• Title/Summary/Keyword: Chemical assay

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Phenolic compounds from the flowers of Coreopsis lanceolata (큰금계국(Coreopsis lanceolata) 꽃으로부터 phenolic 화합물들의 분리 및 동정)

  • Kim, Hyoung-Geun;Oh, Hyun-Ji;Ko, Jung-Hwan;Jung, Young Sung;Oh, Seon Min;Lee, Yeong-Geun;Kim, Dae-Ok;Lee, Dae Young;Baek, Nam-In
    • Journal of Applied Biological Chemistry
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    • v.62 no.4
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    • pp.323-326
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    • 2019
  • The flowers of Coreopsis lanceolata were extracted with 80% aqueous MeOH and the concentrates were partitioned into EtOAc, n-BuOH, and H2O fractions. The repeated silica gel (SiO2) and octadecyl silica gel column chromatographies for the EtOAc fraction led to isolation of one flavonol and one benzoyl compounds. The chemical structures of the compounds were respectively determined as melanoxetin (1) and protocatechuic acid methyl ester (2) based on spectroscopic analyses including NMR, IR, and MS. These two compounds were isolated for the first time from C. lanceolata flowers in this study. All fractions and the isolated compounds were evaluated for 2,2-diphenyl-1-picrylhydrazyl and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid radical scavenging activities.

Cytotoxic Activity of Bombyx mori and Morus alba Derived Materials against Human Tumor Cell Lines (누에 및 뽕나무 유래 물질의 인간(人間) 암세포주(癌細胞株)에 대한 세포독성(細胞毒性))

  • Park, Il-Kwon;Lee, Jeong-Ock;Lee, Hoi-Seon;Seol, Kwang-Youl;Ahn, Young-Joon
    • Applied Biological Chemistry
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    • v.41 no.2
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    • pp.187-190
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    • 1998
  • The cytotoxic activity of MeOH extracts of the freeze-dried silkworm (Bombyx mori)-derived materials (4th instar larvae, female and mate pupae, virgin female and male adult), dried Beauveria bassiana-infected silkworm larvae, dried feces from the 4th instar larvae B. mori, and dried mulberry (Morus alba)-derived materials (leaves, fruits, root barks) in vitro was evaluated by sulforhodamine B assay, using the five human solid A 549 lung, SK-OV-2 ovarian, SK-MEL-2 melanoma, XF-498 CNS and HCT-15 colon tumor cell lines. The responses varied with both cell line and material used. The 70% hot MeOH extract of B. mori feces (BFH) revealed potent cytotoxic activity against model tumor cell lines whereas moderate activity was observed from the MeOH extract of B. mori feces. M. alba root barks, and M. alba fruits. The other test materials were ineffective. Because of its potent cytotoxic activity, the activity of each solvent fraction from the BFH was determined. Chloroform and ethyl acetate fractions showed the most potent cytotoxic activity. In conclusion, our results may be an indication of at least one of the pharmacological actions of B. mori feces. M. alba root barks, and M. alba fruits.

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Screening and characteristics of anti-complementary polysaccharides from Chinese medicinal herbs (한약재로부터 항보체 활성 다당의 검색 및 특성)

  • Shin, Kwang-Soon;Kwon, Kyung-Sup;Yang, Han-Chul
    • Applied Biological Chemistry
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    • v.35 no.1
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    • pp.42-50
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    • 1992
  • We conducted screening on Chinese medicinal herbs to examine their anti-complementary activity by hemolytic complementary assay $(TCH_{50})$. Among 55 kinds of herbs, several herbs showed relatively potent anti-complementary activity which decreased $TCH_{50}$, more than 70% in comparison with control. Then, hot water extracts of the following herbs, Curcuma aromatica, Areca catechu, Gleditsiae spina, Euonymus alata, Acanthopanax senticous. Lonicera japonica, Aconitum carmichaeli, Curcuma zedoaria and Cinnamoum cassia, which were shown relatively potent anti-complementary activity were partially purified and analyzed their chemical properties. These activities were resistant to digestion with pronase but decreased by treatment with $NaIO_4$. These results may indicate that the complement activating ability in their herbs is due to polysaccharide. Furthermore, the anti-complementary activity of Areca catechu which was showed the most potent activity, was reduced partially in the absence of the $Ca^{++}\;ion$. After incubation of the normal human serum with partially purified polysaccharide of A. catechu in the absence of $Ca^{++}\;ion$, a cleavage of C3 in the serum was found to have occurred through immunoelectrophoresis using rabbit anti-human C3 serum. These results indicate that the mode of complement activation by polysaccharide of A. catechu is via both the alternative and classical pathway.

