• Title/Summary/Keyword: AP-l

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Expression and Purification of an ACE-Inhibitory Peptide Multimer from Synthetic DNA in Escherichia coli

  • OH, KWANG-SEOK;YONG-SUNG PARK;HA-CHIN SUNG
    • Journal of Microbiology and Biotechnology
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    • v.12 no.1
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    • pp.59-64
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    • 2002
  • An angiotensin I-converting enzyme (EC 3.4.15.1) (ACE), which can convert inactive angiotensin I into angiotensin II, a vasoconstrictor, is one of the key enzymes in controlling hypertension. It is suggested that the inhibition of ACE prevents hypertension, and many inhibitory peptides have already been reported. In the current study, oligonucleotides encoding ACE inhibitory peptides (IY, VKY) were chemically synthesized and designed to be multimerised due to isoschizomer sites (BamHI, BglII). The cloned gene named AP3 was multimerised up to 6 times in pBluescript and expressed in BL2l containing pGEX-KG. The fusion protein (GST-AP3) was easily purified with a high recovery by an affinity resin, yielding 38 mg of synthetic AP3 from a 1-1 culture. The digestion of AP3 by chymotrypsin exhibited an $IC_50$ value of $18.53{\mu}M$. In conclusion, the present experiment indicated that AP3 could be used as a dietary antihypertensive drug, since the potent ACE inhibitory activity of AP3 could be activated by chymotrypsin in human intestine.

Ethanol-induced Activiationof Transcription Factor NF-$\kappa$B and AP-1 in C6 Glial Cells

  • Park, Jae -Won;Shim, Young-Sup
    • Preventive Nutrition and Food Science
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    • v.4 no.3
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    • pp.209-214
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    • 1999
  • In this study, the effectof ethanol and acetaldehyde on DNA binding activities of NF-$textsc{k}$B and AP-1 were evaluated in C6 rat glial cells. Both NF-$textsc{k}$B and AP-1 are important transcription factors for the expression of various cytokines in glial cells. Our data showed that neither ethanol nor acetaldehyde induced conspicuous cell death of C6 cells at clinically realistic concentrations. When the DNA binding activities of nuclear NF-$textsc{k}$B and AP-1 were estimated using electrophoretic mobility shift assay (EMSA), ethanol(0.3%) or acetaldehyde(1mM) induced transient activation of these transcription factors, which attained peak levels at 4~8 hours and declined to basal levels at 12 hours after treatement . The supershift analysis showed that the increased activities of NF-$textsc{k}$B in ethanol/acetaldehyde-treated C6 cells were due to the preferential induction of p65/p50 heterodimer complex. The DNA binding activities of these transcriptional factors decreased below basal levels when cells were cultured with either ethanol or acetaldehyde for 24 hours, and showed the inhibitory effect of chronic ehtanol /acetaldehyde treatment on the activities of these transsriptional factors. Our data indicate that either ethanol or acetaldehyde can induce functional changes of glial cells throught bi-directional modulation of NF-$textsc{k}$B and AP-1 DNA binding activities.

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Validation of HPLC Methods for Ascorbic Acid and Its Derivatives in Foods (식품 중 아스코르빈산 유래 산화방지제의 HPLC 분석법 검증 및 개선)

  • Jeong, Min Kyu;Park, Chan Uk;Park, Min Hee;Yeo, JuDong;Park, SeungKwan;Kim, SoHee;Shin, Tae-Sun;Baek, Hyung Hee;Lee, JaeHwan
    • Food Engineering Progress
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    • v.15 no.1
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    • pp.75-79
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    • 2011
  • Analytical methods for food antioxidants including ascorbic acid, erythorbic acid, ascorbyl palmitate (AP), and ascorbyl stearate (AS), were validated using high performance liquid chromatography. Validation parameters such as linearity, limit of detection (LOD), limit of quantification (LOQ), and recovery were tested using lard and cider as food model systems. Linearity of ascorbic acid and erythorbic acid were both higher than ($R^2$> 0.99), LOD of these compounds were 0.46 and 0.48 ${\mu}g/mL$, respectively and LOQ were 1.39 and 1.45 ${\mu}g/mL$, respectively. The recovery rates of these compounds were 86.35-94.78% and 84.76-95.02%, respectively. However, the concentration of AP and AS decreased in methanol stock solution. Four other solvents including ethanol, acetonitrile, mixture of methanol and acetonitrile, and mixture of ethanol and acetonitrile were tested to increase the stability of AP and AS under room temperature and refrigerated temperature. Ethanol provided better stability of AP and AS under both room and refrigerated temperature. This study can help to accurately analyze the content of ascorbic acid and its derivatives in processed foods.

