• Title/Summary/Keyword: bio assay

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Possibility of Undifferentiated Human Thigh Adipose Stem Cells Differentiating into Functional Hepatocytes

  • Lee, Jong Hoon;Lee, Kuk Han;Kim, Min Ho;Kim, Jun Pyo;Lee, Seung Jae;Yoon, Jinah
    • Archives of Plastic Surgery
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    • v.39 no.6
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    • pp.593-599
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    • 2012
  • Background This study aimed to investigate the possibility of isolating mesenchymal stem cells (MSCs) from human thigh adipose tissue and the ability of human thigh adipose stem cells (HTASCs) to differentiate into hepatocytes. Methods The adipose-derived stem cells (ADSCs) were isolated from thigh adipose tissue. Growth factors, cytokines, and hormones were added to the collagen coated dishes to induce the undifferentiated HTASCs to differentiate into hepatocyte-like cells. To confirm the experimental results, the expression of hepatocyte-specific markers on undifferentiated and differentiated HTASCs was analyzed using reverse transcription polymerase chain reaction and immunocytochemical staining. Differentiation efficiency was evaluated using functional tests such as periodic acid schiff (PAS) staining and detection of the albumin secretion level using enzyme-linked immunosorbent assay (ELISA). Results The majority of the undifferentiated HTASCs were changed into a more polygonal shape showing tight interactions between the cells. The differentiated HTASCs up-regulated mRNA of hepatocyte markers. Immunocytochemical analysis showed that they were intensely stained with anti-albumin antibody compared with undifferentiated HTASCs. PAS staining showed that HTASCs submitted to the hepatocyte differentiation protocol were able to more specifically store glycogen than undifferentiated HTASCs, displaying a purple color in the cytoplasm of the differentiated HTASCs. ELISA analyses showed that differentiated HTASCs could secrete albumin, which is one of the hepatocyte markers. Conclusions MSCs were islolated from human thigh adipose tissue differentiate to heapatocytes. The source of ADSCs is not only abundant abdominal adipose tissue, but also thigh adipose tissue for cell therapy in liver regeneration and tissue regeneration.

Biological activities of Brassica rapa (Turnip) callus extracts by plant cell culture technology (식물세포배양기술을 이용한 순무 캘러스 추출물의 생리활성)

  • Shin, Su Young;Moh, Sang Hyun;Hwang, You Jin
    • Journal of Plant Biotechnology
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    • v.43 no.2
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    • pp.248-254
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    • 2016
  • The purpose of this study was to investigate biological activities of Brassica rapa (Turnip) plant callus extracts of Ganghwa-gun of Incheon city using water, ultrasonic wave and ethanol extractions to develop functional materials. DPPH radical scavenging activities of the callus extracts were increased in a concentration-dependent manner, as compared with control. The astringent effects of the ethanol extracts were higher, as compared to water and ultrasonic extracts. In the collagen synthesis assay, the ethanol extract showed significant anti-wrinkle effects of 59% and 78% at a concentration of 5 ppm and 10 ppm, respectively. These results suggested that water, ultrasonic wave and ethanol extracts of turnip plant calluses are natural antioxidant sources. Especially, the ethanol extract can be regarded as a functional, natural cosmetic material with astringent and anti-wrinkle effects.

The Utility of Measuring Assimiliable Organic Carbon (AOC) as an Indicator of Biostability in Distribution Systems for Finished Water

  • Chang, Young-Cheol;Toyama, Tadashi;Jung, Kweon;Kikuchi, Shitaro
    • Journal of Environmental Health Sciences
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    • v.32 no.6
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    • pp.539-542
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    • 2006
  • The objective of this paper is to compare the applicability of assimilable organic carbon (AOC) or biodegradable dissolved organic carbon (BDOC) for quantifying biodegradable organic material (BOM) and bio-stability in distribution systems for a variety of finished waters. The study the data is derived from was part of an AWWARF and Tampa Bay Water tailored collaboration project to determine the effect of blending different waters on distribution system water quality. Seven different finished waters were produced from surface, ground, or brackish water on site and fed 18 independent pilot distribution systems (PDSs), either as single finished water or as a blend of several finished waters. AOC and BDOC have often been used as indicators of bacterial regrowth potential in distribution systems. In this study, AOC was the more useful assay of the two for the BOM concentrations observed in the PDSs. BDOC did not distinguish BOM while AOC did at the low BOM levels from many of the advanced treatments (e.g. RO, $O^3/BAC$). AOC in contrast allowed much more meaningful calculations of the consumption or production of AOC as the blends passed through the PDSs even for very low BOM blends. In addition, meaningful trends corresponding to changes in heterophic plate count (HPC) were observed for AOC but not for BDOC. Moreover, AOC stability was associated with waters produced from advanced membrane treatment.

