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7,8,4'-Trihydroxyisoflavone, a Metabolized Product of Daidzein, Attenuates 6-Hydroxydopamine-Induced Neurotoxicity in SH-SY5Y Cells

  • Ko, Yong-Hyun;Kim, Seon-Kyung;Kwon, Seung-Hwan;Seo, Jee-Yeon;Lee, Bo-Ram;Kim, Young-Jung;Hur, Kwang-Hyun;Kim, Sun Yeou;Lee, Seok-Yong;Jang, Choon-Gon
    • Biomolecules & Therapeutics
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    • v.27 no.4
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    • pp.363-372
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    • 2019
  • Daidzein isolated from soybean (Glycine max) has been widely studied for its antioxidant and anti-inflammatory activities. However, the protective effects of 7,8,4'-trihydroxyisoflavone (THIF), a major metabolite of daidzein, on 6-hydroxydopamine (OHDA)-induced neurotoxicity are not well understood. In the current study, 7,8,4'-THIF significantly inhibited neuronal cell death and lactate dehydrogenase (LDH) release induced by 6-OHDA in SH-SY5Y cells, which were used as an in vitro model of Parkinson's disease (PD). Moreover, pretreatment with 7,8,4'-THIF significantly increased the levels of superoxide dismutase (SOD), catalase (CAT), and glutathione (GSH) and decreased malondialdehyde (MDA) activity in 6-OHDA-induced SH-SY5Y cells. In addition, 7,8,4'-THIF significantly recovered 6-OHDA-induced cleaved caspase-3, cleaved caspase-9, cleaved poly-ADP-ribose polymerase (PARP), increased Bax, and decreased Bcl-2 levels. Additionally, 7,8,4'-THIF significantly restored the expression levels of phosphorylated c-Jun N-terminal kinase (JNK), p38 mitogen-activated protein kinase (MAPK), extracellular signal-regulated kinase 1/2 (ERK 1/2), phosphatidylinositol 3-kinases (PI3K)/Akt, and glycogen synthase kinase-3 beta ($GSK-3{\beta}$) in 6-OHDA-induced SH-SY5Y cells. Further, 7,8,4'-THIF significantly increased the reduced tyrosine hydroxylase (TH) level induced by 6-OHDA in SH-SY5Y cells. Collectively, these results suggest that 7,8,4'-THIF protects against 6-OHDA-induced neuronal cell death in cellular PD models. Also, these effects are mediated partly by inhibiting activation of the MAPK and PI3K/Akt/$GSK-3{\beta}$ pathways.

Effect of in ovo feeding of γ-aminobutyric acid combined with embryonic thermal manipulation on hatchability, growth, and hepatic gene expression in broilers

  • Chris Major, Ncho;Akshat, Goel;Vaishali, Gupta;Chae-Mi, Jeong;Yang-Ho, Choi
    • Animal Bioscience
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    • v.36 no.2
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    • pp.284-294
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    • 2023
  • Objective: This study investigated the effects of in ovo feeding of γ-aminobutyric acid (GABA) and embryonic thermal manipulation (ETM) on growth performance, organ indices, plasma biochemical parameters, hepatic antioxidant levels, and expression of lipid metabolism-related genes in broilers. Methods: Two hundred and fifty eggs were assigned to one of four treatments: control eggs incubated under standard conditions (CON); eggs that received an in ovo injection of 10% GABA on day 17.5 of incubation (G10); thermally manipulated eggs between days 10 and 18 of incubation at 39.6°C for 6 h daily (TM); and eggs that received both treatments during incubation (G10+TM). After 28 days of rearing, five birds per treatment were selected for blood and organ sampling. Results: No differences were found in hatchability or growth parameters among different treatment groups. Hepatic gene expression of catalase (CAT) and glutathione peroxidase 1 (GPx1) was upregulated (p = 0.046 and p = 0.006, respectively) in the G10+TM group, while that of nuclear factor erythroid 2-related factor 2 (NRF2) was upregulated (p = 0.039) in the G10 group. In addition, the relative gene expression of NADPH oxidase 1 (NOX1) was significantly lower (p = 0.007) in all treatment groups than that in the CON group. Hepatic fatty acid synthase (FAS) levels and average daily feed intake (ADFI) of last week showed a positive correlation (r = 0.50, p = 0.038). In contrast, the relative gene expression of the extracellular fatty acid-binding protein (EXFAB) and peroxisome proliferator-activated receptor-γ (PPAR-γ) were positively correlated (r = 0.48, p = 0.042 and r = 0.50, p = 0.031) with the overall ADFI of birds. Conclusion: Taken together, the results of this study suggest that the combination of in ovo feeding of GABA and ETM can enhance hepatic antioxidant function in broilers.

