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Trends in Rapid Detection Methods for Marine Organism-derived Toxins (해양 생물 유래 독소의 나노 기술 기반 신속 진단법 개발 동향)

  • Park, Chan Yeong;Kweon, So Yeon;Moon, Sunhee;Kim, Min Woo;Ha, Sang-Do;Park, Jong Pil;Park, Tae Jung
    • Journal of Food Hygiene and Safety
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    • v.35 no.4
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    • pp.291-303
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    • 2020
  • Marine organism-derived toxins have negative effects not only on human health but also in aquaculture, fisheries, and marine ecosystems. However, traditional analytical methods are insufficient in preventing this threat. In this paper, we reviewed new rapid methods of toxin detection, which have been improved by adopting diverse types of nanomaterials and technologies. Moreover, we herein describe the main strategies for toxin detection and their related sensing performance. Notably, to popularize and commercialize these newly developed technologies, simplifying the process of pre-treating real samples real samples is very important. As part of these efforts, numerous studies have reported pretreatment methods based on the antibody-immobilized magnetic nanoparticles, and some cases have applied nanoparticles to enhance the sensing performance by utilizing the intrinsic catalytic activity. Furthermore, some reports have introduced fluorescent nanoparticles, such as quantum dots, to represent the lower detection limits of conventional enzyme-based colorimetric methods and lateral flow assays. Some studies using electrochemical measurements based on aptamer-nanoparticle complexes have also been announced. In addition, as the response to new toxins generated by changes in the marine environment is still lacking, further research on diagnostic and detection is also greatly needed for these kinds of marine toxins and their derivatives.

Kinetic Studies of Lactic Acid Fermentation (Part 3) Effect of Phenol Derivatives on Fermentation (유산균발효에 관한 동력학적 연구 (제3보) 발효에 미치는 Phenol 유도체의 영향)

  • LEE Keun-Tai;YANG Hyeun-Suk
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.14 no.4
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    • pp.212-216
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    • 1981
  • The growth of Lactobacillus bulgaricus treated with vanillin, ortho-vanillin and guaiaco1 was studied on synthetic medium in mechanically agitated chemostat culture, The exponential-phase growth rate exhibited a maximum at the cells treated with 50 ppm vanillin. That stimulation, however, appears to be an effect on growth rate rather than total cell growth. And the others were inhibited by the chemicals. Much greater inhibition in growth of the cells treated with 100 ppm of each chemical than oars treated with 50 ppm was observed after 25 hour fomentation. For aerobic microbes, the alcohol dehydrogenase reaction is enhanced for the reproduction of NAD, which consequently cause to stimulate fermentation. For micro-aerophilic microbes , however, the same effect was not observed at the present study at least in the case of cell concentration. However except f or one treated with 50 ppm vanillin the same effect was observed in the case of growth is to. From the result using the glucose as a substrate, it was found that the cell concentrations measured in terms of ultimate optical density (UOB/ml), were 0.96 and 0.92, when treated with 50 and 100 ppm vanillin; 0.40 and 0.45 when treated with ortho-vanillin 50 and 100 ppm: 0.49 and 0.47, when treated with guaiacol 50 and 100 ppm. The specific growth rates were 0.44, 0.15, 0.25, 0.29, 0.37, and 0.34; the specific production rates wire 0.33, 0.15, 0.16, 0.22, 0.28, and 0.26 and the glucose concentrations (g/1) after 25 hour fermentation were 23.5, 32.8, 31.5, 29.5, 28.0 and 28.8, these all in the same sequences as the first.

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A Novel Chenodeoxycholic Derivative HS-1200 Enhances Radiation-induced Apoptosis in Human MCF-7 Breast Cancer Cells (담즙산 합성유도체(HS-1200)가 인체 유방암 세포주(MCF-7)에서 유도하는 방사선 감작 효과)

