• Title/Summary/Keyword: electro-phoresis

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Studies on the Insecticide Resistance of the German Cockroach(Blattella germanica L.). III. Comparison of Esterase Activity (바퀴(Blattella germanica L.)의 살충제 저항성에 관한 연구. 3. Esterase활성비교)

  • 방종렬;김정화;이형래
    • Korean journal of applied entomology
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    • v.32 no.3
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    • pp.265-270
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    • 1993
  • The German cockroach(Blattelia germanica) population~ were successIVely selected with ch\orpyrifos and permethrin during the six generations. The resulting resistant $R_{chtorpenfos}$(Rc) and $R_{permethnn}$(Rp) stra.ins were studied to investigate the esterase activity by spectrophotometer, filter parper test, and electrophoresis. Esterase-$\alpha$ activities by filter paper test showed 2.65 and LBZ times higher in the Rc and Rp strains than the susceptible strain, respectively. ln the spectrophoLometer method, the esterase activit18s to $\alpha$-and $\beta$-naphthyl acetate were increased 2.34 and 5.28 times in the Rc than susceptible strain, and 1.48 and 2.92 times in the Rp Limn susceptible stram, respectlvely. Zymogram patterns of eslerase isozyme by agarose gel electrophoresis showed totally five bands. The Rc and Rp strains showed two additive bands as, Est-2 and Est-3, which were not shown in the susceptible strain. but the Rp strain dId not show Est-5 bands which was COlumon in the Rc and susceptible strams.

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Role of Active Oxygens on DNA Damage by Low Molecular Carbonyl Compounds Derived from Maillard Reaction (Maillard 반응에서 유래되는 저분자 Carbonyl 화합물의 DNA손상작용에 대한 활성산소종의 역할)

  • 김선봉;박성준;강진훈;변한석;박영호
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.19 no.6
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    • pp.565-570
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    • 1990
  • The role of the active oxygens on plasmid DNA damage by carbonyl compounds derived from Maillard reaction was investigated. Plasmid DNA extracted from E. coli Hb1O1 was reacted with carbonyl compounds, such as glyoxal, methyl glyoxal, dihydroxyacetone, diacetyl, glyceraldehyde, glycolaldehyde and furfural with and without the active oxygen scavengers at 37$^{\circ}C$ for 6 hours, and then the degree of damage was determined by using 1 % agarose gel electro-phoresis. All of the carbonyl compounds except furfural caused to damage of DNA. Among these, glyoxal, methyl glyoxal and dihydroxyacetone markedly induced the damage of DNA. On the other hand, the DNA damage by the carbonyl compounds was greatly inhibited by catalase, superoxide dismutase and $\alpha$-tocopherol it is considered that the damage of DNA is due to active oxygens, such as singlet oxygen, hydrogen peroxide and superoxide anion generated during the autoxidation of carbonyl compounds.

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Physicochemical Properties and Dietary Effect of Glycoprotein from Sea Cucumber(Stichopus japonicus) (해삼 당단백질의 물리화학적 특성과 식이효과)

  • 류홍수;문정혜;유병진;문수경
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.25 no.2
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    • pp.240-248
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    • 1996
  • To clarify the potentiality of sea cucumbers as dietary food, the effects of those glycoprotein on dietary proteins and physicochemical properties of those proteins were studied. Crude glycoprotein was efficiently extracted using 20mM sodium phosphate beffer(pH 7.0) and by salting out with 80% ammoniym sulfate saturation. The fractions obtained through the DEAE-cellulose ion exchange chromatography was identified as glycoprotein by Schiff's reagent and SDS polyacrylanide gel electro-phoresis. The yields of each glycoprotein from the three kinds of sea cucumbers were 0.814(red), 0.184(blue) and 0.232(black) and the molecular weights of the glycoproteins subunits were ranged from 20,000 dalton(blue and black) to 29,000 dalton(red), respectively. The electrophoretic patterns of the glycoprotein isolates were similar to each other and any significant difference in amino acid pattern was observed. Predominant arnino scids were Asx(aspartic acid and asparagine) and Glx(glutamic acid and glutamine) ; in contrast, histidine and methionine were below 2% as compared to total amino acids. water holding capacities of the glycoprotein isolates from red, blue and black cucumbers were equally 100% and emulsion activities ranged from 53% to 64%. In addition the emulsion stabilities were 7.04, 1.37 and 2.44, respectively. In vitro digestibility of some proteins(casein, SPI and squid) was decreased as increasing the level of the freeze dried sea cucumber powder and glycoprotein isolates. But squid protein was not affected.