Protective effect of Lycium barbarum leaf extracts on atopic dermatitis: in vitro and in vivo studies

  • Han Sol Lee;Eun Young Bae;Sun Yung Ly
    • Nutrition Research and Practice
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    • v.17 no.5
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    • pp.855-869
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    • 2023
  • BACKGROUND/OBJECTIVES: Atopic dermatitis (AD) is a chronic disease with an increasing incidence globally; therefore, there is a growing demand for natural compounds effective in treating dermatitis. In this study, the protective effects of Lycium barbarum leaves with and without chlorophyll (LLE and LLE[Ch-]) on AD were investigated in animal models of AD and HaCaT cells. Further, we investigated whether LLE and LLE(Ch-) show any differences in physiological activity. MATERIALS/METHODS: AD was induced by 2,4-dinitrochlorobenzene (DNCB) for three weeks, while NC/Nga mice were fed LLE or LLE(Ch-) extracts for 7 weeks. Serum immunoglobulin E (IgE) and cytokine (tumor necrosis factor [TNF]-α, interleukin [IL]-6, and IL-4) concentrations and the degree of DNA fragmentation in lymphocytes were examined. A histopathological examination (haematoxylin & eosin staining and blue spots of toluidine) of the dorsal skin of mice was performed. To elucidate the mechanism of action, the expression of the thymus and activation-regulated chemokine (TARC) and macrophage-derived chemokine (MDC) were measured in HaCaT cells. RESULTS: Serum IgE and cytokines (TNF-α and IL-6) levels as well as DNA fragmentation of lymphocytes were significantly decreased in AD-induced mice treated with LLE or LLE(Ch-) compared to those of the control group. The epidermal thickness of the dorsal skin and mast cell infiltration in the LLE group significantly reduced compared to that in the control group. The LLE extracts showed no cytotoxicity up to 1,000 ㎍/mL in HaCaT cells. LLE or LLE(Ch-)-treated group showed a reduction of TARC and MDC in TNF-α-and IFN-γ-stimulated HaCaT cells. CONCLUSIONS: These results suggest that LLE potentially improves inflammation by reducing the expression of chemokines that inhibit T helper 2 cell migration. LLE(Ch-) showed similar effects to LLE on blood levels of IgE, TNF-α and IL-6 and protein expression in HaCat cells, but the ultimate effect of skin improvement was not statistically significant. Therefore, both LLE and LLE(Ch-) can be used as functional materials to alleviate AD, but LLE(Ch-) appears to require more research to improve inflammation.

The Effect of Antioxidative Change in Cardiac Muscle of Obesity Rat by Treadmill Exercise with Intensity and Time (운동강도와 지속시간에 따른 트레드밀 운동이 비만 쥐의 심장근 내 항산화에 미치는 영향)

  • Kim, Myung-Hee;Kim, Young-Eok;Yoon, Chang-Lyuk;Ryu, Ji-Won;Ahn, Jong-Mo
    • Journal of Oral Medicine and Pain
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    • v.38 no.1
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    • pp.35-51
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    • 2013
  • The aims of this study was to observed an effect of antioxidative in cardiac muscle of high fat diet induced obesity rat by treadmill exercise with intensity and time. Thirty-two Sprauge-Dawley rats which were divided into four group. Normal, Control(high fat diet induced obesity rat), Experimental I(high intensity intermittent exercise in high fat diet induced obesity rat), Experimental II(moderate intensity endurance exercise in high fat diet induced obesity rat). The results of this study were as follows: 1. In change of body weight, the outcome of each group significantly difference compared with control. Also, 1 to 3 weeks significantly different compared with pre valu experimental I and II(p<0.001). 2. In change of lipid profile, the outcome of each group significantly difference compared with control(p<0.001). Difference between experimental I and II is not significantly. 3. In change of antioxidative enzymes(SOD, CAT, GPx) in myocardium, there are significant difference between control and experimental II, and also between control and experimental I(p<0.001). 4. In change of antioxidative protein MCR-1, the outcome of each group significantly difference compared with control(p<0.01). Experimental II was most significantly difference than the other group(p<0.001). The above results suggest that treadmill exercise effectively reduced in fat. It would be considered that moderate intensity endurance exercise has an effects on improved antioxidative enzyme in cardiac muscle of high fat diet induced obesity rat.