Antioxidative and Antibacterial Activities of Artemisia princeps Pampanini Extracts (사자발쑥 추출물의 항산화 및 항균 활성)

  • Yang, Hyun Gab;Kim, Hye Jin;Kim, Hae Soo;Park, Soo Nam
    • Microbiology and Biotechnology Letters
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    • v.40 no.3
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    • pp.250-260
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    • 2012
  • In the present study, the antioxidative and antibacterial activities of Artemisia princeps Pampanini (A. princeps Pamp.) extract were investigated. The ethyl acetate fraction of A. princeps Pamp. showed the most prominent free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activity ($FSC_{50}=12.27{\mu}g/mL$). Reactive oxygen species (ROS) scavenging activities ($OSC_{50}$) of A. princeps Pamp. extract on $Fe^{3+}-EDTA/H_2O_2$ systems were investigated using a luminol-dependent chemiluminescence assay. The ethyl acetate fraction of the extract ($OSC_{50}=0.33{\mu}g/mL$) had a 5 times greater ROS scavenging activity than L-ascorbic acid ($1.50{\mu}g/mL$), known as a water soluble antioxidant. The cellular protective effects of fractions of A. princeps Pamp. on the rose-bengal sensitized photohemolysis of human erythrocytes were examined. The aglycone fraction of extracts suppressed photohemolysis in a concentration dependent manner. The inhibitory effects of A. princeps Pamp. extract on tyrosinase were investigated to assess their whitening efficiency. The ethyl acetate fraction demonstrated a 7 times higher tyrosinase inhibitory effect ($IC_{50}=29.20{\mu}g/mL$) than albutin, known as a whitening agent. The antibacterial activity of ethyl acetate fractions against various normal skin flora were measured. The results showed that the antibacterial activity of the fraction was the highest on Staphylococcus aureus, Bacillus subtilis, and Propionibacterium acnes. Antioxidant substances were isolated and purified from the ethyl acetate fractions. Eupatilin and jaceosidin were identified. These results indicate that the extract/fractions of A. princeps Pamp. can function as antioxidant and/or antibacterial agents for the skin.

Epidemiological Investigation of Methicillin-Resistant Staphylococcus aureus by Arbitrarily Primed PCR

  • Yang Byoung-Seon
    • Biomedical Science Letters
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    • v.10 no.4
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    • pp.473-477
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    • 2004
  • Methicillin-Resistant Staphylococcus aureus (MRSA) strains are resistant to a wide range of antibiotics and are a major cause of nosocomial infections. Accurate and rapid typing of MRSA is needed to implement effective infection control measures. Arbitrarily Primed PCR (AP-PCR) is a very useful method in rapid typing. AP-PCR is not necessary information about target DNA sequence because this is basically DNA amplification and could be useful in epidemiological typing by classified band pattern. In this study, MRSA were isolated and identified from ICU, Neu, IM and Ped environments and investigated molecular typing by AP-PCR. Ped, the MRSA pattern determines the la, IIa type, 1M is Ib type, Neu is IIa type and ICU determines the IIa, lIb types. All MRSA in this study were typeable by AP-PCR, which was easy to perform and reproduce with evidence of MRSA for purposes of nosocomial infection control.

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Synthesis, ESR and Electrochemical Characterization of Dioxygen Binding to Dirhodium Complexes with 2-anilinopyridinato Bridging Ligand (2-아닐리노 피리딘을 배위자로 하는 이핵 로듐착물의 두 산소첨가 생성물에 대한 합성 및 전기화학적 성질)

  • Kwang Ha Park;Moo Jin Jun;John. L. Bear
    • Journal of the Korean Chemical Society
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    • v.33 no.6
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    • pp.633-643
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    • 1989
  • The R$Rh_2(ap)_4$(2,2-trans) isomer (ap = 2-anilinopyridinate), which has two anilino nitrogens and two pyridyl nitrogens bound to each rhodium ion trans to their own kind, shows activation towards the one electron reduction of dioxygen at -0.40 V vs SCE. The ESR spectrum taken at 123 K proves the formation of a $[Rh_2(ap)_4(O_2)]$ ion with oxygen axially bound to one rhodium ion and the complex is at a RhⅡ2 oxidation state. The complex will form [$Rh_2(ap)_4(O_2)(CH_3CN)]^-$ in presence of $CH_3CN/CH_2Cl_2$ mixture without breaking the Rh-$O_2^-$ bond. When oxidized at -0.25 and 0.55 V, $[Rh_2(ap)_4(O_2)]$ will undergo two one electron oxidations to form $Rh_2(ap)_4(O_2)[Rh_2(ap)_4(O_2)]^+$. Both species have an axially bound superoxide ion but the former is at $Rh^{II}Rh^{III }$and the later at $Rh^{III}_2$ oxidation states. The ESR spetra and $CH_3CN$ addition study, on the other hand, show that the later complex is better described as $[Rh_{II}Rh^{III}(ap)_4(O_2)]^+$ with the odd electron localized on rhodium ion and the complex has an axially coordinated molecular oxygen. The electrochemical and ESR studies also show that the degree of dioxygen activation is a function of electrochemical redox potential.

