• Title/Summary/Keyword: Succinate

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Detection of Fungicidal Activities against Alternaria dauci Causing Alternaria Leaf Spot in Carrot and Monitoring for the Fungicide Resistance (당근검은잎마름병균 Alternaria dauci에 대한 살균제 효과 검정 및 병원균 집단에 대한 저항성 검정)

  • Do, Jiwon;Min, Jiyoung;Kim, Yongsu;Park, Yong;Kim, Heung Tae
    • Research in Plant Disease
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    • v.26 no.2
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    • pp.61-71
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    • 2020
  • With 32 fungicides, it was examined the inhibitory effects on the mycelial growth of Alternaria dauci KACC42997 causing Alternaria leaf blight of carrot. Showing the results of the agar dilution method, the fungicides belonging to C2, C5, G1, E2, and E3 group were excellent in inhibiting mycelial growth. Protective fungicides belonging to M group, except for iminoctadine tris-albesilate, and pyraclostrobin belonging to C3 group were effective in inhibiting spore germination of pathogens. The fungicides included into C2 group inhibiting succinate dehydrogenase activity and the G1 group inhibiting demethylase activity showed the excellent inhibitory effect on mycelial growth but the inhibitory effect of spore germination was very low. However, fluazinam belonging to C5 group was excellent in inhibiting spore germination as well as mycelial growth. Especially, when 100 ㎍/ml of fluxapyroxad belonging to the C2 group was treated, 47.1% of spore formation was inhibited on the medium. In comparison of the resistance factors of 3 fungicide groups, as G, C, and E group, in populations of A. dauci isolates collected from Gumi, Pyeongchang, and Jeju, resistance factor in the population of Jeju was the lowest. However, two isolates resistant to fludioxonil belonging to E2 group were found in the isolate group of Pyeongchang, and both showed cross-resistance to iprodione and procymidone.

TATA box binding protein and ribosomal protein 4 are suitable reference genes for normalization during quantitative polymerase chain reaction study in bovine mesenchymal stem cells

  • Jang, Si-Jung;Jeon, Ryoung-Hoon;Kim, Hwan-Deuk;Hwang, Jong-Chan;Lee, Hyeon-Jeong;Bae, Seul-Gi;Lee, Sung-Lim;Rho, Gyu-Jin;Kim, Seung-Joon;Lee, Won-Jae
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.12
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    • pp.2021-2030
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    • 2020
  • Objective: Quantitative polymerase chain reaction (qPCR) has been extensively used in the field of mesenchymal stem cell (MSC) research to elucidate their characteristics and clinical potential by normalization of target genes against reference genes (RGs), which are believed to be stably expressed irrespective of various experimental conditions. However, the expression of RGs is also variable depending on the experimental conditions, which may lead to false or contradictory conclusions upon normalization. Due to the current lack of information for a clear list of stable RGs in bovine MSCs, we conducted this study to identify suitable RGs in bovine MSCs. Methods: The cycle threshold values of ten traditionally used RGs (18S ribosomal RNA [18S], beta-2-microglobulin [B2M], H2A histone family, member Z [H2A], peptidylprolyl isomerase A [PPIA], ribosomal protein 4 [RPL4], succinate dehydrogenase complex, subunit A [SDHA], beta actin [ACTB], glyceraldehyde-3-phosphate dehydrogenase [GAPDH], TATA box binding protein [TBP], and hypoxanthine phosphoribosyltrasnfrase1 [HPRT1]) in bovine bone marrow-derived MSCs (bBMMSCs) were validated for their stabilities using three types of RG evaluation algorithms (geNorm, Normfinder, and Bestkeeper). The effect of validated RGs was then verified by normalization of lineage-specific genes (fatty acid binding protein 4 [FABP4] and osteonectin [ON]) expressions during differentiations of bBMMSCs or POU class 5 homeobox 1 (OCT4) expression between bBMMSCs and dermal skins. Results: Based on the results obtained for the three most stable RGs from geNorm (TBP, RPL4, and H2A), Normfinder (TBP, RPL4, and SDHA), and Bestkeeper (TBP, RPL4, and SDHA), it was comprehensively determined that TBP and RPL4 were the most stable RGs in bBMMSCs. However, traditional RGs were suggested to be the least stable (18S) or moderately stable (GAPDH and ACTB) in bBMMSCs. Normalization of FABP4 or ON against TBP, RPL4, and 18S presented significant differences during differentiation of bBMMSCs. However, although significantly low expression of OCT4 was detected in dermal skins compared to that in bBMMSCs when TBP and RPL4 were used in normalization, normalization against 18S exhibited no significance. Conclusion: This study proposes that TBP and RPL4 were suitable as stable RGs for qPCR study in bovine MSCs.