  • Lee Hyung Sik;Choi Young Min;Kwon Hyuk Chan;Song Yeon Suk
    • Radiation Oncology Journal
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    • v.22 no.2
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    • pp.145-154
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    • 2004
  • Purpose : To examine whether a synthetic bile acid derivatives (HS-1200) sensitizes the radiation-induced apoptosis in human breast cancer cells (MCF-7) and to investigate the underlying mechanism. Materials and Methods : Human breast cancer cells (MCF-7) in exponential growth phase were treated with HS-1200 for 24 hours at 37$^{\circ}C$ with 5$\%$ CO$_{2}$ in air atmosphere. After removal of HS-1200, cells were irradiated with 2$\~$8 Gy X-ray, and then cultured Ii drug-free media for 24-96 hours. The effect of radiation on the clonogenicity of MCF-7 cells was determined with clonogenic cell survival assay with 16$\mu$M of HS-1200. The induction of apoptosis was determined using agarose gel electrophoresis and Hoechst staining. The expression level of apoptosis-related molecules, such as PARP, Bax, Bcl-2, Bak and AIF, were assayed by Western blotting analysis with 40$\mu$M of HS-1200 combined with 8 Gy irradiation. To examine the cellular location of cytochrome c, bax and AIF immunofluorescent stainings were undertaken. Results : Treatment of MCF-7 cells with 40$\mu$M of HS-1200 combined with 8 Gy irradiation showed several changes associated with enhanced apoptosis by agarose gel electrophoresis and Hoechst staining. HS-1200 combined with 8 Gy irradiation treatment also enhanced production of PARP cleavage products and increased Bax/Bcl-2 ratio by Western blotting. Loss of mitochondrial membrane potential ($\Delta$$\psi$$_{m}$) and increased cytochrome c staining indicated that cytochrome c had been released from the mitochondria in HS-1200 treated cells. Conclusion : We demonstrated that combination treatment with a synthetic chenodeoxycholic acid derivative HS-1200 and irradiation enhanced radiation-induced apoptosis of human breast cancer cells (MCF-7). We suggest that the increased Bax/Bcl-2 ratio In HS-1200 co-treatment group underlies the increased radio sensitivity of MCF-7 cells. Further futures studies are remained elusive.

Regulatory Characterization of xylA Promoter Region in Escherichia coli (대장균의 xylA 프로모터 영역의 조절 특성)

  • Kang, Byung-Tae;Roh, Dong-Hyun;Joo, Gil-Jae;Rhee, In-Koo
    • Applied Biological Chemistry
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    • v.39 no.6
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    • pp.443-448
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    • 1996
  • In order to investigate the function of xylA promoter(Pxyl) as regulatory region Pxyl-lacZ fusion gene was constructed by the insertion of xylA promoter to the multiple cloning site of upstream of lacZ gene in a multicopy numbered plasmid pMC1403 containing promoterless lac operon, which was designated pMCX191, and Pxyl-lacZ fragment from pMCX191 was inserted to low copy numbered plasmid pLG339, designated pLGX191. The expressions of ${\beta}-galactosidase$ in these recombinant plasmids containing Pxyl-lacZ fusion gene were induced strongly by the addition of xylose, repressed by the addition of 0.2% glucose in the presence of xylose. The catabolite repressions were derepressed by the addition of 1 mM cAMP as same as native xylA gene. The fragment of xylA promoter was partially deleted from the upstream of xylA promoter by exonuclease III to investigate the regulation site of xylA promoter and the degrees of deletion derivatives of xylA promoter were analyzed by the DNA base sequencing. By the investigations of the induction by xylose, repression by glucose and derepression by cAMP on xylose isomerase production, the regulation site of xylA promoter may be located in segment between -165 and -59 bp upstream from the initiation site of xylA translation.

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Analysis on the Components in Stem of the Lespedeza bicolor (싸리(Lespedeza bicolor) 줄기의 성분 분석)

  • Lee, Yang-Suk;Joo, Eun-Yong;Kim, Nam-Woo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.8
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    • pp.1246-1250
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    • 2005
  • This study was carried out to analyze the components of the stem of bush clover, Lespedeza bicolor to obtain basic data on the studies of the physiological function and a possibility of utilizing L. bicolor. General components of L. bicolor were 82.35$\%$ carbohydrate, 13.38$\%$ moisture, 2.80$\%$ crude protein, 0.86$\%$ crude ash and 0.61$\%$ crude fat. The content of reducing sugar was 147.58 mg/100 g. The total amount of free sugar was 52.4 mg/100 g that was composed of 31.0 mg/100 g fructose, 16.7 mg/100 g glucose and 4.7 mg/100 g sucrose. In the results of mineral analysis, the content of Ca was the highest (472.97 mg/100 g) and followed by 206.70mg/100 g of K. The total amount of hydrolyzed amino acid was 148.95 mg/100 g that consist of 31.74 mg/100g of essential amino acid and 117.24 mg/100 g of non-essential amino acid. Total free amino acids were contained 106.39 mg/100 g that was composed of 8.41 mg/100 g essential amino acids and 97.98 mg/100 g the non-essential, and proline (62.92 mg/100 g) was the highest that account for 59.8$\%$ of total free amino acids. Total content of amino acid derivatives was 30.01 mg/100g and that of $\gamma$ -aminoisobutyric acid was the highest as 12.57 mg/100 g among them.

