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Neuroprotective and Memory Enhancing Effects of Pinelliae rhizoma Extract (반하가 CT105에 의한 신경세포 상해 및 백서의 기억에 미치는 영향)

  • Gang Sang-Yeol;Lee So-Yeon;Yoon Hyeon-Deok;Shin Oh-Chul;Park Chang-Gook;Park Chi-Sang
    • The Journal of Korean Medicine
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    • v.26 no.3 s.63
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    • pp.27-42
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    • 2005
  • Objectives : Alzheimer's disease (AD) is a progressive and fatal neurodegenerative disease characterized by amyloid plaques and neurofibrillary tangles. These plaques are associated with degenerating neuronal processes and consist primarily of fibrillary aggregates of beta-amyloid$ protein, generated from amyloid precursor protein (APP). Another amyloidogenic fragment, the carboxyl terminus (CT) of APP, which is composed of 99-105 amino acid residues containing the complete $A{\beta}$ sequence, also appears to be toxic to neurones. Recent evidence suggest that CT105, carboxy terminal 105 amino acids peptide fragment of APP, may be an important factor causing neurotoxicity in AD. Methods : Although a variety of oriental prescriptions including Pinelliae rhizoma have traditionally been utilized for the treatment of AD, their pharmacological effects and action mechanisms have not yet been fully elucidated. In the present study, we investigated effects of the dichloromethane extract of Pinelliae rhizoma (PINR) on neurotoxicity and the formation of reactive oxygen species (ROS) and nitric oxide (NO) in SK-N-SH cells overexpressed with CT105. In addition, we evaluated its radical scavenging activity and effects on acetylcholinesterase (AChE) activity. Furthermore, effects on cognitive deficits induced by scopolamine treatment in rats were evaluated. Results ; We found in this study that PINR significantly inhibited apoptotic neuronal death induced by CT105 overexpression in SK-N-SH cells. Based on morphological examinations by phase-contrast microscopy, PINR reversed apoptotic changes of CT105-expressed cells. It was also found that PINR significantly promoted neurite outgrowth and inhibited formation of ROS nd NO. PINR was shown to scavenge DPPH radicals and noncompetitively inhibit AChE activity. Furthermore, it reduced scopolamine-induced memory impairment in rata, assessed by passive avoidance test. Conclusions : Taken together, these results demonstrate that PINR exhibits neuroprotective, antioxidant, and memory enhancing effects, and therefore may bs beneficial for the treatment of AD.

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Simultaneous determination of baicalin and glycyrrhizin in Sosiho-tang by HPLC and anti-inflammatory activity (소시호탕에서 baicalin과 glycyrrhizin 동시 분석 및 항염 활성)

  • Kim, Jung-Hoon;Ha, Hye-Kyung;Seo, Chang-Seob;Lee, Jin-Ah;Shin, Hyeun-Kyoo
    • The Korea Journal of Herbology
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    • v.26 no.1
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    • pp.41-46
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    • 2011
  • Objectives : To quantitate the main compounds and investigate the biological activity of Sosiho-tang (Xiao-Chai-Hu-Tang, SST), simultaneous determination of baicalin and glycyrrhizin, and anti-inflammatory activity were estimated. Methods : A quantitative analysis was performed using a high performance liquid chromatography (HPLC). Reference compounds were separated on a reversed-phase column using gradient elution with water and acetonitrile each containing acetic acid at a flow rate of 1 mL/min. And the productions of nitric oxide (NO) and prostaglandin $(PE)E_2$ were examined by lipopolysaccharide (LPS)-treated RAW 264.7 cells in the presence of the SST. The anti-inflammatory activity of SST was investigated by carrageenin-induced paw edema in rats. The paw volume was measured at 2 and 4 hr following carrageenin-induced paw edema in rats. Results : The correlation coefficients of the compounds showed good linearity ($r^2$ > 0.9992) over the linear range. The precisions of intra- and inter-day were less than 7.0% of relative standard deviation (RSD) values for baicalin and less than 3.5% of RDS valuse for glycyrrhizin. Recovery rates were within the range of 95.41-101.5%. The contents of baicalin and glycyrrhizin in SST were average 70.52, 6.18 mg/g, respectively. And SST exhibited inhibitory effect on NO production in LPS-treated RAW 264.7 cells but not on $PGE_2$ production. Oral administration of SST (1 g/kg) showed a reduction in carrageenin-induced paw edema on rats. Conclusions : The analytical method was applied successfully to measure the contents of baicalin and glycyrrhizin in SST which exhibited anti-inflammatory activities.

