• Title/Summary/Keyword: Non-enzymatic

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Inhibition of MMP-2 and MMP-9 activities by solvent-partitioned Sargassum horneri extracts

  • Karadeniz, Fatih;Lee, Seul-Gi;Oh, Jung Hwan;Kim, Jung-Ae;Kong, Chang-Suk
    • Fisheries and Aquatic Sciences
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    • v.21 no.6
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    • pp.16.1-16.7
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    • 2018
  • Background: Matrix metalloproteinases (MMPs) are linked with several complications such as metastasis of cancer progression, oxidative stress, and hepatic fibrosis. Brown seaweeds are being extensively studied for their bioactive molecule content against cancer progression. In this context, Sargassum horneri was reported to possess various bioactivities including antiviral, antimicrobial, and anti-inflammatory partly due to its phenolic compound content. Methods: In this study, potential of S. horneri was evaluated through anti-MMP effect in HT1080 fibrosarcoma cells. S. horneri crude extract was fractionated with organic solvents, namely, water ($H_2O$), n-buthanol (n-BuOH), 85% aqueous methanol (85% aq. MeOH), and n-hexane. The non-toxicity of fraction samples (Sargassum horneri solvent-partitioned extracts (SHEs)) was confirmed by cell-viability assay. SHEs were tested for their ability to inhibit MMP enzymatic activity through gelatin digestion evaluation and cell migration assay. Expressions of MMP-2 and MMP-9 and tissue inhibitors of MMP (TIMPs) were evaluated by reverse transcription and Western blotting. Results: All fractions inhibited the enzymatic activities of MMP-2 and MMP-9 according to gelatin zymography. Except $H_2O$ fraction, fractions hindered the cell migration significantly. All tested fractions suppressed both mRNA and protein levels of MMP-2, MMP-9, TIMP-1, and TIMP-2. Conclusion: Overall, current results suggested that S. horneri has potential to be a good source for anti-MMP agents, and further investigations are underway for better understanding of the action mechanism and isolation and elucidation of the bioactive molecules.

7, 8, 4'-Trihydroxyisoflavone from Fermented Soybean Food and Its Biological Activity (전통 콩발효 식품 유래 7, 8, 4'-Trihydroxyisoflavone의 피부 생리활성 연구)

  • Park, Jun-Seong;Kim, Dong-Hyun;Shim, Jin-Sup;Kim, Ji-Seong;Choi, Kwon-Young;Kim, Byung-Gee;Kim, Duck-Hee;Kim, Han-Kon
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.36 no.3
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    • pp.221-226
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    • 2010
  • A rare isoflavone, 7, 8, 4'-trihydroxyisoflavone, was isolated from a 10-year-old fermented soybean food. 7, 8, 4'-trihydroxyisoflavone was isolated for the first time from a Korean fermented soybean food. Evaluation tests of biological activity showed significantly inhibition activity for free radical scavenging on both non-enzymatic (DPPH system) and enzymatic method (xanthine oxidase system). DPPH radical scavenging effect of 7, 8, 4'-trihydroxyisoflavone was similar with vitamin C in a dose-dependent manner. In xanthine oxidase (XO) system 7, 8, 4'-trihydroxyisoflavone showed superoxide radical inhibition activity of 50 % at a concentration of $6.6{\pm}0.4\;{\mu}M$. Also, the compound significantly suppressed cellular MMP-1 formation. These results suggest that 7, 8, 4'-trihydroxyisoflavone could be developed as a potential preventive or therapeutic agent against skin aging.

Water Soluble Cyclosporine Monomethoxy Poly(ethyleneglycol) Conjugates as Potential Prod rugs