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The Effects of Robo-horseback Riding with Changes of Pelvic Tilting and Speeds on Muscle Activities of Trunk and Lower Limb (승마기구에서 골반자세와 속도가 몸통 및 다리 근 활성에 미치는 영향)

  • Kim, Jung-Hyun;Chang, Soo-Kyung
    • The Journal of Korean Physical Therapy
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    • v.26 no.5
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    • pp.290-295
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    • 2014
  • Purpose: The purpose of this study was to assess the effects of robo-horseback riding with changes of pelvic tilting and speeds on muscle activities of trunk and lower limb in healthy adults. Methods: Thirty healthy adults were recruited for this study. Subjects performed robo-horseback riding at three pelvic postures (anterior tilting pelvic posture: AP, neutral pelvic posture: NP, posterior pelvic posture: PP) and different speeds (moderate and fast speed). Muscle activities were recorded from the trunk muscles (rectus abdominis [RA], T2 paraspinalis [T2 PS], L2 paraspinalis [L2 PS]) and lower limb muscles (rectus femoris [RF], vastus medialis [VM], vastus lateralis [VL], biceps femoris [BF], gastrucnemius [GCM]). Results: Significantly higher RA activities were observed under the PP condition than under the AP and NP conditions at both speeds. Significantly higher L2 PS activities were observed under the AP condition than under the PP and NP conditions at both speeds. Significantly higher T2 PS activities were observed under the AP condition than under the PP and NP conditions at fast speed. However, the muscle activities of the VM and GCM did not show significant changes with changes in pelvic posture at both speeds. All muscle activities were significantly higher under the fast speed than under the moderate speed in three pelvic postures. Conclusion: There were significant differences in the changes of pelvic tilting on muscle activities of trunk and lower limb were observed in healthy adults. NP condition can be introduced the most efficienct posture for muscle activities of trunk and lower limb in robo-horseback riding.

Alkaline Phosphatase Activity in Two Geologically Different Streams in Alabama, U.S.A. (미국 알라바마에서 지질학적으로 다른 두 하천의 Alkaline Phosphatase 활성도)

  • Joo, Gea-Jae;Ward, Amelia K.
    • The Korean Journal of Ecology
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    • v.18 no.1
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    • pp.1-15
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    • 1995
  • Alakline phosphatase activity (AP A) as a phosphorus deficiency measurement in flowing waters and of microhabitats (rocks, wood, leaves, and sediments) was measured and its relationship to flux of nutrients and response to rainfall events were determined for two geologically different streams in west Alabama from August to November. Results indicated water column AP A in both streams had a low correlation with levels of orthophosphate, total organic phosphorus, nitrate, ammonia, dissolved organic carbon, and discharge (r=0.075-0.583; n=g-IU. Communities on rock surfaces showed a higher AP A level than those on wood and leaves. Sediment passed through a $106{\mu}m$ sieve showed 2-9 times higher AP A level than material passed through $425{\mu}m$ sieve. The first storm after drought at Yellow Creek introduced substantial quantities of DOC (2.5 times baseflow concentrations) and $N0_3-N$ (5.8 times baseflow concentrations) which did not affect AP A significantly. The second storm at Little Schultz Creek caused minor changes in nutrient cocentrations; however $N0_3-N$ levels and AP A were drastically lower due to the dilution effect. Retention of stream water AP A at Yellow Creek and Little Schultz Creek on $0.45{\mu}m$ filter (54 and 43%, respectively) and $0.22{\mu}m$ (83 and 77% of total APA. respectively) indicated more free dissolved portion of the enzyme was present at Little Schultz Creek. Little Schultz Creek (with carbonate and with a higher productivity and biomass) showed a consistantly greater AP A activity $(132{\pm}54\;{\mu}M{\cdot}1^{-1}{\cdot}min^{-I};\;n=g)$ than Yellow Creek $(41{\pm}23\;{\mu}M{\cdot}1^{-I}{\cdot}min^{-I}$, with a sandstone substrate; n=l1, $p{\leq}O.OO1)$. Overall, a greater APA on all microhabitats and the presence of more dissolved enzyme in Little Schultz Creek during the study period may indicates it is more P deficient than Yellow Creek.