Proteomic analysis of Korean mothers' human milk at different lactation stages; postpartum 1, 3, and 6 weeks (출산 후 경과한 날에 따른 한국인 산모의 모유 단백체 분석)

  • Park, Jong-Moon;lee, Hookeun;Song, Seunghyun;Hahn, Won-Ho;Kim, Mijeong;Lee, Joohyun;Kang, Nam Mi
    • Analytical Science and Technology
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    • v.30 no.6
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    • pp.348-354
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    • 2017
  • In this study, patterns of proteome expression were monitored and specifically expressed proteins in human milk were detected in collected human milk after 1 week, 3 weeks, and 6 weeks from delivery. A quantitative shotgun proteomic approach was used to identify human milk proteins and reveal their relative expression amounts. For each sample, two independent human milk samples from two mothers were pooled, and then three replicated shotgun proteomic analyses were carried out. Casein, which is a highly abundant protein in human milk, was removed, and then trypsin was treated to produce a digested peptide mixture. The peptides were loaded in the home-made reversed-phase C18 fused-silica capillary column, and then the eluted peptides were analyzed by using a linear ion-trap mass spectrometer. The relative quantitation of proteins was performed by the normalized spectral count method. For each sample, 81-109 non-redundant proteins were identified. The identified proteins consisted of glycoproteins, metabolic enzyme, and chaperon enzymes such as lactoferrin, carboxylic ester hydrolase, and clusterin. The comparative analysis for the 63 proteins, which were reproducibly identified in all three replications, revealed that 25 proteins were statically significant differentially expressed. Among the differentially expressed proteins, Ig lambda-7 chain C region and tenascin drastically decreased with the delivery time.

Induction of Apoptosis by Citri Pericarpium Methanol Extract through Reactive Oxygen Species Generation in U937 Human Leukemia Cells (진피 메탄올 추출물의 활성산소종 생성을 통한 인체 백혈병 세포의 apoptosis 유발)

  • Kim, Ga Hee;Lee, Moon Hee;Han, Min Ho;Park, Cheol;Hong, Su Hyun;Choi, Yung Hyun
    • Journal of Life Science
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    • v.23 no.8
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    • pp.1057-1063
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    • 2013
  • Citri Pericarpium is one of the most commonly used traditional herbal medicines in Korea, China, and Japan. Its extracts have many properties including the treatment of indigestion and inflammatory respiratory syndromes such as bronchitis and asthma. However, the underlying molecular mechanisms of anti-cancer activity and molecular targets are not fully understood. In this work, we investigated the anti-proliferative activity of Citri Pericapium (EMCP) methanol extract on reactive oxygen species (ROS) production and the association of these effects with apoptotic cell death using U937 human leukemia cells in vitro. EMCP treatment decreased cell proliferation in a dose-dependent manner following an increase of the sub-G1 phase, the down-regulation of Bax proteins, the activation of caspases, the degradation of poly (ADP-ribose) polymerase proteins (PARP), and the induction of ROS generation. However, the quenching of ROS generation by N-acetyl-L-cysteine administration, a scavenger of ROS, reversed the EMCP-induced apoptosis effects. In addition, heme oxygenase-1 expression also recovered by inhibiting the nuclear translocation of phosphorylated NF-E2-related factor 2. Taken together, our data indicate that ROS are involved as key mediators in the early molecular events in the EMCP-induced apoptotic pathway.

Development of Vitamin D Determination in Infant Formula by Column-Switching HPLC with UV Detector

  • Ko, Jin-Hyouk;Kwak, Byung-Man;Ahn, Jang-Hyuk;Shim, Sung-Lye;Kim, Kyong-Su;Yoon, Tae-Hyung;Leem, Dong-Gil;Jeong, Ja-Young
    • Food Science of Animal Resources
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    • v.32 no.5
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    • pp.571-577
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    • 2012
  • This study was carried out to develop an analytical method for the determination of vitamin D in infant formula. Vitamin D was determined by column-switching high-performance liquid chromatography (HPLC) equipped with a reversed phase column and UV detector after saponification and extraction of the formula with an organic solvent. A preseparation column ($C_8$), focusing column ($C_{18}$), analytical column ($C_{18}$) and UV-Vis detector (254 nm) were used. The limits of detection (LOD) and the limits of quantification (LOQ) for vitamin D were estimated to be $1.51{\mu}g/kg$ and $4.95{\mu}g/kg$, respectively. The linearity, recovery, precision and accuracy of the analytical method for vitamin D were evaluated through the application of a SRM (Standard Reference Material) 1846 (National Institute of Standard & Technology, USA). The linearity of this method was calculated with a value of the coefficient of determination ($r^2$) ${\geq}0.9999$. The recovery of vitamin D was $85.20{\pm}3.00%$. The intra-assay precision for vitamin D was between $1.68{\pm}0.03%$ and $5.75{\pm}0.33%$, and the inter-assay precision for vitamin D ranged from $1.73{\pm}0.03%$ to $2.96{\pm}0.09%$. The intra-assay accuracy for vitamin D was between $100.03{\pm}2.77%$ and $102.01{\pm}0.59%$, and the inter-assay accuracy for vitamin D ranged from $99.00{\pm}1.53%$ to $102.01{\pm}3.04%$. The proposed method is optimal for the separation and quantification of vitamin D from infant formula.