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Identification of a Protein Kinase using a FITC-labelled Synthetic Peptide in Streptomyces griseus IFO 13350 (형광 Peptide를 이용한 Streptomyces griseus IFO 13350의 인산화 단백질 동정)

  • 허진행;정용훈;김종희;신수경;현창구;홍순광
    • Microbiology and Biotechnology Letters
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    • v.30 no.3
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    • pp.235-240
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    • 2002
  • Streptomycetes is a group of Gram-positive soil bacteria that growas a branching vegetative mycelium leading to the formation of spores, and display a physiological differenti-ation related to the synthesis of many secondary metabolites including antibiotics. Their complex life cycle and multicellular differentiation require various levels of regulation and types of signal transduction systems including eukaryotic-type serine/threonine protein kinases and prokaryotic-type histidine/aspartic acid protein kinases. Akt kinase that was found in cells is a sorine/threonine kinase controlling signal pathway for multi-tude of important cellular events. The activation or inactivation of Akt kinase in the cell is one of the critical regulatory points to deliver cell proliferation, differentiation, survival or apoptosis signal. To find the regula-tory protein homologous to Akt in Streptomyces, the fluorescien-labeled synthetic peptide (FITC-TRRSR-TESIT) was designed from the consensus sequence of target proteins for Akt kinase. From the difference of the mobility between the nonphosphorylated and phosphorylated synthetic peptides on Agarose gel electro-phoresis, the Akt-phosphorylating activity was monitored. The cell-free extract prepared from Streptomyces griseus IFO 13350 and the Akt homologous protein was purified by ammonium sulfate fractionation and many steps of column chromatographies such as, DEAE-Sepharose, Mono Q, Resource Phenyl-Soporose and Gel permeation column chromatographies. As a result, the protein phosphorylating the fluorescien-labeled Akt substrate was identified and it's molecular weight was estimated as 39 kDa on SDS-PAGE.

Interleukin-8-like chemotactic factor from feline peripheral blood mononuclear cells cultured with egg white derivatives (계난백유래물질로 배양한 고양이 말초혈액 단핵구세포에서 분비되는 interleukin- 8 양(樣) 유주성인자)

  • Lee, Jae-kwon;Yang, Mhan-pyo
    • Korean Journal of Veterinary Research
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    • v.40 no.2
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    • pp.393-401
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    • 2000
  • The feline chemotactic factor(s) for polymorphonuclear cells (PMN) in culture supernatant from mononuclear cells (MNC) treated with egg white derivatives (EWD) were examined. Culture supernatant from MNC treated with EWD and human recombinant (hr) IL-8 remarkably enhanced chemo-taxis of feline PMN. To investigate feline chemotactic factor(s), gel electrophoresis was performed with culture supernatant from MNC treated with EWD under denaturing (18% loading gel/5% stacking gel) and nondenaturing (12.5% loading gel/5% stacking gel) condition. Hr IL-8 and culture supernatant from MNC treated with EWD yielded a distinct band in a molecular weight, 6 to 8 kDa. Eluted solution from gel slices of 6 to 8 kDa band in denaturing condition also enhanced feline PMN chemotaxis. These chemotactic activities of feline PMN induced by culture supernatant from MNC treated with EWD, hr IL-8 and eluted solution were inhibited in a dose-dependent manner by rabbit anti-feline polyclonal IgG (RAF pIgG) and monoclonal antibody (mAb) against hr IL-8. RAF pIgG also showed a binding activity with hr IL-8, suggesting that RAF pIgG against feline IL-8-like chemotactic factor(s) had cross-reactivity with human IL-8. These results suggested that feline MNC treated with EWD might release feline IL-8-like chemotactic factor(s) with a molecular weight, 6 to 8 kDa, which induces the chemotaxis of feline PMN.

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Radiation-induced Apoptosis is Differentially Modulated by PTK Inhibitors in K562 Cells (K562 백혈병 세포주에서 방사선에 의해 유도되는 Apoptosis에 미치는 PTK Inhibitors의 영향)

  • Lee Hyung Sik;Moon Chang Woo;Hur Won Joo;Jeong Su Jin;Jeong Min Ho;Lee Jeong Hyeon;Lim Young kin;Park Heon Joo
    • Radiation Oncology Journal
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    • v.18 no.1
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    • pp.51-58
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    • 2000
  • Purpose :The effect of PTK inhibitors (herbimycin A and genistein) on the induction of radiation-induced apoptosis in Ph-positive KS62 leukemia cell line was investigated. Materials and Methods :K562 cells in exponential growth phase were irradiated with a linear accelerator at room temperature. For 6 MV X-ray irradiation and drug treatment, cultures were initiated at 2×106 cells/mL. The cells were irradiated with 10 Gy. Stock solutions of herbimycin A and genistein were prepared in dimethyl sulphoxide (DMSO). After incubation at 37$^{\circ}C$ for 0$\~$48 h, the extent of apoptosis was determined using agarose gel electrophoresis and TUNEL assay. The progression of cells through the cell cycle after irradiation and drug treatment was also determined with flow cytometry. Western blot analysis was used to monitor bel-2, bel-X$_{L}$ and bax protein levels. Results :Treatment with 10 Gy X-irradiation did not result in the induction of apoptosis. The HMA alone (500 nM) also failed to induce apoptosis. By contrast, incubation of K562 cells with HMA after irradiation resulted in a substantial induction of nuclear condensation and fragmentation by agarose gel electro-phoresis and TUNEL assay. Genistein failed to enhance the ability of X-irradiation to induce DNA fragmentation. Enhancement of apoptosis by HMA was not attributable to downregulation of the bel-2 or bel-X$_{L}$ anti-apoptotic proteins. When the cells were irradiated and maintained with HMA, the percentage of cells in G2/M phase decreased to 30$\~$40$\%$ at 48 h. On the other hand, cells exposed to 10 Gy X-irradiation alone or maintained with genistein did not show marked cell cycle redistribution. Conclusion : We have shown that nanomolar concentrations of the PTK inhibitor HMA synergize with X-irradiation in inducing the apoptosis in Ph (+) K562 leukemia cell line. While, genistein, a PTK inhibitor which is not selective for p210$^{bcr/abl}$ failed to enhance the radiation induced apoptosis in KS62 cells. It is unlikely that the ability of HMA to enhance apoptosis in K562 cells is attributable to bel-2 family. It is plausible that the relationship between cell cycle delays and cell death is essential for drug development based on molecular targeting designed to modify radiation-induced apoptosis.