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Determination of Calibration Curve for Total Nitrogen Contents Analysis in Fresh Rice Leaves Using Visible and Near Infrared Spectroscopy (벼 생체엽신 질소함량 측정을 위한 근적외선분광분석의 검량식 작성)

  • Kwon Young-Rip;Baek Mi-Hwa;Choi Dong-Chil;Choi Joung-Sik;Choi Yeong-Geun
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.50 no.6
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    • pp.394-399
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    • 2005
  • Near Infrared Spectroscopy (NIRS) has been used as a tool for the rapid, accurate and nondestructive assay of the fresh rice leaf in nitrogen content. NIRS used in this study was visible and near infrared spectroscopy type instrument, Foss model 6500. To obtain a useful calibration equation, standard regression between the data was analyzed by chemical analysis and by NIRS method. Accuracy of calibration equation for nitrogen content on fresh leaf of rice were 0.879, 0.858 and 0.819, respectively. Accuracy of calibration equation after outlier treatment increased as 0.017, 0.02 and 0.061 improved each with 0.896, 0.878 and 0.880, respectively. Calibration equation combined using merge function after accuracy of calibration equation more increased by 0.911. Difference analysis value between calibration equation and lab value by kjeldahl showed $0.001\%$. With this as same result is the possibility of closing the deterioration of the sample in order to omit a construction and pulverization process it is judged with the fact that the nitrogen content measurement of the fresh rice leaf which the possibility of reducing an hour and an expense is by a near infrared spectroscopy technique will be possible.

When Dose Losses of Maternal Lymphocytes Response to Trophoblast Antigen or Alloantigen Occur in Women with a History of Recurrent Spontaneous Abortion? (반복유산을 경험한 환자에서 임신중 태반항원과 동종항원에 노출된 모체 림프구면역반응은 언제부터 소실되나?)

  • Choi, Bum-Chae;Hill, Joseph A.
    • Clinical and Experimental Reproductive Medicine
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    • v.25 no.2
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    • pp.115-122
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    • 1998
  • The maintenance of a viable pregnancy has long been viewed as an immunological paradox. The deveolping embryo and trophoblast are immunologically foreign to the maternal immune system due to their maternally inherited genes products and tissue-specific differentiation antigens (Hill & Anderson, 1988). Therefore, speculation has arisen that spontaneous abortion may be caused by impaired maternal immune tolerance to the semiallogenic conceptus (Hill, 1990). Loss of recall antigen has been reported in immunosuppressed transplant recipients and is associated with graft survival (Muluk et al., 1991; Schulik et al., 1994). Progesterone $(10^{-5}M)$ has immunosuppressive capabilities (Szekeres-Bartho et al., 1985). Previous study showed that fertile women, but not women with unexplained recurrent abortion (URA), lose their immune response to recall antigens when pregnant (Bermas & Hill, 1997). Therefore, we hypothesized that immunosuppressive doses of progesterone may affect proliferative response of lymphocytes to trophoblast antigen and alloantigen. Proliferative responses using $^3H$-thymidine ($^3H$-TdR) incorporation of peripheral blood mononuclear cells (PBMCs) to the irradiated allogeneic periperal blood mononuclear cells as alloantigen, trophoblast extract and Flu as recall antigen, and PHA as mitogen were serially checked in 9 women who had experienced unexplained recurrent miscarriage. Progesterone vaginal suppositories (100mg b.i.d; Utrogestan, Organon) beginning 3 days after ovulation were given to 9 women with unexplained RSA who had prior evidence of Th1 immunity to trophoblast. We checked proliferation responses to conception cycle before and after progesterone supplementation once a week through the first 7 weeks of pregnancy. All patients of alloantigen and PHA had a positive proliferation response that occmed in the baseline phase. But 4 out of 9 patients (44.4%) of trophoblast antigen and Flu antigen had a positive proliferative response. The suppression of proliferation response to each antigen were started after proliferative phase and during pregnancy cycles. Our data demonstrated that since in vivo progesterone treated PBMCs suppressed more T-lymphocyte activation and $^3H$-TdR incorporation compare to PBMCs, which are not influenced by progesterone. This data suggested that it might be influenced by immunosuppressive effect of progesterone. In conclusion, progesterone may play an important immunological role in regulating local immune response in the fetal-placental unit. Furthermore, in the 9 women given progesterone during a conception cycle, Only two (22%) repeat pregnancy losses occured in these 9 women despite loss of antigen responsiveness (one chemical pregnancy loss and one loss at 8 weeks of growth which was karyotyped as a Trisomy 4). These finding suggested that pregnancy loss due to fetal aneuploidy is not associated with immunological phenomena.

