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One-Step Simultaneous Immunochromatographic Strip Test for Multianalysis of Ochratoxin A and Zearalenone

  • Shim, Won-Bo;Dzantiev, Boris B.;Eremin, Sergei A.;Chung, Duck-Hwa
    • Journal of Microbiology and Biotechnology
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    • v.19 no.1
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    • pp.83-92
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    • 2009
  • Individual immunochromatographic assays (ICG) for ochratoxin A (OTA) and zearalenone (ZEA) were optimized and used in the development of a one-step simultaneous immunochromatographic assay (OS-ICG) for the rapid multianalysis of two mycotoxins in corn samples. The nitrocellulose membrane of the OS-ICG was treated with OTA-bovine serum albumin (BSA), ZEA-ovalbumin (OVA), and anti-mouse IgG in the OTA test, ZEA test, and control zones, respectively. Monoclonal antibody-gold conjugates (OTA3 MAb-gold and ZEA2C5 MAb-gold) were sprayed onto the conjugate pad. The visual detection limits were 2.5 and 5 ng/ml for OTA and ZEA, respectively, and the results were obtained within 15 min after starting the analysis. An efficient, simple, and rapid extraction method using 30% MeOH/PBS was established and validated by analyzing the corn samples spiked with OTA/ZEA mixtures (0/0, 5/10, 10/20, and $20/30\;{\mu}g/kg$). The cut-off values of the OS-ICG for the spiked corn were 5 and $10\;{\mu}g/kg$ for OTA and ZEA, respectively. Natural corn samples were analyzed by OS-ICG, direct competitive enzyme-linked immunosorbent assay (DC-ELISA), and HPLC. Results of the OS-ICG were in good agreement with those obtained by DC-ELISA and HPLC. The developed OS-ICG offers a rapid, easy-to-use, and portable analytical system and can be used as a convenient qualitative tool for the on-site simultaneous determination of OTA and ZEA in cereals, food, and agricultural products in one analytical cycle.

The Effects of Cereal Supplementation on the Iron Nutritional Status in the Korean Institutionalized Children (사회복지시설 아동에게 시리얼보충이 철분영양상태에 미치는 영향)

  • Chung, Hae-Kyung;Kang, Myung-Hwa
    • Journal of the Korean Society of Food Culture
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    • v.18 no.6
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    • pp.575-583
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    • 2003
  • To evaluate the effect of cereal supplementation on children's iron nutritional status of Korean institutionalized was designed. Dietary survey was carried out methods of food weighting in the breakfast or/and dinner, and record interview in lunch (n=74). A nutritional intervention study was carried out through supplementing cereal for 4 weeks in 24 children of 1 institution from 4 to 12 years. The children received 3.6mg elemental Fe(as 100g cereal) per day. Blood samples were drawn before and after supplementation. Nutrients which children's intake was less than two-thirds of Korean RDA were Vit A, Vit B1, Vit B2, Ca and Fe. The mean daily intakes of iron were 5.1mg for male and 4.9mg for female and 52.3% for male and 45.4% for female of Korean RDA. The proportions of children with iron depletion assessed by TIBC(>360mg/dl) and serum ferritin(<20ng/ml) were 56.6% and 58.7%, respectively. The proportions of children with the iron deficient erythropoiesis assessed by serum iron(<70ml/dl), Hb(<12g/dl), and Hct(<36%) were 76.0%, 58.7%, and 64.0%, respectively. After cereal supplementation, in anemic children, levels of Hct(p<0.001), serum iron(p<0.001) and transferrin saturation(p<0.001) were significantly increased. The effect of cereal supplementation in children with iron deficient erythropoeisis was more effective to improve the iron nutritional status than children with iron depletion. It was concluded that cereal supplementation program in anemic children was also effective to improve iron nutritional status.