Ginsenoside derivatives and quality characteristics of fermented ginseng using lactic acid bacteria (유산균을 이용한 발효인삼의 ginsenoside 유도체 및 품질특성)

  • Kang, Bok-Hee;Lee, Kun-Jae;Hur, Sang-Sun;Lee, Dong-Sun;Lee, Sang-Han;Shin, Ki-Sun;Lee, Jin-Man
    • Food Science and Preservation
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    • v.20 no.4
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    • pp.573-582
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    • 2013
  • This study was done in order to investigate the bioconversion of ginsenoside, as well as the quality characteristics of fermented ginseng, by using lactic acid bacteria. Quality characteristics such as the thin layer chromatography(TLC) pattern, ginsenosides, total phenolic content, electron donating ability, and total sugar of fermenting ginseng and red ginseng were analyzed. The ginsenoside Rg2r, Rh2s and Rh2r of the fermented ginseng and red ginseng for 65 hours at a temperature of $37^{\circ}C$ were not detected. The ginsenoside Rg1 and Re contents have decreased, while the Rh1, Rg2s, Rd, Rg3r, and Rg3s have increased due to fermentation. The ginsenoside Rg3 of the fermented red ginseng has increased and the contents were $114.83{\sim}131.68{\mu}g/mL$ (control $104.56{\mu}g/mL$). The total phenolic content and electron donating ability of the red ginseng have totally decreased after 7 days of fermentation. The total phenolic contents of the fermented ginseng and red ginseng with different lactic acid bacteria did not show any tendency as different strains. The electron donating ability of the fermented ginseng has increased; however, the electron donating ability of the red ginseng has decreased. The total sugars of the fermented ginseng and red ginseng with different lactic acid bacteria have also decreased.

The Crystal Structure of Cholesteryl Carbonates (콜레스테롤 카보네이트 결정구조에 관한 연구)

  • 박영자;신정미
    • Korean Journal of Crystallography
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    • v.7 no.1
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    • pp.8-19
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    • 1996
  • Cholestryl Methyl and Propyl Carbonate(CH3OCOOC27H45, C3H7OCOOC27H45) are monoclinic, space group P21, with a=17.014(1), b=7.682(1), c=10.612(1)Å, β=103.05(1)°, Z=2, V=1351.16Å3, Dc=1.09 g/cm3 for methyl carbonate, and with a=13.683(1), b=11.864(2), c=18.904(2)Å, β=106.30(1)°, Z=4, V=2945.4Å3, Dc=1.06 g/cm3, Dm=1.06 g/cm3 for propyl carbonate. The intensity data were collected on an Enraf-Nonius CAD-4 diffractometer with a graphite monochromated Cu-Kα radiation. The structure was solved by direct methods and refined by full matrix least-squares methods. The final R factor was 0.051 for 2323 observed reflections for methyl carbonate and 0.074 for 3323 observed reflections for propyl carbonate. Compared with other cholesteryl derivatives, the cholesteryl ring and tail region of the molecules are normal. The molecules are stacked in clearly separated layers. At center of the layer, there are cholesteryl-C(17) side chain interactions. The interface region between layers is occupied by the loosely packed methyl carbonate chains. The structure of cholesteryl propyl carbonates have two propyl carbonates have two molecules(A, B) that are not related by crystal symmetry and have their tetracyclic system almost parallel to each other. Cholesteryl-cholesteryl interactions between symmetry related A-molecules, and cholesteryl-C(17) side chain interactions between symmetry related B-molecules occur at the center of the layers and these molecules stack along 2₁ screw axes. There are also C(17)chain-carbonate chain and C(17)chain-C(17)chain interactions in the interface region between layers. There is efficient packing between cholesteryl ring systems in propyl carbonates. Temperature ranges of cholesteric mesophases of cholesteryl alkyl cargonates are narrow for methyl, pentyl and hexyl carbonates, and rather broader for ethyl and propyl carbonates. Cholesteryl-isotropic transitions change very little with chain length.