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Induction of Nrf2/ARE-mediated cytoprotective genes by red ginseng oil through ASK1-MKK4/7-JNK and p38 MAPK signaling pathways in HepG2 cells

  • Bak, Min Ji;Truong, Van-Long;Ko, Se-Yeon;Nguyen, Xuan Ngan Giang;Jun, Mira;Hong, Soon-Gi;Lee, Jong-Won;Jeong, Woo-Sik
    • Journal of Ginseng Research
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    • v.40 no.4
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    • pp.423-430
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    • 2016
  • Background: The induction of cellular defensive genes such as phase II detoxifying and antioxidant enzymes is a highly effective strategy for protection against carcinogenesis as well as slowing cancer development. Transcription factor Nrf2 (nuclear factor E2-related factor 2) is responsible for activation of phase II enzymes induced by natural chemopreventive compounds. Methods: Red ginseng oil (RGO) was extracted using a supercritical $CO_2$ extraction system and chemical profile of RGO was investigated by GC/MS. Effects of RGO on regulation of the Nrf2/antioxidant response element (ARE) pathway were determined by ARE-luciferase assay, western blotting, and confocal microscopy. Results: The predominant components of RGO were 9,12-octadecadienoic acid (31.48%), bicyclo[10.1.0] tridec-1-ene (22.54%), and 22,23-dihydrostigmasterol (16.90%). RGO treatment significantly increased nuclear translocation of Nrf2 as well as ARE reporter gene activity, leading to upregulation of heme oxygenase-1 and NAD(P)H:quinone oxidoreductase 1. Phosphorylation of the upstream kinases such as apoptosis signal-regulating kinase (ASK)1, mitogen-activated protein kinase (MAPK) kinase (MKK)4/7, c-Jun N-terminal kinase (JNK), and p38 MAPK were enhanced by treatment with RGO. In addition, RGO-mediated Nrf2 expression and nuclear translocation was attenuated by JNK inhibitor SP600125 and p38 MAPK inhibitor SB202190. Conclusion: RGO could be used as a potential chemopreventive agent, possibly by induction of Nrf2/ARE-mediated phase II enzymes via ASK1-MKK4/7-JNK and p38 MAPK signaling pathways.

GS28 Protects Neuronal Cell Death Induced by Hydrogen Peroxide under Glutathione-Depleted Condition