Effects of various receptor antagonists on the peripheral antinociceptive activity of aqueous extracts of Dicranopteris linearis, Melastoma malabathricum and Bauhinia purpurea leaves in mice

  • Zakaria, Zainul Amiruddin;Sodri, Nurul Husna;Hassan, Halmy;Anuar, Khairiyah;Abdullah, Fatimah Corazon
    • CELLMED
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    • v.2 no.4
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    • pp.38.1-38.6
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    • 2012
  • The present study aimed to determine the possible mechanisms of the peripheral antinociception of the aqueous extracts of Dicranopteris linearis (AEDL), Melastoma malabathricum (AEMM) and Bauhinia purpurea (AEBP) leaves in mice. Briefly, the antinociceptive profile of each extract (300, 500, and 1000 mg/kg; subcutaneous (s.c.)), was established using the abdominal constriction test. A single dose (500 mg/kg) of each extract (s.c.) was pre-challenged for 10 min with various pain receptors' antagonists or pain mediators' blockers and 30 min later subjected to the antinociceptive assay to determine the possible mechanism(s) involved. Based on the results obtained, all extracts exerted significant (p < 0.05) antinociceptive activity with dose-dependent activity observed only with the AEMM. Furthermore, the antinociception of AEDL was attenuated by naloxone, atropine, yohimbine and theophylline; AEMM was reversed by yohimbine, theophylline, thioperamide, pindolol, reserpine, and 4-chloro-DL-phenylalanine methyl ester hydrochloride; and of AEBP was inhibited by naloxone, haloperidol, yohimbine and reserpine. In conclusion, the antinociceptive activity of those extracts possibly involved the activation of several pain receptors (i.e. opioids, muscarinic, ${\alpha}_2$-adrenergic and adenosine receptors, adenosine, H3-histaminergic and $5HT_{1A}$, dopaminergic receptors).

Anti-inflammatory and Anti-oxidative Effects of Alpiniae Oxyphyllae Fructus Hot Aqueous Extract in Lipopolysaccharide (LPS)-stimulated Macrophages

  • Jo, Na Young;Lee, Pyeong Jae;Roh, Jeong Du
    • Journal of Acupuncture Research
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    • v.34 no.2
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    • pp.83-91
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    • 2017
  • Objectives : Alpiniae oxyphyllae Fructus (AOF) is an herbal medicine, which has been used for the treatment of fatigue, chills, and poor physical conditions. The objective of this study was to investigate the anti-inflammatory and anti-oxidative effects of AOF hot aqueous extract. Methods : The cytotoxicity of AOF extract was evaluated using the MTT assay. Nitric oxide (NO) production was measured by the Griess reaction. Prostaglandin $E_2$ ($PGE_2$) production was measured by a commercial competitive enzyme immunoassay. Cytokine production (IL-1tion co6, and TNF- F- was measured by ELISA. The anti-oxidative effect of AOF extracts was measured by the DPPH method. Polyphenol and flavonoid contents were measured by Folin-Ciocalteu's phenol reagent and aluminum chloride, respectively. Results : AOF hot aqueous extract did not show toxicity at doses of 25, 50, 100, and $200{\mu}g/mL$. AOF extract significantly inhibited NO production at doses of 100 and $200{\mu}g/mL.PGE_2$ production was inhibited by AOF extract treatment at doses of 100 and $200{\mu}g/mL$. AOF extracts reduced IL-6 production in a dose-dependent manner. IL-1ent maTNF- F- 1ent mannerd IL-6 production in uction at doses of 100 and ${\mu}g/mL$. The DPPH free radical scavenging capability was above 50% at $200{\mu}g/mL$. Conclusion : This study suggests that AOF hot aqueous extract may exert anti-inflammatory and anti-oxidative effects in a dose-dependent manner. Further studies are required for validating the safety and efficacy of AOF.