  • Cho, Hoon;Chung, Yong-Seog
    • Archives of Pharmacal Research
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    • v.27 no.6
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    • pp.662-669
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    • 2004
  • The highly water-soluble monomethoxypoly(ethyleneglycol) (mPEG) prod rugs of cyciosporin A(CsA) were synthesized. These prod rugs were prepared by initially preparing intermediate in the form of carbonate at the 3'-positions of CsA with chloromethyl chloroformate, in the pres-ence of a base to provide a 3'-carbonated CsA intermediate. Reaction of the CsA intermediate with mPEG derivative in the presence of a base provides the desired water-soluble prod rugs. As a model, we chose molecular weight 5 kDa mPEG in the reaction with CsA to give water soluble prodrugs. To prove that the prod rug is decomposed in the body to produce CsA, the enzymatic hydrolysis test was conducted using human liver homogenate at $37^{\circ}C$. The prodrug was decomposed in human liver homogenate to produce the active material, CsA, and the hydrolysis half-life ($t_{1/2}$) of the prodrug, KI-306 was 2.2 minutes at $37^{\circ}C$. However, a demon-stration of non-enzymatic conversion in pH 7.4 phosphate buffer was provided by the fact that the half-life ($t_{1/2}$) is 21 hours at 37$^{\circ}C$. The hydrolysis test in rat whole blood was also conducted. The hydrolysis was seen with half-life ($t_{1/2}$) of about 9.9, 65.0, 14.2, 3.4, 2.1 9.5, and 1.6 minutes for KI-306, 309, 312, 313, 315, 316, and 317, respectively. This is the ideal for CsA prodrug. The pharmacokinetic study of the prodrug, KI-306, in comparison to the commer-cial product (Sandimmune Neoral Solution) was also carried out after single oral dose. Each rat received 7 mg/kg of CsA equivalent dose. Especially, the prodrug KI-306 exhibits higher AUC and $C_{max}$ than the conventional Neoral. The AUC and $C_{max}$ were increased nearly 1.5 fold. The kinetic value was also seen with $T_{max}$ of about 1.43 and 2.44 hours for KI-306 and Neoral, respectively.

Antioxidant Activities and Whitening Effect from Lindera obtusiloba BL. Extract (생강나무 추출물의 항산화 활성과 미백효과)

  • Bang, Chae-Young;Won, Eun-Kyung;Park, Kuen-Woo;Lee, Gwang-Won;Choung, Se-Young
    • YAKHAK HOEJI
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    • v.52 no.5
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    • pp.355-360
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    • 2008
  • In this study we investigated antioxidant activity of against several free radicals and skin whitening effect of 70% ethanol extract (leaf extracts and branch/stem mixed) of Lindera obtusiloba BL. Antioxidant activity was assessed by DPPH, superoxide radical and hydroxyl radical assays. The Lindera obtusiloba BL. extract had antioxidant activity dose dependently with an ${IC}_{50}$ value of 243.14 and 181.10 ${\mu}g$/ml for DPPH, 165.77 and >1500 ${\mu}g$/ml for non-enzymatic system of superoxide radical assay, 35.47 and >100 ${\mu}g$/ml for enzymatic system of superoxide radical assay, 1.21 mg/ml for hydroxyl radical assay. In addition we tested tyrosinase inhibition activity and melanin contents on B16 melanoma F10. B16 melanoma cell was treated by such sample as 1, 5, 10 and 50 ${\mu}g$/ml for 72 hr and tyrosinase inhibition was tested. Melanogenesis was inhibited to 22% at the dose of 50 ${\mu}g$/ml and tyrosinase was inhibited to 45.2% at the same dose. In conclusion Lindera obtusiloba BL had potent antioxidant activity and inhibitory activity of tyrosinase and melanin formation. It could be developed as the health functional food and functional cosmetic resources.

Anti-arthritic activity of D-carvone against complete Freund's adjuvant-induced arthritis in rats through modulation of inflammatory cytokines

  • Chen, Guifang;Song, Yuxiu;Ma, Fang;Ma, Yuxia
    • The Korean Journal of Physiology and Pharmacology
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    • v.24 no.6
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    • pp.453-462
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    • 2020
  • Chronic joint pain due to loss of cartilage function, degradation of subchondral bone, and related conditions are common plights of an arthritis patient. Antioxidant compounds could solve the problems in arthritic condition. The objective of this study was to evaluate the anti-arthritic activity of D-carvone against complete Freund's adjuvant (CFA)-induced arthritis in rats. D-carvone was orally administered for 25 days at the doses of 30 and 60 mg/kg against CFA-induced arthritic rats. Changes in body weight, paw swelling, organ index, hematological parameters, oxidative stress markers, inflammatory cytokines, and histopathology were recorded. Oral treatment of D-carvone significantly improved the body weight, reduced the paw swelling, edema formation, and organ index in arthritic rats. The levels of white blood cells were reduced, red blood cells and hemoglobin levels were improved in D-carvone treated arthritic rats. Lipid peroxidation levels were lowered whereas enzymatic and non-enzymatic antioxidants were significantly elevated by D-carvone administration against arthritic rats. D-carvone significantly modulated inflammatory cytokine levels and improved the ankle joint pathology against CFA-induced arthritic inflammation. In conclusion, D-carvone proved significant anti-arthritic activity against CFA-induced arthritis in rats.