THE CYTOTOXIC EFFECTS OF GLASS-IONOMER CEMENT LINERS ON FIBROBLASTS IN HUMAN PULP (Glass-ionomer Cement 이장재의 세포독성에 관한 연구)

  • Na, Young-Min;Min, Byung-Soon;Choi, Ho-Young;Park, Sang-Jin;Choi, Gi-Woon
    • Restorative Dentistry and Endodontics
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    • v.18 no.2
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    • pp.261-276
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    • 1993
  • The purpose of this study was to evaluate for the cytotoxicity of glass-ionomer cement liners(GC liningcement, Ketac-bond, Vitrebond and Fuji lining LC) on the fibroblasts cultured from human pulp. The fibroblasts were cultured in DMEM-10% FBS medium. The measurement of pH, succinate dehydrogenase (SDH) activity test and $^{51}Chromium$ release test were performed. Viable cell count and $^{14}C$-leucine incorporation rate were evaluated following culture time of 2, 4 and 6 days. The results of this study were as follows : 1. The pH in all cements was to be neutralized as time elapsed, and Fuji lining LC was the lowest pH value among them. 2. SDH activity was more inhibited in GC lining cement and Vitrebond than Ketac-bond and Fuji lining LC with the setting process, and GC lining cement and Ketac-bond were reduced after 5 minute's setting and then elevated as time elapsed. 3. In SDH activity test following exposure time, the activity in Vitrebond, GC lining cement and Fuji lining LC was inhibited with increased exposure time, but it was fairly constant in Ketac-bond. 4. Overall the liquid component was more inhibited than the powder component of glass-ionomer cement in SDH activity test. 5. In $^{51}Cr$-release test, Fuji lining LC was the most released of all the cements tested and followed by : Vitrebond, Ketac-bond, GC lining cement. 6. In viable cell count, the number of cells increased as the culture day proceeded in Ketac-bond, but they decreased in GC lining cement. Fuji lining LC was only observed after 2 days culture and there was not observed the whole culture days in Vitrebond. 7. In $^{14}C$-leucine incorporation rate test, protein synthesis was decreased with the number of culture days in GC lining cement, Vitrebond and Fuji lining LC, but it was followed that of control in Ketacbond.

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Volatile Flavor Components in Mash of Takju prepared by using Aspergillus oryzae Nuruks. (Aspergillus oryzae 누룩으로 담금한 탁주 술덧의 발효 과정 중 휘발성 향기성분)

  • Lee, Taik-Soo;Han, Eun-Hey
    • Korean Journal of Food Science and Technology
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    • v.33 no.3
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    • pp.366-372
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    • 2001
  • Volatile flavor components in the mash of Takjus prepared by using Aspergillus oryzae nuruk were identified by using Gas Chromatography and Gas Chromatography-Mass Spectrometry. Twenty-four esters, 21 alcohols, 10 acids, 9 aldehydes and 4 others were found in the mash of Takju. Thirty six components including 13 esters and 12 alcohols were detected in the beginning of fermentation. Twenty nine components were more detected after second day of fermentation and 68 components were detected after 12 days of fermentation. Thirty five flavor components including 12 alcohols such as ethanol, 2-methyl-1-propanol, 3-methyl-1-butanol and benzeneethanol, 13 esters such as ethyl acetate, ethyl caprylate, ethyl butyrate and isoamyl acetate, 4 aldehydes and 6 acids were usually detected in the fermentation process. Ethanol was predominantly found in the range of $79.86{\sim}89.54%$ as a major component by using relative peak area. 3-Methyl-1-butanol, ethyl caprylate and benzeneethanol were some of the major volatile components through the fermentation respectively. Peak area of 2-methyl-1-propanol, 1-hexanol, 1-dodecanol, ethyl acetate, monoethyl butanoate, acetic acid and isobutylaldehyde among the same group were higher than other components depending upon fermentation time.