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Comparison of Grain and Milling Properties between Waxy and Non-waxy Wheat (찰성밀과 보통밀간의 종실 및 제분특성비교)

  • Lee Choon-Ki;Nam Jung-Hyun;Kang Moon-Seok;Ku Bon-Chol;Park Kwang-Keun;Kim Jae-Cheol;Son Young-Koo;Park Jeong-Hwa;Lee Yeong-Ho;Son Jong-Rok;Min Young-Kyoo
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.50 no.6
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    • pp.428-434
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    • 2005
  • For the purpose to verify the grain and evaluate milling properties of Korean waxy wheat, com­parison analysis between waxy wheat lines and their respective maternal parents were performed. The waxy lines showed various grain yields of 4.76${\~}$5.79 t/ha depending on parentages, which were corresponding to $80{\~}96\%$ levels of their respective maternal parents. One thousand grain weights of waxy lines were also lighter than its respective part in its parentage by exhibiting 32.8${\~}$34.6 g compared to 32.9${\~}$45.2 g of their parents. Test weights of waxy lines and their parents were 720${\~}$798 g/l and 786${\~}$797 g/l, respectively. The proportions of the grains above 2.5 mm in width were higher in order of Keumgang, Olgeuru, Geuru, SW97134, Suwon 292, Woori, SW97105, and SW97110. Waxy lines exhibited low milling properties by showing the straight flour yields ranging from $61.8\%$ to $67.1\%$ compared with the yields of their parents ranging from $66.1\%$ to $72.5\%$; the waxy lines were significantly lower in first break flour (Bl) and first reduction flour (Rl) yields in the Buller test mill, while significantly higher in the yields of second and third reduction flour (R2 and R3) than the respective ones of their parent wheat.

A Study of the Distribution of Listeria spp. in Fresh Agricultural Products Distributed in the Busan Area, the Republic of Korea (부산지역에서 유통되는 신선농산물 중 리스테리아균 분포에 관한 연구)

  • Youn-ju Ok;Young-hee Kwon;Hye-sun Hwang;Ye-jee Byun;Ji-young Park;Byung-jun Kim
    • Journal of Food Hygiene and Safety
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    • v.39 no.1
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    • pp.9-15
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    • 2024
  • This study was performed to survey the distribution of Listeria spp. in fresh agricultural products in the Busan area, the Republic of Korea, from January to November 2022. We investigated the pathogenicity and epidemiological relationships by tracing isolated strains using polymerase chain reaction and pulsed-field gel electro-phoresis (PFGE) methods. Forty cases of Listeria spp. were detected in the 210 samples of fresh agricultural products analyzed. Four species, Listeria rocourtiae, L. innocua, L. grayi, and L. monocytogenes were detected only in green vegetables (lettuce, perilla leaps) and the others (L. innocua, L. monocytogenes, and L. grayi) were detected in enoki and oyster mushrooms. L. innocua was detected in 22 samples and L. grayi in six samples. L. monocytogenes, which causes foodborne diseases, was only detected in enoki mushrooms and the strains that were isolated had genes responsible for the pathogenicity of listeriosis (iap, prfA, inlA, inlC, inlJ, and hly). To investigate the genetic similarity and contamination route of L. monocytogenes, serotyping and PFGE were conducted for 12 strains isolated from fresh agricultural (10 strains) and poultry (2 strains) products distributed at a market in the Busan area. Two serotypes (1/2a, 1/2b) were detected in strains isolated from the agricultural and poultry products, but serotype 1/2b was only detected in strains isolated from agricultural products. PFGE analysis showed index of similarity values of 45.7 to 100% and the same patterns were represented in isolates from some enoki mushrooms. These isolates had the same serotypes and showed significant epidemiological relationships.