Characterization of Plasmid pKJ36 from Bifidobacterium longum and Construction of an E. coli-Bifidobacterium Shuttle Vector

  • Park, Nyeong-Soo;Shin, Dong-Woo;Lee, Ke-Ho;Ji, Geun-Eog
    • Journal of Microbiology and Biotechnology
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    • v.10 no.3
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    • pp.312-320
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    • 2000
  • Abstract The full sequence of the plasmid pKJ36, which was derived from Bifidobacterium longum KJ, was determined and analyzed to construct shuttle vectors between E. coli and Bifidobacterium. The plasmid pKJ36 was composed of 3,625 base pairs with a 65.1% G+C content. The structural organization of pKJ36 was highly similar to that of pKJ50, and the three major ORFs on pKJ36 showed high amino acid sequence homologies with those of pKJ50. The putative proteins coded by these three ORFs were designated as RepB (32.0 kDa, pI=9.25), MembB (29.0 kDa, pI=12.25), and MobB (39.0 kDa, pI=IO.66), respectively. The amino acid sequence of RepB showed a 57% identity and 70% similarity with that of the RepA protein of pKJ50. Upstream of the repB gene, the so-called iteron sequence was directly repeated four-and-ahalf times and a conserved dnaA box was identified. An amino acid sequence comparison between the MobB and MobA of pKJ50 revealed a 48% identity and 61 % similarity. A conserved oriT sequence with an inverted repeat identical to that of pKJ50 was also found upstream of the mobB gene. A hydropathy analysis of MembB revealed four possible transmembrane regions. The expressions of the repB and membB genes were confirmed by RT-PCR. The in vitro translation reaction of pKJ36 showed protein bands with anticipated sizes with respect to each putative gene product. S 1 endonuclease treatment and Southern hybridization suggested that pKJ36 replicates by a rolling circle mechanism via a single-stranded DNA (ssDNA) intermediate. A shuttle vector between E. coli and Bifidobacterium sp. was constructed using the pKJ36, pBR322, and staphylococcal chloramphenicol acetyl transferase (CAT) gene. The successful transformation of the Bifidobacterium strains was shown by Southern hybridization and PCR. The transformation efficiency differed from strain to strain and, depending on the electroporation conditions, with a range between $1.2{\times}10^1-2.6{\times}10^2{\;}cfu/\mu\textrm{g}$ DNA.X> DNA.

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Effect of Electroacupuncture Analgesia on Changes of Vital Signs and Blood Chemical Values in Cats

  • Shin, Dong-Hoon;Lee, Jae-Yeon;Kim, Duck-Hwan;Park, Chang-Sik;Jeong, Seong-Mok;Son, Dong-Soo;Kim, Myung-Cheol
    • Journal of Veterinary Clinics
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    • v.25 no.3
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    • pp.170-175
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    • 2008
  • The present study was performed to investigate the anesthetic or analgesic effect of tiletamine-zolazepam (TZ) and electro acupuncture analgesia (EAA) in cats. Twelve healthy cats were randomly assigned to receive either TZ or EA. TZ group cats with weight of $3.65{\pm}0.48kg$ received 10.0 mg/kg of TZ intramuscularly. EA group cats with weight of $3.62{\pm}0.52kg$ received 5V, 30Hz and 60 minutes of EA. The acupoints used were Tian-ping (GV-5, +), Bai-hui (GV-20, -). Therefore, after and before experiment, some serum chemistry profiles (alkaline phospatase, aspartate aminotransferase, alanine aminotransferase, glucose and total protein) and change of vital signs (rectal temperature, heart rate, respiratory rate) were examined. All cats were examined pre, and 5, 25, 65 and 105 minutes after administration of TZ or operation of EA. The cats in EA group showed a smaller change in rectal temperature, heart rate and respiratory rate than in the TZ group (p<0.05). In both groups, total protein concentration was constant throughout the period of anesthesia, and the serum glucose increased gradually throughout the period of anesthesia. However, the cats in EA group showed a smaller change in alkaline phospatase, aspartate aminotransferase and alanine aminotransferase within the limit of safety than in the TZ group (p<0.05). While coming to induction, the TZ group took a mean $2.4{\pm}0.7$ minutes to achieve sternal recumbency, compared with $10.5{\pm}2.1$ minutes by the EA group, and $3.2{\pm}0.6$ minutes to achieve lateral recumbency, compared with $18.8{\pm}1.9$ minutes by the EA group (p<0.05). When recovering from anesthesia, the TZ group took $164.3{\pm}17.9$ minutes to achieve sternal position time, compared with $67.7{\pm}4.6$ minutes by the EA group, and $202.0{\pm}15.7$ minutes to stand, compared with $73.0{\pm}6.1$ minutes for the EA group (p<0.05). In this study, the cats anesthetized with EA showed a more rapid recovery rather than the cats under TZ anesthesia. Also, there do not appear to be any negative physiologic effects associated with acupuncture-induced surgical analgesia. So, it was considered that EAA may be used effectively in shock, debilitated cats, as compared to TZ.