Effect of Microalgal Extracts of Tetraselmis suecica against UVB-Induced Photoaging in Human Skin Fibroblasts

  • Jo, Wol Soon;Yang, Kwang Mo;Park, Hee Sung;Kim, Gi Yong;Nam, Byung Hyouk;Jeong, Min Ho;Choi, Yoo Jin
    • Toxicological Research
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    • v.28 no.4
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    • pp.241-248
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    • 2012
  • Exposure of cells to ultraviolet B (UVB) radiation can induce production of free radicals and reactive oxygen species (ROS), which damage cellular components. In addition, these agents can stimulate the expression of matrix metalloproteinase (MMP) and decrease collagen synthesis in human skin cells. In this study, we examined the anti-photoaging effects of extracts of Tetraselmis suecica (W-TS). W-TS showed the strongest scavenging activity against 2,2-difenyl-1-picrylhydrazyl (DPPH) and peroxyl radicals, followed by superoxide anions from the xanthine/xanthine oxidase system. We observed that the levels of both intracellular ROS and lipid peroxidation significantly increased in UVB-irradiated human skin fibroblast cells. Furthermore, the activities of enzymatic antioxidants (e.g., superoxide dismutase) and the levels of non-enzymatic antioxidants (e.g., glutathione) significantly decreased in cells. However, W-TS pretreatment, at the maximum tested concentration, significantly decreased intracellular ROS and malondialdehyde (MDA) levels, and increased superoxide dismutase and glutathione levels in the cells. At this same concentration, W-TS did not show cytotoxicity. Type 1 procollagen and MMP-1 released were quantified using RT-PCR techniques. The results showed that W-TS protected type 1 procollagen against UVB-induced depletion in fibroblast cells in a dose-dependent manner via inhibition of UVB-induced MMP-1. Taken together, the results of the study suggest that W-TS effectively inhibits UVB-induced photoaging in skin fibroblasts by its strong anti-oxidant ability.

The Principle and Trends of CRISPR/Cas Diagnosis (CRISPR/Cas 진단의 원리와 현황)

  • Park, Jeewoong;Kang, Bong Keun;Shin, Hwa Hui;Shin, Jun Geun
    • Journal of Biomedical Engineering Research
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    • v.42 no.3
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    • pp.125-142
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    • 2021
  • The POCT (point-of-care test) sensing that has been a fast-developing field is expected to be a next generation technology in health care. The POCT sensors for the detection of proteins, small molecules and especially nucleic acids have lately attracted considerable attention. According to the World Health Organization (WHO), the POCT methods are required to follow the ASSURED guidelines (Affordable, Sensitive, Specific, User- friendly, Robust and rapid, Equipment-free, Deliverable to all people who need the test). Recently, several CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) based diagnostic techniques using the sensitive gene recognition function of CRISPR have been reported. CRISPR/Cas (Cas, CRISPR associated protein) systems based detection technology is the most innovative gene analysis technology that is following the ASSURED guidelines. It is being re-emerged as a powerful diagnostic tool that can detect nucleic acids due to its characteristics that enable rapid, sensitive and specific analyses of nucleic acid. The first CRISPR-based diagnosis begins with the discovery of the additional function of Cas13a. The enzymatic cleavage occurs when the conjugate of Cas protein and CRISPR RNA (crRNA) detect a specific complementary sequence of the target sequence. Enzymatic cleavage occurs on not only the target sequence, but also all surrounding non-target single-stranded RNAs. This discovery was immediately utilized as a biosensor, and numerous sensor studies using CRISPR have been reported since then. In this review, the concept of CRISPR, the characteristics of the Cas protein required for CRISPR diagnosis, the current research trends of CRISPR diagnostic technology, and some aspects to be improved in the future are covered.