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Calcium release and physical properties of modified carbonate apatite cement as pulp capping agent in dental application

  • Zakaria, Myrna Nurlatifah;Cahyanto, Arief;El-Ghannam, Ahmed
    • Biomaterials Research
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    • v.22 no.4
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    • pp.346-351
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    • 2018
  • Background: Carbonate apatite ($CO_3Ap$) and silica-calcium phosphate composite (SCPC) are bone substitutes with good prospect for dental application. SCPC creates a hydroxyapatite surface layer and stimulate bone cell function while, $CO_3Ap$ induce apatite crystal formation with good adaptation providing good seal between cement and the bone. Together, these materials will add favorable properties as a pulp capping material to stimulate mineral barrier and maintain pulp vitality. The aim of this study is to investigate modification of $CO_3Ap$ cement combined with SCPC, later term as $CO_3Ap-SCPC$ cement (CAS) in means of its chemical (Calcium release) and physical properties (setting time, DTS and pH value). Methods: The study consist of three groups; group 1 (100% calcium hydroxide, group 2 $CO_3Ap$ (60% DCPA: 40% vaterite, and group 3 CAS (60% DCPA: 20% vaterite: 20% SCPC. Distilled water was employed as a solution for group 1, and $0.2mol/L\;Na_3PO_4$ used for group 2 and group 3. Samples were evaluated with respect to important properties for pulp capping application such as pH, setting time, mechanical strength and calcium release evaluation. Results: The fastest setting time was in $CO_3Ap$ cement group without SCPC, while the addition of 20% SCPC slightly increase the pH value but did not improved the cement mechanical strength, however, the mechanical strength of both $CO_3Ap$ groups were significantly higher than calcium hydroxide. All three groups released calcium ions and had alkaline pH. Highest pH level, as well as calcium released level, was in the control group. Conclusion: The CAS cement had good mechanical and acceptable chemical properties for pulp capping application compared to calcium hydroxide as a gold standard. However, improvements and in vivo studies are to be carried out with the further development of this material.

Quality Changes of Commercial Kimchi Products by Different Packaging Methods (소포장 김치의 포장방법별 품질특성 변화)

  • Hong, Seok-In;Park, Jin-Sook;Park, Noh-Hyun
    • Korean Journal of Food Science and Technology
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    • v.27 no.1
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    • pp.112-118
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    • 1995
  • The effect of various packaging methods on kimchi quality was investigated in order to develop the packaging techniques for preventing commercial kimchi products from inflation and explosion, due to fermentative gas evolved during storage and distribution. Kimchi was packaged in different methods; 1) atmospheric packaging(AP), 2) check valve packaging(CV), 3) double packaging(DP), and 4) vacuum packaging(VP). The quality of kimchi during storage at $10^{\circ}C$ was evaluated in terms of gas composition, free volume, pH, titratable acidity color index and sensory properties. The gas composition inside packages showed different curves according to the packaging methods. Due to fermentative gas accumulation in both AP and CV, $CO_2$ concentration increased by 2 stepwise pattern, while $O_2$, concentration decreased exponentially. In DP, $O_2$ concentration remained constant, but $CO_2$ concentration increased by 2 stepwise pattern and then decreased. In contrast, VP produced low $O_2$ and high $CO_2$ concentrations only at the end of storage. The free volume in both AP and CV showed typical sigmoidal curves similar to $CO_2$ concentration changes. It remained constant in DP, but started to increase at the late stage of storage in VP. There was no significant effect of packaging methods on pH changes of kimchi. In titratable acidity, DP maintained relatively higher than others. Regarding to the color change of crushed kimchi juice in all packages, L and b values decreased exponentially but a value remained constant during storage. Color index(L b/a) of crushed kimchi juice decreased exponentially and remained constant at the end of storage. The growth of lactic acid bacteria was VP, CV, AP, DP in increasing order. In sensory test, the sourness scores of DP were fairly higher than those of others, but the texture was not significantly affected by the packaging methods. The preference for kimchi showed VP>AP, CV>DP in order of score. In this study, it could be proposed to employ DP and VP method as the effective packaging techniques for preventing commercial kimchi products from inflation.

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