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Volatile Flavor Components in Mash of Takju prepared by using Aspergillus kawachii Nuruks (Aspergillus kawachii 누룩으로 담금한 탁주 술덧의 발효 과정 중 휘발성 향기성분)

  • Lee, Taik-Soo;Choi, Jin-Young
    • Korean Journal of Food Science and Technology
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    • v.37 no.6
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    • pp.944-950
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    • 2005
  • Volatile flavor components of Takjus mash prepared using Aspergillus kawachii nuruk were identified by GC and GC/MS. Twenty-two esters, 20 alcohols, 10 acids, 8 aldehydes, and 3 others were found in Takju mash. Thirty two components including 13 esters and 13 alcohols were detected at beginning of fermentation. Thirteen more components were detected after second day of fermentation, and 63 additional components after 12 days of fermentation. Twenty nine flavor components including 12 alcohols such as ethanol, 3-methyl-1-butanol, 2-methyl-1-propanol, and benzeneethanol, 12 esters such as ethyl acetate, ethyl caprylate, and ethyl butyrate 3 aldehydes, and 2 acids were detected during fermentation. Major volatile components detected during fermentation included 3-methyl-1-butanol, ethyl caprylate, and benzeneethanol. Peak areas of 2-methyl-1-propanol, 1-hexanol, 2, 3-butanediol (D.L), 1-dodecanol, 2-phenylethyl acetate, ethyl acetate, and monoethyl butanoate were higher than those of other components depending upon fermentation period.

Development of L-Lysine Producing Strains from Cellulosic Substrate by the Intergeneric Protoplast Fusion- Conditions for Formation and Regeneration of Protoplast - (속간 원형질체 융합에 의한 섬유질 기질로부터 L-lysine 생산균주 개발 -원형질체의 형성 및 재생 -)

  • 성낙계;정덕화;이무영;정영철
    • Microbiology and Biotechnology Letters
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    • v.16 no.2
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    • pp.150-155
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    • 1988
  • In order to produce L-lysine from cellulosic substrates by the intergeneric protoplast fusion between cellulolytic bacteria, Cellulomonas flavigena KFCC31221 and amino acid producing bacteria, Brevibacterium flavum ATCC14067, Corynebacteriurn glutamicum ATCC13032, conditions for protoplast formation and regeneration of these strains were investigated. After the strains were mutated with 500$\mu\textrm{g}$/$m\ell$ N-methyl-N'-nitro N-nitrosoguanidine for 30 min and the mutants were enriched by treating 300$\mu\textrm{g}$/$m\ell$ penicillin-G for 2 hrs, B. flavum Hse- Str$^{r}$ , C. glutamicum Met$^{-}$Thr$^{-}$ Rif$^{r}$ and Cellulomonas flavigena Thr$^{-}$Val$^{-}$Kan$^{r}$ were isolated. The rate of protoplast formation ranged from 95 to 98% when strains were treated at the concentration of 500$\mu\textrm{g}$/$m\ell$ of lysozyme, pH 6.5, 33$^{\circ}C$, for 6 hrs. in Tris- malate buffer supplemented with 0.4M sucrose as osmotic stabilizer. Approximately 30-33% protoplast was regenerated on the regeneration complete medium(RCM) containing 1.5% agar and 0.5M sodium succinate overlaid with the same medium except 0.7% agar.

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Purification of Heat-Stable Enterotoxin of Enterotoxigenic Escherichia coli eKT-53 (장독성 대장균 eKT-53 균주의 내열성 장독소 정제)

  • Do, Dea-Hong;Kim, Kyo-Chang;Kim, Do-Young
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.21 no.1
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    • pp.76-83
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    • 1992
  • Enterotoxigenic E. coli is one of the major causative agents of the infantile diarrhea and traveler's diarrhea. The heat-stable enterotoxin (ST) is thought to be a virulence factor in the pathogenesis of the diarrhea and to be a maker for identification of the enterotoxigenic E. coli from non pathogenic E. coli. ST producing E. coli KM-7 strain was isolated from the swine and molecular cloning of ST gene of KM-7 strain. Transformant eKT-53 $(ST^+,\;LT^-)$ was selected by infant mouse assay (IMA). The culture supernatant of eKT-53 strain was performed purification by multipled steps. The culture supernatant (crude ST) was purified by sequentially applying batch adsorption chromatography on Amberlite XAD-2 resin, ion exchange chromatography on DEAE-Sephacel anion exchanger, gel filtration chromatography on Bio-Gel P-6 and preparative polyacrylamide slab gel electrophoresis. About 113-fold purification was achieved with a yield of about 11% of crude ST and the minimum effective dose(MED) of this purified ST was about 2.8ng in IMA. Homogeneity of purified ST was demonstrated by showing a single band in analytical SDS polyacrylamide disc gel electrophoresis.