Chemical Structures and Physiological Activities of Plant Growth Substance, Malformin A's (식물생장조절물질 말포민 A동족체의 화학구조 및 생리활성)

  • Kim, K.W.
    • Korean Journal of Weed Science
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    • v.15 no.1
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    • pp.73-84
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    • 1995
  • Four malformin A's produced by Aspergillus niger van Tiegh. were separated by HPLC equipped with $C_{18}$ reversed-phase column and subjected to structural determination. Amino acid analyses and mass spectra data of the compounds indicate that they structurally resemble the cyclic pentapeptide malformin $A_1$. Their structures were deduced by two dimensional NMR and MS/MS experiments as cyclo-D-Cys-D-Cys-L-Val-D-Leu-L-Ile for $A_1$, cyclo-D-Cys-D-Cys-L-Val-D-Leu-L-Val for $A_2$, cyclo-D-Cys-D-Cys-L-Val-D-Leu-L-Leu for $A_3$, and cyclo-D-Cys-D-Cys-L-Val-D-Ile-L-Val for $A_4$. Among the mal-formin A's, the structure of $A_3$ was identical to that of malformin C, which was produced by A. niger strain AN-1. All the malformin A's caused severe curvatures of corn(Zea mays L.) roots and the activities of the malformin A's with molecular weight 529 were greater than those with molecular weight 515. Malformin $A_1$ caused the corn root curvature by 83% at a concentration of $0.25{\mu}M$. In the mung bean(Phaseolus aureus Roxb.) hypercotyl segment test, however, the molecular weight of malformin A's was not a factor influencing the physiological activities. Malformin $A_1$ stimulated the growth of mung bean hypercotyles by 165% at a $0.1{\mu}M$ concentration.

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Determination of alkylphenol ethoxylate in water by high performance liquid chromatography/electrospray ionization/mass spectrometry (HPLC/ESI/MS를 이용한 물 중의 알킬페놀에톡실레이트 분석)

  • Lee, Jeongae;Park, Song-Ja;Chung, Bong Chul
    • Analytical Science and Technology
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    • v.17 no.3
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    • pp.263-270
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    • 2004
  • A method is described for the analysis of short-chain alkylphenol ethoxylates (APEOs), 4-octylphenol-di-ethoxylate (OP2EO) and 4-nonylphenol-di-ethoxylate (NP2EO), in drinking water or wastewater using reversed phase high-performance liquid chromatography with electrospray ionization mass spectrometry. The solvent system was water and methanol containing $10{\mu}M$ trifluoroacetic acid as an ionization solvent. We acidified 1 L of water samples to less than pH 2 with concentrated $H_2SO_4$ and loaded onto Sep-Pak $C_{18}$, and eluted with acetone. The calibration of OP2EO and NP2EO was performed for the concentration range from 20 to 500 ng/L and the correlation coefficients were 0.999 and 0.990, respectively. The limits of detection were 20 ng/L (OP2EO) and 50 ng/L (NP2EO) at a signal-to-noise ratio of 3. Accuracy and precision of this analytical method were 85.8 ~ 122.1% and 8.2 ~ 18.8%, respectively. The proposed method allowed a sensitive and rapid detection of OP2EO and NP2EO and it could be applied for monitoring of APEOs from environmental samples.

A COMPARISON OF RELEASE RATE AND CUMULATIVE RELEASE OF TEGDMA WITH OR WITHOUT THE APPLICATION OF BONDING RESIN (접착레진 적용 유무에 따른 TEGDMA의 방출속도 및 방출량 비교)

