• Title/Summary/Keyword: Isothiocyanate

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Apolipophorin-III uptake by the last larval fat body in the wax moth Galleria mellonella (꿀벌부채명나방 종령 유충 지방체에 의한 아포리포포린-III의 흡수)

  • Yun, Hwa-Kyung
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.14 no.8
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    • pp.4106-4110
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    • 2013
  • Apolipophorin-III (apoLp-III) was isolated and purified from the last larval hemolymph of Galleria mellonella by the KBr gradient ultracentrifugation and gel chromatography (Sephadex G-100). In this paper, we examined that apoLp-III is taken up into the last larval fat bodies in Galleria mellonella. The last larval fat body tissues were incubated at room temperature for 30 min with fluorescein isothiocyanate (FITC)-labeled apoLp-III (FITC-apoLp-III). Fluorescein microscopy and sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) revealed that the last larval fat body tissues internalize FITC-apoLp-III. The results show that the apoLp-III is taken up by the last larval fat body.

Assessment of Lipopolysaccharide-binding Activity of Bifidobacterium and Its Relationship with Cell Surface Hydrophobicity, Autoaggregation, and Inhibition of Interleukin-8 Production

  • Park, Myeong-Soo;Kim, Min-Jeong;Ji, Geun-Eog
    • Journal of Microbiology and Biotechnology
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    • v.17 no.7
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    • pp.1120-1126
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    • 2007
  • This study was performed to screen probiotic bifidobacteria for their ability to bind and neutralize lipopolysaccharides (LPS) from Escherichia coli and to verify the relationship between LPS-binding ability, cell surface hydrophobicity (CSH), and inhibition of LPS-induced interleukin-8 (IL-8) secretion by HT-29 cells of the various bifidobacterial strains. Ninety bifidobacteria isolates from human feces were assessed for their ability to bind fluorescein isothiocyanate (FITC)-labeled LPS from E. coli. Isolates showing 30-60% binding were designated LPS-high binding (LPS-H) and those with less than 15% binding were designated LPS-low binding (LPS-L). The CSH, autoaggregation (AA), and inhibition of LPS-induced IL-8 release from HT-29 cells of the LPS-H and LPS-L groups were evaluated. Five bifidobacteria strains showed high levels of LPS binding, CSH, AA, and inhibition of IL-8 release. However, statistically significant correlations between LPS binding, CSH, AA, and reduction of IL-8 release were not found. Although we could isolate bifidobacteria with high LPS-binding ability, CSH, AA, and inhibition of IL-8 release, each characteristic should be considered as strain dependent. Bifidobacteria with high LPS binding and inhibition of IL-8 release may be good agents for preventing inflammation by neutralizing Gram-negative endotoxins and improving intestinal health.

Synthesis and Reactivity of Dinuclear Ni(II) Azido Complexes Containing Bithienylene or Terthienylene Bridging Ligands

  • Kim, Yong-Joo;Lee, Hyuck-Hee;Zheng, Zhen Nu;Lee, Soon-W.
    • Bulletin of the Korean Chemical Society
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    • v.32 no.9
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    • pp.3239-3244
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    • 2011
  • Dinuclear Ni(II)-thiophene halides, which contain linear bridging thienylenes, trans,trans-[$(PR_3)_2$(X)Ni-Y-Ni(X)$(PR_3)_2$] {X = Cl, Br; $H_2Y$ = 5,5'-dichloro-2,2'-bithiophene ($H_2bth$); $H_2tth$ = 5,5"-dichloro-2,2':5',2''-terthiophene ($H_2tth$)} were prepared by the oxidative addition of dihalobithiophene ($H_2bth$) or dihaloterthiophene ($H_2tth$) to [$Ni(COD)_2$] in the presence of tertiary phosphines. Subsequent reactions of $NaN_3$ with the dinuclear Ni(II)-thiophene chlorides gave the corresponding Ni(II)-azido complexes, trans,trans-[$(PR_3)_2(N_3)$Ni-Y-Ni$(N_3)(PR_3)_2$], whose reactivity toward trimethylsilyl pseudohalides such as trimethylsilyl isothiocyanates and cyanides was investigated. In addition, the reaction of trans-[$BrNi(PEt_3)_2-C_4H_2S-C_4H_2S$-CHO], a thienyl Ni(II) complex containing a terminal aldehyde group, with phosphonium ylide was examined.