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Extraction Equilibria of Succinic Acid by Using Aqueous Two Phases System Containing Imidazolium Ionic Liquids and Salts (이미다졸계 이온성액체와 염을 포함한 수상이성분계를 이용한 숙신산의 추출 평형)

  • Lee, Yong Hwa;Kang, Jeong Won;Hong, Yeon Ki;Kim, Ki-Sub
    • Clean Technology
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    • v.20 no.4
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    • pp.349-353
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    • 2014
  • Succinic acid is an important precursor in industries producing biopolymers, pharmaceutical and food additives and green solvents. However, due to the high price of petroleum and the global $CO_2$ emission, the biological production of succinic acid from renewable biomass is a novel process due to the fixation of $CO_2$ into succinate during fermentation. In this study, aqueous two phase systems based on imidazolium ionic liquids/$K_2HPO_4$ were used as an effective separation and concentration process for succinic acid. Experimental results show that aqueous two phase systems can be formed by adding appropriate amount of imidazolium ionic liquids to aqueous $K_2HPO_4$ solutions in the presence of succinic acid. It can be found that the ability of imidazolium ionic liquids for phase separation followed the order [HMIm][Br]${\fallingdotseq}$[OMIm][Br]>[BMIm][Br]>[EMIm][Br]. The maximum value of extraction efficiency for succinic acid was about 90% and the amount of coextracted water into top phase is proportional to the chain length of cation in imidazolium ionic liquids. It was concluded that the aqueous two phase systems composed of imidazolium ionic liquids/$K_2HPO_4$ was effective for the selective extraction and concentration of succinic acid.

Chemical composition changes in fermented Doenjang depend on Doenjang koji and its mixture (된장 koji 및 그 혼합에 따른 된장 숙성 과정중의 화학성분 변화)

  • Joo, Hyun-Kyu;Kim, Dong-Hyun;Oh, Kyun-Teak
    • Applied Biological Chemistry
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    • v.35 no.5
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    • pp.351-360
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    • 1992
  • In order to improve the qualities of Doenjang were investigated on the enzyme activity in the koji and changes in chemical composition, flavors and sensory envaluation of Doenjang which were prepared with Rh. delemar koji, Asp. oryzae koji and traditional Meju with mixed koji and soybean as the ratio of optimum mixture. Asp. oryzae koji was indicated highest activities ${\alpha}$ and ${\beta}-amylase$ as 312 mg/ml, 235 mg/ml while Rh. delemar the lowest activities as 16 mg/ml, 38 mg/ml in aging for 40 days amino type ntrogen was the highest in the Asp. oryzae and Rh. delmear mixture koji(D group), Asp. oryzae(A group), and Asp. oryzae koji and Rh. delemar koji(C group) as 460 mg%, 440 mg% and 426 mg% in aging for 40 days. The main flavor components of Doenjang were detected as fellows phenol-2-kmethoxy, 4H-pyran-4-one-3-hydroxy-2-methyl, benzenthanol, 1-octan-3-ol, tetra-methyl pryrazine, 1,3,6 cyclooctatrien. Asp. oryzae(A) and Asp. oryzae koji with Rh. delemar koji mixture(C), group were the most excellent in taste, flavor color for fermented Doenjang at 40 days.

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Preparation Conditions of Agar Oligosaccharides with Organic Acids (한천 올리고당 제조를 위한 유기산 처리 조건)

  • JOO Dong-Sik;KIM Ok-Seon;CHO Soon-Yeoung;LEE Chang-Ho
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.36 no.1
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    • pp.6-10
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    • 2003
  • The optimum organic acid and temperature conditions were investigated for the preparation of oligosaccharides from agar. The tested organic acids were acetate, citrate, lactate, malate, and succinate and the conditions for oligosaccharides preparation were $0.3\%,\;0.5%;and\;0.7\%$ organic acid concentrations at $80\~120^{\circ}C.$ The low concentration of organic acid below $0.3\%$ decreased the degrading ratio and the high concentration up $0.5\%$ could not changed the degrading ratio. Conditions below $100^{\circ}C$ was not good for degrading agar. But $100^{\circ}C\;or\;120^{\circ}C$ was optimal temperature conditions for agarooligosaccharides according to the organic acid type and concentration. The organic acid concentration was $0.5\%$ and organic acid was the citrate or malate. The treatment time considered optimum was 120$\~$180 min. The maximal degrading ratio giving optimum conditions such as $100^{\circ}C\;and\;120^{\circ}C\;was\;35.5\%\;and\;38.7\%,$ respectively. The agarooligosaccharides prepared by autoclaving at $100^{\circ}C\;and\;120^{\circ}C$ were 2$\~$7 species oligomer.