Chemical Composition and Biological Activities of Essential Oils Extracted from Korean Endemic Citrus Species

  • Baik, Jong-Seok;Kim, Sang-Suk;Lee, Jung-A;Oh, Tae-Heon;Kim, Ji-Young;Lee, Nam-Ho;Hyun, Chang-Gu
    • Journal of Microbiology and Biotechnology
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    • v.18 no.1
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    • pp.74-79
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    • 2008
  • The aim of this study was to analyze the chemical composition of 14 kinds of citrus oils and to test their biological activities. Citrus essential oils were obtained by steam distillation from immature fruits collected from Jeju Island and were analyzed using gas chromatograph (GC)-flame ionization detectors (FID) and GC-MS. Limonene (55.4% to 91.7%), myrcene (2.1% to 32.1%), ${\alpha}$-pinene (0.6% to 1.6%) and linalool (0.4% to 6.9%) were the major components in most citrus species. To evaluate in vitro antibacterial activity, all essential oils were tested against Propionibacterium acnes and Staphylococcus epidermidis. Nine out of fourteen citrus oils exhibited antibacterial activity against P. acnes, but not against S. epidermidis. The effects of the citrus oils on DPPH radical scavenging, superoxide radical anion scavenging, nitric oxide radical, and cytotoxicity were also assessed. Three essential citrus oils, Joadeung, Dongjunggyul, and Bujiwha, exhibited potent inhibitory effects on nitric oxide production. Two essential oils, Dongjunggyul and Joadeung, showed potent free radical scavenging activities in the DPPH assay. For future applications in cosmetic products, we also performed MTT assays in a human dermal fibroblast cell line. The majority of the essential oils showed no cytotoxicity. The results indicate that citrus essential oils can be useful natural agents for cosmetic application.

Inhibitory effect of Alzheimer's in APP-induced Neuro 2A cells by JangWonHwanGagambang(JWHG) water extract (장원환가감방 수추출물(水抽出物)이 아밀로이드 전구단백질으로 유도된 생쥐의 신경아세포주에서의 항치매 효과)

  • Kim, Sang-Tae;Lee, Jong-Hwa;Kim, Tae-Hyeon;Son, Hyeong-Jin;Han, Pyeong-Leem;Kang, Hyung-Won
    • Journal of Oriental Neuropsychiatry
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    • v.18 no.2
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    • pp.13-24
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    • 2007
  • Objective : Recent studies indicate that the deposition of ${\beta}-amyloid$ ($A{\beta}$) is related in the pathogenesis of Alzheimer's disease (AD), but the underlying mechanism is still not clear. Method : To investigate the potential cellular functions of APP and water extract of the JangwonHwangagambang (JWHG), we use as in vitro model, neuro 2A cells were treated with either JWHG or its oriental medicines, and the effect in APP expression was determined by MTT and LDH assay. JWHG have been shown to be neuroprotective in different model systems. We asked whether JWHG treatment would influence cell survival and AD-like pathology in APP-induced neuronal cells. Result : JWHG and water extracts of some oriental medicine has attenuated high cell death in vitro. JWHG-treated cells increased percentage of cell survival more longly than controls. JWHG had significantly increas neurite outgrowth in the as compared to control cells. Conclusion : These results suggest that JWHG prevent APP-induced neurotoxicity through attenuating oxidative stress, and may be useful as potential therapeutic agents for AD.

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Effects of White Radish (Raphanus sativus) Enzyme Extract on Hepatotoxicity

  • Lee, Sang-Wha;Yang, Kwang-Mo;Kim, Jung-Ki;Nam, Byung-Hyouk;Lee, Chang-Min;Jeong, Min-Ho;Seo, Su-Yeong;Kim, Gi-Yong;Jo, Wol-Soon
    • Toxicological Research
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    • v.28 no.3
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    • pp.165-172
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    • 2012
  • Raphanus sativus (Cruciferaceae), commonly known as radish is widely available throughout the world. From antiquity it has been used in folk medicine as a natural drug against many toxicants. The present study was designed to evaluate the hepatoprotective activity of radish (Raphanus sativus) enzyme extract (REE) in vitro and in vivo test. The $IC_{50}$ values of REE in human liver derived HepG2 cells was over 5,000 ${\mu}g/ml$ in tested maximum concentration. The effect of REE to protect tacrine-induced cytotoxicity in HepG2 cells was evaluated by MTT assay. REE showed their hepatoprotective activities on tacrine-induced cytotoxicity and the $EC_{50}$ value was 1,250 ${\mu}g/ml$. Silymarin, an antihepatotoxic agent used as a positive control exhibited 59.7% hepatoprotective activity at 100 ${\mu}g/ml$. Moreover, we tested the effect of REE on carbon tetrachloride ($CCl_4$)-induced liver toxicity in rats. REE at dose of 50 and 100 mg/kg and silymarin at dose of 50 mg/kg were orally administered to $CCl_4$-treated rats. The results showed that REE and silymarin significantly reduced the elevated levels of serum enzyme markers induced by $CCl_4$. The biochemical data were supported by evaluation with liver histopathology. These findings suggest that REE, can significantly diminish hepatic damage by toxic agent such as tacrine or $CCl_4$.

