G. weberianum) and KMCC03018 (G. lingzhi) were commonly detected out of the 14 strains of Ganoderma spp. Eighteen amino acids, including eight essential amino acids, were identified: the highest total amino acids and total essential amino acids were found in KMCC02932 (G. tropicum), which had the highest levels of tyrosine and phenylalanine. Although the contents of amino acids differed by strain, cysteine, tyrosine, and phenylalanine were the most abundant amino acids in the analyzed extracts.
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