Cloning and Overexpression of a Paenibacillus ${\beta}-Glucanase$ in Pichia pastoris: Purification and Characterization of the Recombinant Enzyme

  • Yang, Peilong;Shi, Pengjun;Wang, Yaru;Bai, Yingguo;Meng, Kun;Luo, Huiying;Yuan, Tiezheng;Yao, Bin
    • Journal of Microbiology and Biotechnology
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    • v.17 no.1
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    • pp.58-66
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    • 2007
  • Isolation, expression, and characterization of a novel $endo-{\beta}-1,3(4)-D-glucanase$ with high specific activity and homology to Bacillus lichenases is described. One clone was screened from a genomic library of Paenibacillus sp. F-40, using lichenan-containing plates. The nucleotide sequence of the clone contains an ORF consisting of 717 nucleotides, encoding a ${\beta}-glucanase$ protein of 238 amino acids and 26 residues of a putative signal peptide at its N-terminus. The amino acid sequence showed the highest similarity of 87% to other ${\beta}-1,3-1,4-glucanases$ of Bacillus. The gene fragment Bg1 containing the mature glucanase protein was expressed in Pichia pastoris at high expression level in a 3-1 high-cell-density fermenter. The purified recombinant enzyme Bg1 showed activity against barley ${\beta}-glucan$, lichenan, and laminarin. The gene encodes an $endo-{\beta}-1,3(4)-D-glucanase$ (E. C. 3.2.1.6). When lichenan was used as substrate, the optimal pH was 6.5, and the optimal temperature was $60^{\circ}C$. The $K_m,\;V_{max},\;and\;k_{cat}$ values for lichenan are 2.96mg/ml, $6,951{\mu}mol/min{\cdot}mg,\;and\;3,131s^{-1}$, respectively. For barley ${\beta}-glucan$ the values are 3.73mg/ml, $8,939{\mu}mol/min{\cdot}mg,\;and\;4,026s^{-1}$, respectively. The recombinant Bg1 had resistance to pepsin and trypsin. Other features of recombinant Bg1 including temperature and pH stability, and sensitivity to some metal ions and chemical reagents were also characterized.

Antioxidant capacity in seedling of colored-grain wheat under water deficit condition