Construction and Patterning of the biospecific affinity surfaces on gold using dendrimer

  • Hong, Mi-Yeong;Yun, Hyeon-Cheol;Kim, Hak-Seong
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.703-706
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    • 2000
  • We constructed a biospecific affinity surface using hyper-branched dendrimers on gold for biospecific recognition, and characterized the resulting surfaces by using confocal fluorescence microscopy. The dendrimer monolayer was firstly constructed on the mercaptoundecanoic acid SAM/Au with pentafluorophenyl ester activation and further functionalized with sulfo-NHS-biotin, an activated ester of biotin. To confirm the formation of biospecific affinity surface, FITC(fluorescein isothiocyanate)-labeled avidin was loaded onto the biotinylated dendrimer monolayer, and fluorescence images of the bound avidins were investigated with a confocal microscope. The constructed biospecific affinity surface showed a much more dense and uniform fluorescence compared to those from poly-L-lysine- and cystamine SAM-based affinity surfaces. For the dependency on the concentration of added FITC-labeled avidin on the affinity surface, derived fluorescence could be detectable from as low as $1{\mu}g/ml$, and intensified up to $50{\mu}g/ml$. Further reaction of FITC-labeled avidin layer with TMR(tetramethylrhodamine)-biocytins resulted in the efficient FRET(fluorescence resonance energy transfer) phenomenon. As an extension of the study, we attempted a patterning of the affinity surfaces on gold by microcontact printing. Fluorescence of the patterned surface demonstrated that FITC-labeled avidin molecules were specifically bound to the biotinylated patches.

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Microwave Assisted Synthesis of 1,3,4-Oxadiazole/Thiohydantoin Hybrid Derivatives via Dehydrative Cycliztion of Semicarbazide

  • Yang, Seung-Ju;Lee, Jae-Min;Lee, Gee-Hyung;Kim, NaYeon;Kim, Yong-Sang;Gong, Young-Dae
    • Bulletin of the Korean Chemical Society
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    • v.35 no.12
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    • pp.3609-3617
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    • 2014
  • A series of compounds containing both 1,3,4-oxadiazole and thiohydantoin were synthesized as a promising scaffold for application in medicinal chemistry. The key step of the synthesis is a microwave-assisted cyclization of semicarbazides possessing a thiohydantoin moiety at one of the acyl termini using $POCl_3$ as a dehydrating reagent. A wide range of semicarbazides were prepared through the substitution of hydrazides with an N-acylated thiohydantoin derived from the cyclization of the corresponding isothiocyanate with an amino acid and subsequent N-acylation of the resultant thiohydrantion. Consequently, the 58 number of 1,3,4-oxadiazole derivatives having a thiohydantoin substituent were prepared in good overall yields.

Suppressive Effects of Young Radish Cultivated with Sulfur on Growth and Metastasis of B16-F10 Melanoma Cells

  • Kim Soo-Jin;Kim Byung-Sam;Kyung Tae-Wook;Lee Sang-Chul;Rho Chi-Woong;Choi Kyung-Rak;Hwang Hae-Jun;Choi Hye-Seon
    • Archives of Pharmacal Research
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    • v.29 no.3
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    • pp.235-240
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    • 2006
  • The oral administration of extracts of young radishes cultivated with sulfur after intravenous tumor cell injection achieved a marked reduction of pulmonary colonization in mice. Treatment of the mice with extracts of young radish cultivated with sulfur did not show any increase in the number of CD8+ or NK T cells in the spleen, indicating no influence on host immunity. Sulforaphane, which could be a candidate for an active compound from young radishes cultivated with sulfur, inhibited cell growth of B16-F10 melanoma cells. In addition, extracts of the young radish cultivated with sulfur-fed group showed enhanced quinine reductase (QR) activities in the liver and lung and a slight increase of glutathione S-transferase (GST) activity in the liver. These results suggested that the administration of extracts of young radishes cultivated with sulfur suppressed pulmonary tumorigenesis, possibly due to increased activity of detoxification enzymes in the liver and lung, and partly due to cell cytotoxicity.

Correlation between Protein Methylation and Hepatotoxicity (단백질메칠화 반응과 간독성간의 상관관계)

  • 김재현;박창원;이주한;백윤기;문화회;홍성렬;이향우
    • Biomolecules & Therapeutics
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    • v.2 no.1
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    • pp.47-53
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    • 1994
  • The methylation response as well as the level of methyl donor substance, 5-adenosyl-L-methionine (SAM) has been suggested to be related to hepatotoxicity including hepatocarcinogenesis. But direct correlation between protein methylation and hepatotoxicity has not been established to the present. To observe relationship between protein methylation and short-term hepatotoxicity induced by chemical substances, the activities of protein methylase I and II (PM I, PM II) were examined in cytosolic fraction of SD rat treated orally with acetaminophen(AA), $\alpha$-naphtyl-isothiocyanate (ANIT) and tetracycline (TC) that was known to produce necrosis, cholestasis and steatosis respectively. To evaluate the degree of hepatotoxicity induced by each chemicals, we observed the serum levels of indicative parameters and histopathological alteration. In AA treated group, the activities of PM I were increased at 6, 12 hours after administration, prior to the appearance of the hepatotoxicity by clinical parameters. It was suggested that the levels of PM I were related with the initial stage of hepatotoxic mechanism induced by AA. In ANIT treated group, though most of clinical parameters were significantly increased at 24, 48 hours after administration, the activity of PM I was not changed, indicating that ANIT induced hepatotoxicity was not coupled to protein methylation.