Effect of lipoaspirate cell autograft on proliferation and collagen synthesis of diabetic fibroblasts in vitro (지방기질세포 치료가 당뇨섬유아세포의 증식과 교원질합성에 미치는 영향)

  • Song, Sun Ho;Han, Seung Kyu;Chun, Kyung Wook;Kim, Woo Kyung
    • Archives of Plastic Surgery
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    • v.36 no.6
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    • pp.679-684
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    • 2009
  • Purpose: Human lipoaspirate cells are relatively easy to obtain in large quantities without cell culture. The aim of this in vitro pilot study was to determine the effect of cell therapy using uncultured lipoaspirate cells on cell proliferation and collagen synthesis of diabetic fibroblasts, which are the major contributing factors in wound healing. Methods: In order to get diabetic fibroblasts, dermis tissues were obtained from foot skin of diabetic patients who underwent debridements or toe amputations(n = 4). In order to isolate lipoaspirate cells, the same diabetic patients' abdominal adipose tissues were obtained by liposuction. The diabetic fibroblasts were co - cultured with or without autogenous lipoaspirate cells using porous culture plate insert. Initial numbers of the lipoaspirate cells and diabetic fibroblasts seeded were 15,000 cells/well, respectively. For cell proliferation assay, two treatment groups were included. In group I, diabetic fibroblasts were cultured with the insert having no cells, which serves as a control. In group II, the lipoaspirate cells were added in the culture plate insert. For collagen synthesis assay, one additional group(group III), in which diabetic fibroblasts were not seeded in the well and only lipoaspirate cells inside the insert were incubated without diabetic fibroblasts, was included for a reference. Results: One hundred to one hundred sixty thousand lipoaspirate cells were isolated per ml of aspirated adipose tissue. After 3 - day incubation, the mean cell numbers in group I and II were 17,294/well and 22,163/well. The mean collagen level in group I, II, and III were 29, 41, and 2 ng/ml, respectively. These results imply that both cell proliferation and collagen synthesis in the lipoaspirate cell treatment group were 28 and 44 percents higher than in the control group, respectively(p < 0.05). Conclusion: Uncultured lipoaspirate cell autografts may stimulate the wound healing activity of diabetic fibroblasts.

Vitamin D status and its association with cardiometabolic risk factors in Korean adults based on a 2008-2010 Korean National Health and Nutrition Examination Survey

  • Chung, Ji-Youn;Hong, Sung-Ho
    • Nutrition Research and Practice
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    • v.7 no.6
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    • pp.495-502
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    • 2013
  • Recent studies suggest that vitamin D deficiency and cardiometabolic disorders are becoming increasingly more prevalent across multiple populations. However, there is a lack of comprehensive data for Korean adults. We investigated the vitamin D status, the prevalence of vitamin D deficiency and its association with metabolic syndrome (MS) risk in Korean adults aged 20 years or older. The study subjects (n = 18,305) were individuals who participated in the Korean National Health Examination and Nutrition Survey (KNHANES) in 2008-2010. Vitamin D status (25-hydroxyvitamin D [25(OH)D]) was categorized as < 20, 21-29, and ${\geq}$ 30 ng/mL, which are the cut-off points for deficiency, insufficiency and normal limits. A wide variety of cardiometabolic risk factors were compared according to the vitamin D status. Vitamin D deficiency was found in 53.9% of men and 70.5% of women. Mean BMI, systolic BP, HbA1c and low density lipoprotein cholesterol (LDL-C) were highest in the vitamin D deficiency group in both genders. Further, the MS was most prevalent in the vitamin D deficiency group in both genders (12.3%, P = 0.002 in men and 9.2%, P < 0.001 in women). Compared to the vitamin D normal group, the adjusted odds ratio (ORs) (95% confidence interval [95% CI]) for MS in the vitamin D deficiency group were 1.46 (1.05-2.02) in men and 1.60 (1.21-2.11) in women, after adjusting for confounding variables. In conclusion, Vitamin D deficiency is a very common health problem in Korean adults and is independently associated with the increasing risk of MS.