  • Kim, Dae Yeon;Hong, Min Jeong;Jung, Woo Joo;Seo, Yong Weon
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.140-140
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    • 2017
  • Nutritious and functional foods from crop have received great attention in recent years. Colored-grain wheat contains high phenolic compound and a large number of flavonoid. The anthocyanin and polyphenolic synthesis and accumulation is generally stimulated in response to biotic or abiotic stresses. Here, we analyzed genome wide transcripts in seedling of colored-grain wheat response to ABA and PEG treatment. About 900 and 1500 transcripts (p-value < 0.05) from ABA and PEG treatment were aligned to IWGSC1+popseq DB which is composed of over 110,000 transcripts including 100,934 coding genes. NR protein sequences of Poaceae from NCBI and protein sequence of transcription factors originated from 83 species in plant transcription factor database v3.0 were used for annotation of putative transcripts. Gene ontology analysis were conducted and KEGG mapping was performed to show expression pattern of biosynthesis genes related in flavonoid, isoflavonoid, flavons and anthocyanin biopathway. DroughtDB (http://pgsb.helmholtz-muenchen.de/droughtdb/) was used for detection of DEGs to explain that physiological and molecular drought avoidance by drought tolerance mechanisms. Drought response pathway, such as ABA signaling, water and ion channels, detoxification signaling, enzymes of osmolyte biosynthesis, phospholipid metabolism, signal transduction, and transcription factors related DEGs were selected to explain response mechanism under water deficit condition. Anthocyanin, phenol compound, and DPPH radical scavenging activity were measured and antioxidant activity enzyme assays were conducted to show biochemical adaptation under water deficit condition. Several MYB and bHLH transcription factors were up-regulated in both ABA and PEG treated condition, which means highly expressed MYB and bHLH transcription factors enhanced the expression of genes related in the biosynthesis pathways of flavonoids, such as anthocyanin and dihydroflavonols in colored wheat seedlings. Subsequently, the accumulation of total anthocyanin and phenol contents were observed in colored wheat seedlings, and antioxidant capacity was promoted by upregulation of genes involved in maintaining redox state and activation of antioxidant scavengers, such as CAT, APX, POD, and SOD in colored wheat seedlings under water deficit condition. This work may provide valuable and basic information for further investigation of the molecular responses of colored-grain wheat to water deficit stress and for further gene-based studies.

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Changes of antioxidant enzyme activities subjected to water stress in soybean leaves (대두(大豆)에서 분석(水分)스트레스에 의(依)한 항산화효소(抗酸化酵素)의 활성도(活性度) 변화(變化))

  • Kim, Tae-Sung;Kang, Sang-Jae;Park, Woo-Churl
    • Current Research on Agriculture and Life Sciences
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    • v.16
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    • pp.24-30
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    • 1998
  • This experiment was carried out to elucidate and study about plant defense mechanism subjected to water stress(drought, flooding). We measured water content, total soluble protein content as stress marker and superoxide dismutase(SOD), catalase (CAT) as antioxidant enzymes subjected to water stress(drought, flooding) and recovery in soybean leaves. The results obtained were as follows; Two soybean lines(keonolkong, euhakong)leaves exposed to water stress (drought, flooding) showed premature senescence as evidence by the decrease in water content, and total soluble protein content, but those of soybean leaves subject to water stress recovery for 3 days were recovered. Visual damage was much worse at drought stress than flooding stress and was worse keunolkong than enhakong. The activity of superoxide dismutase, catalase subjected to water stress(drought, flooding)was on the decrease, but degree of decrease was different from a sort of soybean lines, drought and flooding stress.

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Antioxidant Activity of Novel Casein-Derived Peptides with Microbial Proteases as Characterized via Keap1-Nrf2 Pathway in HepG2 Cells

  • Zhao, Xiao;Cui, Ya-Juan;Bai, Sha-Sha;Yang, Zhi-Jie;Cai, Miao;Megrous, Sarah;Aziz, Tariq;Sarwar, Abid;Li, Dong;Yang, Zhen-Nai
    • Journal of Microbiology and Biotechnology
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    • v.31 no.8
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    • pp.1163-1174
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    • 2021
  • Casein-derived antioxidant peptides by using microbial proteases have gained increasing attention. Combination of two microbial proteases, Protin SD-NY10 and Protease A "Amano" 2SD, was employed to hydrolyze casein to obtain potential antioxidant peptides that were identified by LC-MS/MS, chemically synthesized and characterized in a oxidatively damaged HepG2 cell model. Four peptides, YQLD, FSDIPNPIGSEN, FSDIPNPIGSE, YFYP were found to possess high 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging ability. Evaluation with HepG2 cells showed that the 4 peptides at low concentrations (< 1.0 mg/ml) protected the cells against oxidative damage. The 4 peptides exhibited different levels of antioxidant activity by stimulating mRNA and protein expression of the antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px), as well as nuclear factor erythroid-2-related factor 2 (Nrf2), but decreasing the mRNA expression of Kelch-like ECH-associated protein 1 (Keap1). Furthermore, these peptides decreased production of reactive oxygen species (ROS) and malondialdehyde (MDA), but increased glutathione (GSH) production in HepG2 cells. Therefore, the 4 casein-derived peptides obtained by using microbial proteases exhibited different antioxidant activity by activating the Keap1-Nrf2 signaling pathway, and they could serve as potential antioxidant agents in functional foods or pharmaceutic preparation.