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The effects of enamel matrix derivatives on the proliferation and gene expression of PDL fibroblast, $SaOs_2$ cells and Cementum derived cells (법랑기질유도체가 치주인대세포, 불멸화 조골세포, 백악질 유래세포의 증식과 유전자 발현에 미치는 영향)

  • Jeong, Yoo-Jee;Kim, Kyoung-Hwa;Kim, Tae-Il;Seol, Yang-Jo;Ku, Young;Lee, Hae-Jun;Rhyu, In-Chul;Chung, Chong-Pyoung;Han, Soo-Boo;Lee, Yong-Moo
    • Journal of Periodontal and Implant Science
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    • v.35 no.2
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    • pp.321-333
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    • 2005
  • 1. 목적 in vitro 상에서 법랑기질유도체가 치주인대섬유아세포, 불멸화 조골세포와 백악질 유래세포의 증식과 유전자 발현에 미치는 영향을 알아보고자 하였다. 2. 연구방법 및 재료 <세포증식 연구> 교정을 목적으로 발거한 치아에서 분리, 배양한 치주인대섬유아세포와 백악질유래세포, 그리고 $SaOs_2$ 세포를 이용하였다. 법랑기질유도체가 세포 증식에 미치는 영향을 알아보기 위해, 35 mm Petri dish에 dish 당 $5{\times}10^3$ 개의 세포를 접종하였다. 대조군은 1% 항생제와 10% FBS를 포함한 DMEM 배지를 이용했고, 5mM 초산을 첨가한 군과 첨가하지 않은 두 개의 대조군이 이용되었다. 실험군은 100 ${\mu}g/ml$의 법랑기질유도제를 첨가한 군과 100 ${\mu}g/ml$의 법랑기질유도체와 5 mM의 초산을 첨가한 2개의 실험군이 이용되었다. 각 군은 세 개의 배양접시에 행해졌고, 1, 3, 8일에 세포의 수를 각각 측정하였다. 결과는 repeated measures ANOVA로 통계 처리하였다. <유전자 발현 연구> 각 세포의 형질 특성을 알아보기 위해 RT-PCR을 실시하여 조골세포 분화 표식자와 연관된 Human collagen type I(COL I), human osteopontin(OP), human osteocalcin(OC), human alkaline phosphatase(ALP)와 human bone sialoprotein(BSP)의 mRNA 발현을 실험 1, 3, 8일에 걸쳐, 세 군의 차이를 비교 관찰하였다. 3. 결과 <세포증식 연구> 치주인대세포와 백악질유래세포, 그리고 $SaOs_2$ 세포의 증식은 법랑기질유도체에 의해 영향을 받지 않았다. 대조군과 초산이 포함된 대조군 그리고 법랑기질유도체와 초산이 포함된 실험군에서 유의할 만한 세포 수의 차이가 실험 기간 1, 3, 8일에 걸쳐 나타나지 않았다(p<0.05). <유전자 발현 연구> ALP와 COL I은 세 군의 세포에서 모두 발현되었고, 발현 정도는 EMD에 영향을 받지 않았다. OC은 세 군에서 모두 비교적 약하게 발현되었고, 특히 $SaOs_2$ cell과 백악질유래세포에서 약하게 발현되었다. EMD는 OC의 발현정도를 약하게 하였다. OP은 백악질유래세포에서 1, 3, 8일에 걸쳐 EMD 유무에 관련 없이 발현되지 않았다. 그러나 치주인대세포와 $SaOs_2$ cell에서는 강하게 발현되었다. BSP는 치주인대세포와 $SaOs_2$ cell에서 1, 3, 8일에 걸쳐 비교적 고르게 발현되었다. EMD 배지에서 배양된 백악질유래세포는 8일에는 BSP가 발현되지 않았다. 4 결론 이번 실험 결과에 의하면 법랑기질유도체는 치주인대세포, 불멸화 조골세포와 백악질 유래세포의 증식에 있어 유의성 있는 효과를 나타내지 않았다. 그러나, 유전자 발현에 있어서는, 치주인대세포와 백악질유래세포, 그리고 $SaOs_2$ 세포 모두에서 OC mRNA의 발현을 억제하는 효과를 나타내었다. EMD는 세포의 증식에는 영향을 미치지 않지만, 유전자 발현에 있어 일부 영향을 미치는 것으로 보인다. 법랑기질유도체가 세포의 증식과 유전자 발현에 미치는 영향은 배양된 세포의 형질특성, 배양환경, 배양일수 등에 따라 달라질 수 있다. 그러므로 법랑기질유도체가 in vitro 상에서 세포에 미치는 영향은 보다 정량화된 연구가 필요하다.