  • Shin, Hee-Jung;Jeon, Seong-Min
    • Restorative Dentistry and Endodontics
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    • v.23 no.2
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    • pp.701-709
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    • 1998
  • Many dental composites are Bis-GMA based resin which diluted with the more fluid monomer triethylenglycol dimethacrylate(TEGDMA). TEGDMA is often present in exess so that some quantity remains unreacted following photo-initiated polymerization. TEGDMA is a component of some resin composites which contributes to their cytotoxicity. The presence of dentin between resin composite and pulp space reduce the cytotoxicity in vitro. The root system from extrcted human third molar was removed and then a circular occlusal cavity 4mm in diameter was prepared, leaving a remaining dentinal thickness to the roof of the pulpal chamber within the range 1.0-1.5mm. Dentine was treated with 37% phosphoric acid prior to Z 100 placement without using bonding resin(group 1). In group 2, SMP(Scotchbond Multi Purpose) primer, bonding resin prior to Z 100 placement were applied sequently. In group 3, moulds with internal dimensions 4mm diameter by 2mm depth were used to contain the composite alone with an equvalent mass on tooth model, and then they were immersed directly into water. The purpose of this study is to evaluate the release rate and quantity of TEGDMA with or without the application of bonding resin. Both release rate and total cumulative amount of TEGDMA for the three groups were determined using reversed-phase HPLC at times up to 10 days. The results were as follows: 1. All experimental groups showed the highest rate of release was in the first sample period(0-4.32 min) and the rate of release declined exponentially thereafter. 2. The maximum release rate and total cumulative account of TEGDMA in the tooth model of group 1 and group 2 with the use of SMP bonding resin were reduced however ther were no significant differences between these groups(P>0.05). 3. In the first sample period(0-4.32 min), the rate of release of TEGDMA from composite resin in group 3 immersed directly into water was significantly higher than that in group 1 and group 2 of tooth model(P<0.05). Conclusively, TEGDMA diffusion from Z 100 resin was not effectively prevented by the presence of dentin in spite of using the SMP bonding resin.

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Purification and Characterization of a Novel Neuvopeptide with a Smooth Muscle-Relaxing Activity from the Starfish, Asterina pectinifera (별불가사리(Asterina pectinifera)로부터 새로운 평활근 이완활성 펩타이드의 정제)

  • KIM Eun Jung;KIM Chan-Hee;GO Hye-Jin;KIM In Hae;AN Sang Hyun;SOHN Hee-Young;PARK Hee Yeon;YOON Ho Dong;CHANG Young-Chae;HONG Yong-Ki;PARK Nam Gyu
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.38 no.3
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    • pp.148-152
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    • 2005
  • A novel neuropeptide with a relaxing activity on the dorsal retractor muscle (DRM) was isolated from the whole body extract of the starfish, Asterina pectinifera. The peptide was purified by gel-filtration ion-exchange and $C_{18}$ reversed-phase HPLC. The complete amino acid sequence of this peptide, which was determined by automated Edman degradation and MALDI-TOF mass, was Phe-Gly-Lys-Gly-Gly-Ala-Tyr-Asp-Pro-Leu-Ser-Ala-Gly-Phe-Thr-Asp. A comparison of the amino acid sequence with those of other known neuropeptides revealed that the asteripectin was a novel neuropeptide with smooth muscle-relaxing activity on the starfish DRM. This peptide showed threshold response to relaxing activity on the DRM at $10^{-10}M$ and the maximal relaxing effect was $120{\pn}7.0\%$ at $10^{-5}M$. The relaxing activity of this peptide on the starfish DRM increased in a dose-dependent manner.

Enzymatic Synthesis of Low-trans Fats Containing Conjugated Linoleic Acids and Their Physicochemical Characteristics (Conjugated Linoleic Acid(CLA)를 함유한 기능성 저트랜스 유지의 효소적 합성 및 이화학적 특성 연구)

  • Nam, Ha-Young;Lee, Ki-Teak
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.6
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    • pp.752-760
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    • 2008
  • Scale-up production of low-trans fat containing conjugated linoleic acid (CLA-TFO) was performed through lipase-catalyzed synthesis. Blend of fully hydrogenated soybean oil, olive oil containing conjugated linoleic acid and palm oil with 1:2:7 ratio was interesterified through Lipozyme RM IM in the 1 L-batch type reactor at $65^{\circ}C$ for 12 hrs, and the physicochemical and melting properties of CLA-TFO were compared with conventional (high trans fat) or commercial low-trans fat shortening. The trans fatty acids content in the conventional shortening (48.8 area%) was much higher than that of low-trans shortening (0.4 area%) and CLA-TFO (0.3 area%+CLA; 7.6 area%). Acid, saponification and iodine values of CLA-TFO were 0.4, 173.9 and 59.0, respectively. Their ${\alpha}$-, ${\gamma}$-tocopherol contents showed 4.7, 1.0 mg/100 g. Differences were observed in the solid fat contents (SFC), melting point of the conventional or low-trans fat and CLA-TFO. Each SFC of conventional, low-trans fat and CLA-TFO was 32.0, 29.3 and 30.4% with melting point of 38.5, 43.0 and $39.5^{\circ}C$ at $35^{\circ}C$, respectively. In texture profile analysis, hardness of conventional, low-trans fat and CLA-TFO was 111.7, 75.2 and 63.8 g.