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Mechanisms of Anticancer Activity of Sulforaphane from Brassica oleracea in HEp-2 Human Epithelial Carcinoma Cell Line

  • Devi, J. Renuka;Thangam, E. Berla
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.5
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    • pp.2095-2100
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    • 2012
  • Sulforaphane (SFN) an isothiocyanate formed by hydrolysis of glucosinolates found in Brassica oleraceae is reported to possess anticancer and antioxidant activities. In this study, we isolated SFN from red cabbage (Brassica oleraceae var rubra) and evaluated the comparative antiproliferative activity of various fractions (standard SFN, extract and purified SFN) by MTT assay in human epithelial carcinoma HEp -2 and and Vero cells. Probable apoptotic mechanisms mediated through p53, bax and bcl-2 were also examined. The SFN fraction was collected by HPLC, enriched for its SFN content and confirmed. Expression of apoptosis-related proteins was detected by western blotting and RT PCR. Results showed that Std SFN and purified SFN concentration found to have closer $IC_{50}$ which is equal to 58.96 microgram/ml (HEp-2 cells), 61.2 microgram/ml (Vero cells) and less than the extract which is found to be 113 microgram/ml (HEp-2 cells) and 125 microgram/ml (Vero cells). Further studies on apoptotic mechanisms showed that purified SFN down-regulated the expression of bcl-2 (antiapoptotic), while up-regulating p53 and Bax (proapoptotic) proteins, as well as caspase-3. This study indicates that purified SFN possesses antiproliferative effects the same as Std SFN and its apoptotic mechanism in HEp-2 cells could be mediated through p53 induction, bax and bcl-2 signaling pathways.

Rapid and Direct Detection of Vibrio vulnificus in Small Octopus (Octopus variabilis) Using Polymerase Chain Reaction

  • Choi, Sang-Ho;Lee, Jee-Yeon
    • Journal of Microbiology and Biotechnology
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    • v.5 no.4
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    • pp.181-187
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    • 1995
  • The cells of Vibrio vulnificus can be induced to the viable but nonculturable (VBNC) state by natural environmental parameters. The V. vulnificus cells in the VBNC state can not be recovered by ordinary laboratory techniques. This nonculturability could often hamper development of effective processing strategies to minimize the number of V. vulnificus in seafoods. Even with V. vulnificus cells in a culturable state, the length of time required to identify the bacteria in contaminated food by phenotyphic characterization may prevent appropriate in-time responses by public health agencies to infections of the bacteria. In the present study, we used polymerase chain reaction (PCR) to develop a rapid and direct detection method for V. vulnificus in small octopus (Octopus variabilis) which is consumed as a raw food in Korea. The region targeted was a 704-base pair (bp) portion of the hemolysin gene, vvhA, of V. vulnificus. The primers designed for PCR amplification were specific for all V. vulnificus sp. tested. Several methods were examined to extract total DNA directly from V. vulnificus seeded into the octopus homogenate and the guanidine isothiocyanate (CITC) method appeared to be most effective. From the octopus homogenate seeded by V. vulnificus at an initial level of $10^2$ CFU/ml of the homogenate and then incubated for 12 h, the targeted sequence was successfully amplified by PCR and the 704-bp DNA fragment was observed by gel electrophoresis. The total completion of this assay requires less than one day.

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Surface Modification of Magnetites Using Maltotrionic Acid and Folic Acid for Molecular Imaging

  • Selim, K.M.Kamruzzaman;Lee, Joo-Hee;Kim, Sun-Jung;Xing, Zhicai;Kang, Inn-Kyu;Chang, Yong-Min;Guo, Haiqing
    • Macromolecular Research
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    • v.14 no.6
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    • pp.646-653
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    • 2006
  • Highly hydrophilic, uniform, superparamagnetic and nontoxic maltotrionic acid (MA)-coated magnetite nano-particles (MAM) were prepared and characterized by TEM, DLS, XRD and VSM. MA was used to improve the biocompatibility, monodispersity and non-specific intracellular uptake of nanoparticles. Folic acid (FA) was subsequently conjugated to the MAM to preferentially target KB cells (cancer cells) that have folate receptors expressed on their surfaces and to facilitate nanoparticles in their transit across the cell membrane. Finally, fluorescence isothiocyanate (FITC) was added to the nanoparticles to visualize the nanoparticle internalization into KB cells. After the cells were cultured in a media containing the MAM and MAM-folate conjugate (FAMAM), the results of fluorescence and confocal microscopy showed that both types of nanoparticles were internalized into the cells. Nevertheless, the amount of FAMAM uptake was higher than that of MAM. This result indicated that nanoparticles modified with MA and FA could be used to facilitate the nanoparticle uptake to specific KB cells (cancer cells) for molecular imaging.