Characterization of an Extracellular Xylanase from Bacillus sp. HY-20, a Bacterium in the Gut of Apis mellifera (꿀벌(Apis mellifera)의 장내 세균인 Bacillus sp. HY-20이 분비하는 Xylanase의 특성)

  • Lee, Lan-Hee;Kim, Do-Young;Han, Mi-Kyoung;Oh, Hyun-Woo;Ham, Su-Jin;Park, Doo-Sang;Bae, Kyung-Sook;Sok, Dai-Eun;Shin, Dong-Ha;Son, Kwang-Hee;Park, Ho-Yong
    • Korean Journal of Microbiology
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    • v.45 no.4
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    • pp.332-338
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    • 2009
  • A xylan-decomposing bacterium, HY-20, was isolated from the gut of a honeybee, Apis mellifera, and identified as Bacillus sp. The extracellular GH11 xylanase (XylP) gene (687-bp) of strain HY-20 encoded a protein of 228 amino acids with a deduced molecular mass of 25,522 Da and a calculated pI of 9.33. The primary structure of XylP was 97% identical to that of B. pumilus xylanase (GenBank accession no.: AY526092) that has not been characterized yet. The recombinant His-tagged enzyme (rXylP) overexpressed in Escherichia coli BL21 harboring pET-28a(+)/xylP was purified to electrophoretic homogeneity by cation exchange and gel permeation chromatographies. The purified enzyme exhibited the highest catalytic activity toward birchwood xylan at pH 6.5 and $50^{\circ}C$ and retained approximately 50% of its original activity when pre-incubated at $55^{\circ}C$ for 15 min. The recombinant enzyme was completely inactivated by $Hg^{2+}$ (1 mM) and N-bromosuccinimide (5 mM), while its activity was slightly stimulated by approximately 10% in the presence of $Mn^{2+}$ (1 mM), $Fe^{2+}$ (1 mM), and sodium azide (5 mM). rXylP was able to efficiently degrade various polymeric xylose-based substrates but PNP-sugar derivatives and glucose-based polymers were not susceptible to the enzyme.

Enhanced Production of Oleanolic Acid by the Elicitation in Oldenlandia diffusa Suspension Cell Cultures (백화사설초의 현탁세포배양에서 Elicitation에 의한 Oleanolic acid 생산성 증대)

  • Lee Yong-Il;Kim Dong-Il
    • KSBB Journal
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    • v.19 no.6 s.89
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    • pp.471-477
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    • 2004
  • Oldenlandia diffusa is a Chinese medicinal herb with antitumor activity capable of suppressing the growth of some cancer cell lines. Oleanolic acid and ursolic acid are triterpenoid compounds that exist in Oldenlandia diffusa. Recently, these have been noted for anti-inflammatory, anti-cancer, and hepato-protective effects. Application of both plant growth regulators, 2,4-D and kinetin, was found to be essential for the initiation of callus and suspension cells. Leaf blades of Oldenlandia diffusa was transformed into callus on Schenk and Hildebrandt medium supplemented with 0.5 mg/L 2,4-D and 0.1 mg/L kinetin, while optimum initiation condition for suspension cells of Oldenlandia diffusa was determined to be 0.75 mg/L 2,4-D and 0.1 mg/L kinetin. Chromatographic separation of oleanolic acid from its derivatives was achieved using Rexchrom S5-100-ODS column. Analytical conditions for oleanolic acid were determined as follows: flow rate at 1.0 mL/min, UV length at 200 nm and mobile phase of $80\%$ acetonitrile and $20\%$ water. Production of secondary metabolites was found to be increased by the treatment with elicitors or signal transducers. The maximum production of oleanolic acid was 99.6 mg/L in cultures with 0.5 mM salicylic acid. It is 1.74 times higher than that of control.