Anti-inflammatory Effects of 8α-hydroxy pinoresinol isolated from Nardostachys jatamansi on Lipopolysaccharide-induced Inflammatory Response in RAW 264.7 Cells. (LPS로 유도된 RAW 264.7 세포의 염증반응에서 감송향(甘松香)에서 추출한 8α-hydroxy pinoresinol의 항염증 효과)

  • Choi, Sun Bok;Park, Sung-Joo
    • The Korea Journal of Herbology
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    • v.31 no.5
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    • pp.1-6
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    • 2016
  • Objectives : Nardostachys jatamansi (NJ) is a medicinal herb that has been reported in various traditional systems of medicine for its use in antispasmodic, a digestive stimulant, skin diseases. Previous studies have already reported that NJ effectively protects against inflammation. However, the active compound in NJ is unknown. Therefore, in the present study, we analyzed effects of a compound, 8α-hydroxy pinoresinol (HP), isolated from NJ against lipopolysaccharide (LPS) induced inflammation in RAW 264.7 cells.Methods : To examine the anti-inflammatory effect of HP against LPS, intraperitoneally pre-treat the HP (100, 200, 500 and 1,000 nM) 1 h prior to LPS challenges. LPS was stimulated with 500 ng/ml in RAW 264.7 cells. To identify the anti-inflammatory effect of HP, we measured inflammatory mediators such as inducible nitric oxide synthase (iNOS) and its derivative nitric oxide (NO), cyclooxygenase-2 (COX-2), prostaglandin E2 (PGE2). Also we evaluated molecular mechanisms including mitogen-activated protein kinases (MAPKs) and nuclear factor-kappaB (NF-κB) activation by western blot.Results : The HP inhibited production of inflammatory mediators, such as iNOS and its derivative NO, COX-2 and PGE2 in LPS- induced inflammationin RAW 264.7 cells. Additionally, HP also inhibited activation of p38 pathway signaling but not extracellularsignal-regulatedkinase (ERK), c-jun NH2-terminal kinase (JNK), and NF-κB.Conclusion : Our results suggest that HP has anti-inflammatory functions through the dephosphorylation of p38 and HP can provide beneficial strategy for prevention and therapy of inflammation.

Study on the Inclusion Behavior of Sulfobutylether-β-Cyclodextrin with Perphenazine by Flow Injection Chemiluminescence

  • Shen, Minxia;Lv, Hairu;Song, Zhenghua
    • Bulletin of the Korean Chemical Society
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    • v.34 no.11
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    • pp.3199-3205
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    • 2013
  • The inclusion behavior of sulfobutylether-${\beta}$-cyclodextrin (SBE-${\beta}$-CD) with perphenazine (PPH) was first studied by flow injection (FI)-chemiluminescence (CL) analysis with proposed $lg[(I_0-I_s)/I_s]=lgK_{P-CD}+nlg[C_{PPH}]$ model and molecular docking. Results showed that a 1:1 complex of SBE-${\beta}$-CD/PPH could online form, with the formation constant $K_{P-CD}$ of $2.57{\times}10^7Lmol^{-1}$ at 298 K. The thermodynamic parameters showed that the inclusion behavior of SBE-${\beta}$-CD/PPH was a spontaneous process by hydrophobic interaction. The molecular docking results revealed PPH entered into the larger cavity of SBE-${\beta}$-CD with two hydrogen bonds. Based on the linear relationship of the decrement of luminol/SBE-${\beta}$-CD/PPH CL intensity against the logarithm of PPH concentration ranging from 0.03 to 30.0 ng $mL^{-1}$, the present FI-CL analysis using luminol/SBE-${\beta}$-CD/PPH system was successfully applied to PPH determination in biological fluids and tablets with recoveries from 94.5 to 105.6% and RSDs less than 2.6% (n = 5).