  • Lee, Hwa-Ok;Byun, Yu-Jeong;Cho, Kyung-Ok;Kim, Seong-Yun;Lee, Seong-Beom;Kim, Ho-Shik;Kwon, Oh-Joo;Jeong, Seong-Whan
    • The Korean Journal of Physiology and Pharmacology
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    • v.15 no.3
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    • pp.149-156
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    • 2011
  • Golgi SNAP receptor complex 1 (GS28) has been implicated in vesicular transport between intra-Golgi networks and between endoplasmic reticulum (ER) and Golgi. Additional role(s) of GS28 within cells have not been well characterized. We observed decreased expression of GS28 in rat ischemic hippocampus. In this study, we examined the role of GS28 and its molecular mechanisms in neuronal (SK-N-SH) cell death induced by hydrogen peroxide ($H_2O_2$). GS28 siRNA-transfected cells treated with $H_2O_2$ showed a significant increase in cytotoxicity under glutathione (GSH)-depleted conditions after pretreatment with buthionine sulfoximine, which corresponded to an increase of intracellular reactive oxygen species (ROS) in the cells. Pretreatment of GS28 siRNA-transfected cells with p38 chemical inhibitor significantly inhibited cytotoxicity; we also observed that p38 was activated in the cells by immunoblot analysis. We confirmed the role of p38 MAPK in cotransfected cells with GS28 siRNA and p38 siRNA in the cell viability assay, flow cytometry, and immunoblot. Involvement of apoptotic or autophagic processes in the cells was not shown in the cell viability, flow cytometry, and immunoblot analyses. However, pretreatment of the cells with necrostatin-1 completely inhibited $H_2O_2$-induced cytotoxicity, ROS generation, and p38 activation, indicating that the cell death is necroptotic. Collectively these data imply that $H_2O_2$ induces necroptotic cell death in the GS28 siRNA-transfected cells and that the necroptotic signals are mediated by sequential activations in RIP1/p38/ROS. Taken together, these results indicate that GS28 has a protective role in $H_2O_2$-induced necroptosis via inhibition of p38 MAPK in GSH-depleted neuronal cells.

Invitro and Virtual Screening of Bioactive Molecule from Mycelium of Trichoderma atroviride Inhibit the UDP-3-O-(R-3-hydroxymyristoyl)-N-acetylglucosamine Deacetylases (LpxC) for Treatment of Bacterial Infection

  • Saravanakumar, Kandasamy;Park, Cheol-Ho;Wang, Myeong-Hyeon
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.04a
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    • pp.67-67
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    • 2018
  • Trichoderma species are a rich source of metabolites, but less known for biomedical potential. This work deals with antibacterial and antioxidant potentials of intracellular non-cytotoxic metabolites, extracted from Trichoderma atroviride (KNUP001). A total of 53 fractions was collected by column chromatography and tested for cytotoxicity by MTT assay. Only one fraction (F41) was found to be non-toxic to Vero cells with $95.4{\pm}0.61%$ of survival. The F41 was then subjected to chemical analysis, antibacterial and antioxidant assays. The F41 at $500{\mu}g.ml^{-1}$ showed the total antioxidant of $48.70{\pm}2.90%$, DPPH radical scavenging activity of $37.25{\pm}2.25$, nitric oxide (NO) radical scavenging activity of $54.55{\pm}1.95$ and $H_2O_2$ radical scavenging activity of $43.75{\pm}3.21$. The F41 at $25{\mu}g.ml^{-1}$ displayed antibacterial activity against E. coli ($14.25{\pm}0.2mm$), P. mirabilis ($10.4{\pm}0.6mm$), S. dysenteriae ($18.6{\pm}03mm$), S. paratyphi A ($14.1{\pm}1.1mm$), E. aerogenes ($5.6{\pm}0.4mm$) and S. marcescens ($14.25{\pm}0.2mm$). GC-MS analysis revealed the dominant presence of oleic acid C 18.1 (63.18%), n-hexadecanoic acid (6.17%), and ethyl oleate (4.93%) and potent molecules such as 8-[(2E)-2-(3-hydroxybenzylidene)hydrazinyl]-1,3,7-trimethyl-3,7-dihydro-1H-purine-2,6-dione, 2-(Dimethylamino)ethyl (1Z)-N-hydroxy-2-(4-morpholinyl)-2-oxoethanimidothioate, Fluorene in the F41, and virtual study revealed that these molecules are likely responsible for the antibacterial activities of F41. Hence, further investigation deserves on purification and characterization of the active metabolites from T. atroviride strain KNUP001 towards developing molecular leads to effective antibacterial drugs, and non-toxic to host cells.