Improvement of protein extraction efficiency from defatted sesame meal with thermal and enzymatic treatments (열 처리와 효소 처리에 의한 탈지 참깨박 단백질의 추출율 향상)

  • In, Man-Jin
    • Journal of Applied Biological Chemistry
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    • v.63 no.4
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    • pp.291-295
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    • 2020
  • In order to increase the utilization of defatted sesame meal (DSM), a by-product of sesame oil production, the conditions of extraction of insoluble proteins from DSM by enzyme treatment were investigated. As a result of comparing the treatment results of proteolytic enzymes Alcalase, Flavorzyme, Neutrase, and Protamex with control, Protamex was effective in increasing the total solid and protein content. At the reaction conditions of Protamex (50 ℃, pH 6.0), the dosage of enzymes was appropriate for 1% of DSM and 3 h of enzyme reaction time. To improve the efficiency of enzymatic treatment, the protein content extracted increased as the heat treatment temperature increased, and slightly increased above 110 ℃. As a result of investigating the effect of the combination treatment of cell lytic enzyme (Tunicase) and protease (Protamex) on protein solubilization, it was most effective to treat the cell lytic enzyme after processing the protease. After heat treatment (110 ℃, 10 min), sequential treatment of Protamex and Tunicase increased the protein content by about 3.5 times (9.85→35.58 mg/mL) of the non-heated control and 2.2 times (15.83→35.58 mg/mL) of the heat treated control.

Effects of Preheated Additives on the Fermentation Quality of Napiergrass Silage

  • Yunus, M.;Ohba, N.;Tobisa, M.;Shimojo, M.;Masuda, Y.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.11
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    • pp.1564-1567
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    • 2001
  • Hydrolysis of plant protein to non-protein nitrogen (N) or ammonia can reduce quality of silage crops. Heating or non-enzymatic browning is a treatment to inhibit this hydrolysis. This experiment was conducted to examine the effects of pre-heated soybean meal and molasses on the fermentation quality of napiergrass silage. The initial growth of napiergrass was harvested at 85 days of age and immediately chopped into about 1 cm length. About 700 g of the grass was ensiled into a laboratory silo (1.0 liter polyethylene container) and incubated for 30 days at room temperature ($28^{\circ}C$). No additives (control), molasses, soybean meal and molasses + soybean meal treatments were prepared. All additives were non-heated or heated in an oven at $150^{\circ}C$ for 30 minutes before ensiling. Molasses was added at 3% on the fresh weight basis and soybean meal was added at 0.5% N, respectively. After opening the silo, pH, total nitrogen (TN), volatile basic nitrogen (VBN), lactic acid (LA), acetic acid (AA), butyric acid (BA) and dry matter (DM) contents were determined. The data were analyzed statistically by analysis of variance. Compared with control, molasses addition significantly decreased pH value, VBN/TN, AA and BA and increased LA production. Soybean meal addition significantly increased TN and VBN/TN of silage. Both molasses and soybean meal addition significantly reduced pH value, AA, and BA and increased DM and LA contents of silage. The heating of additives was only effective to reduce VBN/TN production compared with non-heated additives in soybean meal and soybean meal with molasses addition.

Antioxidant Constituents from the Stem of Sorghum bicolor

  • Kwon, Yong-Soo;Kim, Chang-Min
    • Archives of Pharmacal Research
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    • v.26 no.7
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    • pp.535-539
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    • 2003
  • The EtOAc soluble fraction from the stem of Sorghum bicolor showed a strong free radical scavenging activity. Five major compounds were isolated from this fraction. They were identified by spectral data as methyl ferulate (1), methyl p-hydroxycinnamate (2), p-hydroxybenzaldehyde (3), tricin (4), and quercetin 3,4 -dimethyl ether (5). Among these compounds, 1 exhibited a strong, free radical scavenging activity on 1, 1-diphenyl-2-picrylhydrazyl (DPPH) with an $IC_50$ value of 0.7 $\mu$M. We further studied the effects of these isolated compounds on the lipid peroxidation in rat liver microsomes induced by non-enzymatic method. All five compounds showed anti-lipid peroxidation activity ($IC_50$ values of 0.5, 0.4, 0.3 and 0.3 $\mu$ M, respectively).