Effects of Lubiprostone on Pacemaker Activity of Interstitial Cells of Cajal from the Mouse Colon

  • Jiao, Han-Yi;Kim, Dong Hyun;Ki, Jung Suk;Ryu, Kwon Ho;Choi, Seok;Jun, Jae Yeoul
    • The Korean Journal of Physiology and Pharmacology
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    • v.18 no.4
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    • pp.341-346
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    • 2014
  • Lubiprostone is a chloride ($Cl^-$) channel activator derived from prostaglandin $E_1$ and used for managing constipation. In addition, lubiprostone affects the activity of gastrointestinal smooth muscles. Interstitial cells of Cajal (ICCs) are pacemaker cells that generate slow-wave activity in smooth muscles. We studied the effects of lubiprostone on the pacemaker potentials of colonic ICCs. We used the whole-cell patch-clamp technique to determine the pacemaker activity in cultured colonic ICCs obtained from mice. Lubiprostone hyperpolarized the membrane and inhibited the generation of pacemaker potentials. Prostanoid $EP_1$, $EP_2$, $EP_3$, and $EP_4$ antagonists (SC-19220, PF-04418948, 6-methoxypyridine-2-boronc acid N-phenyldiethanolamine ester, and GW627368, respectively) did not block the response to lubiprostone. L-NG-nitroarginine methyl ester (L-NAME, an inhibitor of nitric oxide synthase) and 1H-[1,2,4]oxadiazolo[4,3,-a]quinoxalin-1-one (ODQ, an inhibitor of guanylate cyclase) did not block the response to lubiprostone. In addition, tetraethylammonium (TEA, a voltage-dependent potassium [$K^+$] channel blocker) and apamin (a calcium [$Ca^{2+}$]-dependent $K^+$ channel blocker) did not block the response to lubiprostone. However, glibenclamide (an ATP-sensitive $K^+$ channel blocker) blocked the response to lubiprostone. Similar to lubiprostone, pinacidil (an opener of ATP-sensitive $K^+$ channel) hyperpolarized the membrane and inhibited the generation of pacemaker potentials, and these effects were inhibited by glibenclamide. These results suggest that lubiprostone can modulate the pacemaker potentials of colonic ICCs via activation of ATP-sensitive $K^+$ channel through a prostanoid EP receptor-independent mechanism.

A Novel Metalloprotease from the Wild Basidiomycete Mushroom Lepista nuda

  • Wu, Y.Y.;Wang, H.X.;Ng, T.B.
    • Journal of Microbiology and Biotechnology
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    • v.21 no.3
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    • pp.256-262
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    • 2011
  • A 20.9-kDa metalloprotease was isolated from dried fruiting bodies of the wild basidiomycete mushroom Lepista nuda. The N-terminal amino acid sequence of the protease was seen to be ATFVLTAATNTLFTA, thus displaying no similarity with the sequences of previously reported metalloproteases. The protease was purified using a procedure that entailed ion-exchange chromatography on CM-Cellulose, Q-Sepharose, and Mono S, and FPLC-gel filtration on Superdex 75. The protease functioned at an optimum pH of 7.0 and an optimum temperature of $50^{\circ}C$. It was also noted that the protease demonstrated a proteolytic activity of 1,756 U/mg toward casein. The $K_m$ of the purified protease toward casein was 6.36 mg/ml at a pH of 7.0 and with a temperature of $37^{\circ}C$, whereas the $V_{max}$ was 9.11 ${\mu}g\;ml^{-1}\;min^{-1}$. The activity of the protease was adversely affected by EDTA-2Na, suggesting that it is a metalloprotease. PMSF, EGTA, aprotinin, and leupeptin exerted no striking inhibitory effect. The activity of the protease was enhanced by $Fe^{2+}$, but was curtailed by $Cd^{2+}$, $Cu^{2+}$, $Hg^{2+}$, $Pb^{2+}$, $Zn^{2+}$, and $Fe^{2+}$ ions. The protease also exhibited inhibitory activity against HIV-1 reverse transcriptase with an $IC_{50}$ value of 4.00 ${\mu}M$. The $IC_{50}$ values toward hepatoma Hep G2 and leukemia L1210 cells in vitro were 4.99 ${\mu}M$ and 3.67 ${\mu}M$, respectively.