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A STUDY ON THE RESISTANCE OF WEAR AND CYTOTOXICITY OF THE TITANIUM SURFACE AFTER FILM DEPOSITIONS (박막증착시 티타늄 표면의 마손저항도와 세포독성에 관한 연구)

  • Kim Hyung-Woo;Kim Chang-Whe;Kim Yung-Soo
    • The Journal of Korean Academy of Prosthodontics
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    • v.39 no.1
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    • pp.84-95
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    • 2001
  • Titanium is widely used in dentistry for its low density, high strength, fatigue resistance, corrosion resistance, and biocompatibility. But it has a tendency of surface damage under circumstance of friction and impact for its low hardness of the surface. Coating is one of methods fir increasing surface hardness. Its effect is to improve surface physical characteristics without change of titanium. Diamond-like carbon and titanium nitride are known for its high hardness of the surface. So that this study was aimed at the wear test and the cytotoxicity test of the commercially pure titanium and Ti-6Al-4V alloy which were deposited by diamond-like carbon film or titanium nitride film to acertain improvement of the surface hardness and the biocompatibility. A disk (25mm diameter, 2mm thickness) was made of commercially pure titanium and Ti-6Al-4V alloy and these substrates were deposited by diamond-like carbon film or titanium nitride film. Diamond-like carbon film was deposited by the method of radiofrequency plasma assisted chemical vapor deposition and titanium nitride film was deposited by the method of reactive arc ion plating. Then these substrates were tested about wear characteristics by the pin-on-disk type wear tester in which ruby ball was used as a wear causer under the load of 32N, The fracture cycles were measured by rotating the substrates until their films were fractured. The wear volume was measured after 150 cycles and 3,000 cycles using surface profiler. The cytotoxicity test was peformed by the method of the MTT assay. The results were as follows : 1. In the results of the wear volume test, commercially pure titanium and titanium alloy which were coated by diamond-like carbon film or titanium nitride aim had higher resistance against wear than the substrates which were not coated by any films (P<0.05). 2. In the results of the fracture cycle test and the wear volume test, diamond-like carbon film had higher resistance against wear than titanium nitride film (P<0.05). 3. In both coatings of diamond-like carbon aim and titanium nitride film, Ti-6Al-4V alloy had higher resistance against wear than commercially pure titanium (P<0.05) 4. In the results of the cytotoxicity test, diamond-like carbon film and titanium nitride film had little cytotoxicity as like commercially pure titanium or Ti-6Al-4V alloy (P>0.05).

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Comparative study on the physicochemical properties and cytocompatibility of microporous biphasic calcium phosphate ceramics as a bone graft substitute (미세다공성 Biphasic calcium phosphate ceramics의 골이식 대체재로서의 기본특성에 대한 비교연구)

  • Park, Kwang-Bum;Park, Jin-Woo;Ahn, Hyun-Uk;Yang, Dong-Jun;Choi, Seok-Kyu;Jang, II-Sung;Yeo, Shin-Il;Suh, Jo-Young
    • Journal of Periodontal and Implant Science
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    • v.36 no.4
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    • pp.797-808
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    • 2006
  • Objective : The purpose of this study was to evaluate the physicochemical properties and cytocompatibility of microporous, spherical biphasic calcium phosphate(BCP) ceramics with a 60/40 $hydroxyapatite/{\beta}$ -tricalcium phosphate weight ratio for application as a bone graft substitute. Materials and Methods : Microporous, spherical BCP granules(MGSB) were prepared and their basic characteristics were compared with commercially available BCP(MBCP; Biomatlante, France) and deproteinized bovine bone mineral(Bio-Oss; GBistlich-Pharma, Switzerland, BBP; Oscotec. Korea), Their physicochemical properties were evaluated by scanning electron microscopy, X-ray diffractometry, Fourier-transform infrared spectroscopy, inductively coupled plasma atomic emission spectrometer, and Brunauer-Emmett-Teller method. Cell viability and proliferation of MC3T3-El cells on different graft materials were evaluated. Results : MGSB granules showed a chemical composition and crystallinity similar with those in MBCP, they showed surface structure characteristic of three dimensionally, well-interconnected micropores. The results of MTT assay showed increases in cell viablity with increasing incubation times. At 4d of incubation, MGSB, MBCP and BBP showed similar values in optical density, but Bio-Oss exhibited significantly lower optical density compared to other bone substitutes(p <0,05). MGSB showed significantly greater cell number compared to other bone substitutes at 3, 5, and 7d of incubation(p <0,05), which were similar with those in polystyrene culture plates. Conclusion: These results indicated the suitable physicochemical properties of MGSB granules for application as an effective bone graft substitute. which provided compatible environment for osteoblast cell growth. However, further detailed studies are needed to confirm its biological effects on bone formation in vivo.