Contents of Isoflavones and Antioxidative Related Compounds in Soybean Leaf, Soybean Leaf Jangachi, and Soybean Leaf Kimchi (콩잎 밑 콩잎 요리의 이소플라본 함량 및 항산화 관련 성분들의 비교)

  • Ryu Seung-Hee;Lee Hye-Suk;Lee Young-Soon;Moon Gap-Soon
    • Korean journal of food and cookery science
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    • v.21 no.4 s.88
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    • pp.433-439
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    • 2005
  • Soybean is an important plant as it is the source of protein and oil as well as various phytochemicals that are related with biological activity. Over the past decades, scientists have conducted considerable research on the physiological properties of soybeans, especially isoflavones, which are the characteristic components in soybeans. However, there is no research on the properties or the bio-functionality of soybean leaf. Jangachi and kimchi are two of the traditional special dishes of Gyungsang Province in Korea which we made from soybean leaves. Depending on the recipe, green or yellow soybean leaves are used for the preparation of these two side dishes. We compared the antioxidative activity and measured the contents of isoflavones, total phenol, chlorophylls, carotenoids, and vitamin C in the ingredients (green and yellow soybean leaf) and the final side dishes (jangachi and kimchi). We Int report that isoflavones were contained in soybean leaf and that jangachi had the highest isoflavone contents among the samples. Yellow soybean leaf contained higher isoflavones than green soybean lear and kimchi. From the TEAC assay results, the sequence or antioxidative activities was yellow soybean leaf > soybean leaf jangachi > green soybean leaf > soybean leaf kimchi. The sequence was the same with total phenol contents, indicating that antioxidative activity is highly related with total phenol level. Chlorophylls, carotenoids and vitamin C existed abundantly in green soybean leaf. In conclusion, soybean leaf could be a good material for health due to the presence of isoflavones and the other useful antioxidants mentioned above.

Purification and Characterization of Anticoagulant Protein from the Tabanus, Tabanus bivittatus

  • Ahn Mi-Young;Hahn Bum-Soo;Lee Pyeong-Jae;Wu Song-Ji;Kim Yeong-Shik
    • Archives of Pharmacal Research
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    • v.29 no.5
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    • pp.418-423
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    • 2006
  • Tabanus anticoagulant protein (TAP) was isolated from the whole body of the tabanus, Tabanus bivittatus, using three purification steps (ammonium sulfate fractionation, gel filtration on Bio-Gel P-60, and ion exchange chromatography on DEAE Sephadex gel). The purified TAP, with a molecular weight of 65 kDa, was assessed to be homogeneous by SDS-polyacrylamide gel electrophoresis, and an isoelectric point of 7.9 was determined by isoelectric focusing. The internal amino acid sequence of the purified protein was composed of Ser-Leu-Asn-Asn-Gln-Phe-Ala-Ser-Phe-lle-Asp-Lys-Val-Arg. The protein was activated by $Cu^{2+}\;and\;Zn^{2+}$, and the optimal conditions were found to be at pH $3\sim6\;and\;40\sim70^{\circ}C$. Standard coagulation screen assays were used to determine thrombin time and activated partial thromboplastin time. Chromogenic substrate assays were performed for thrombin and factor Xa activity. TAP considerably prolonged human plasma clotting time, especially activated partial thromboplastin time in a dose-dependent manner; it showed potent and specific antithrombin activity in the chromogenic substrate assay. Specific anti-factor Xa activity in TAP was not detected. Overall, this result suggested that TAP has significant anticoagulant activity on blood coagulation system.