Purification, Partial Characterization, and Immunoassay of Vitellogenin from Marbled Sole (Limanda yokohamae)

  • Kim Dae-Jung;Jung Jee-Hyun;An Cheul-Min;Jee Young-Ju;Min Kwang-Sik;Kim Yoon;Han Chang-Hee
    • Fisheries and Aquatic Sciences
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    • v.8 no.4
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    • pp.213-219
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    • 2005
  • Vitellogenin (VTG) was purified from the blood plasma of estradiol-17$\beta$ ($E_2$)-treated male marbled sole (Limanda yokohamae) using gel filtration and anion exchange chromatography. The purity of the marbled sole VTG (msVTG) was confirmed by polyacrylamide gel electrophoresis (SDS-PAGE) and N-terminal amino acid sequencing. The purified msVTG was used to produce monoclonal and polyclonal antibodies in mice and rabbits, respectively, and the specificity of the polyclonal antisera for msVTG was confirmed by Western blot analysis. The antibodies cross­reacted with a protein of molecular mass approximately 160 kDa in the plasma samples of mature female marbled sole. No cross-reactivity was observed with the plasma of male fish. A direct non-competitive sandwich enzyme-linked immunosorbent assay (ELISA) was developed using the monoclonal anti-msVTG and polyclonal anti-msVTG antibodies, with purified msVTG as the standard protein. The values of the intra- and inter-assay variations were within the ranges of $8.l-9.8\%$ and $8.5-12.2\%$, respectively. The sensitivity was about 0.3 ng/mL. Serial dilutions of plasma from mature female sole reacted with the msVTG-antibodies in the sandwich ELISA, whereas the plasma from male fish did not. The results indicate that the maturation status of female marbled sole can be identified using a sandwich ELISA for msVTG.

Chemical and ecological studies of grasslands and soils of the Young-dong in Seoul, Korea (영동지구의 야초지와 토양에 대한 화학적, 생태학적 연구)

  • 박봉규;한진순
    • The Korean Journal of Ecology
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    • v.3 no.1_2
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    • pp.1-10
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    • 1980
  • This study was carried out to find out the correlation between plant groups and environomental gradient in the native grassfild near Kang Nam Express Terminal from August 26 to september 15, 1977. The correlations among plant species were calculated by using the method of x2 and the various relationships among soil environmental gradients were grouped by means of correlation coefficient. As a result, both the species and the soil environmental factors could be subdivided into four groups, and the four groups of the native grassfield were turned out to be correlated with the four groups of soil environmental factors. The first plant group (Panicum dichotomiflorum, Chenopodium album, Bidens forndosa, Humulus japonicus, Alopeculus amurensis, Potentilla fragarioides, Centipeda minima, Beckmannia erucaeformis) was formed in the releve 1∼10 reagions, organic matter by Turins's method, Total N, Exchange Ca and total of Exchange K, Na, Kg, Ca were abundent. The second plant group (Digitaria sanguinalis, Paltulaca oleracea, Echinochlea macrocrovi, Cyperus amuricus) was formed in the relve 11∼18 reagions, where Exchange Ca and total of Exchange K, Ng, Mg, Ca were little but Exchange A1 and sand were aburdent. In relve 19∼20 reagions, with much of Exchange Ca, total of Exchange K, Na, Mg, Ca, pH and a little Exchange Al, was found the third plant group (Pycreus sanguinoloentus, Cyperus amuricus, Ludwigia prostrata, Echinochloa echinata, Polygonum thunbergii, Salix glandulosa seedling) The fourth plant group (Cassia nomame, Polygnoum perfoliatum, Setaria viridis, Glocien ussuriensis, Lactuca indica, setaria glauca, Artemisia capill aris, Artmisia asiatica, Calamagrostis arundinacea, Kummerowia striata, Denothera odorata, Artemisia japonica, Carey laceolata) was formed in relve 21∼25 reagions, where organic matter by loss on ignition, Exchange Mg and Maximum water holding capacity were abundent but sand was a little. Thus it can be concluded that there is a close relationship between the vegetation group clasified by meaus of x2 and the environtal factors of soil devided by means of correlations coefficient.

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