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Development of an Analytical Method for Fluxapyroxad Determination in Agricultural Commodities by HPLC-UVD (HPLC-UVD를 이용한 농산물 중 Fluxapyroxad 잔류분석법 개발)

  • Kwon, Ji-Eun;Kim, HeeJung;Do, Jung-Ah;Park, Hyejin;Yoon, Ji-Young;Lee, Ji-Young;Chang, Moon-Ik;Rhee, Gyu-Seek
    • Journal of Food Hygiene and Safety
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    • v.29 no.3
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    • pp.234-240
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    • 2014
  • Fluxapyroxad is classified as carboxamide fungicide that inhibits succinate dehydrogenase in complex II of mitochondrial respiratory chain, which results in inhibition of mycelial growth within the fungus target species. This study was carried out to assure the safety of fluxapyroxad residues in agricultural products by developing an official analytical method. A new, reliable analytical method was developed and validated using High Performance liquid Chromatograph-UV/visible detector (HPLC-UVD) for the determination of fluxapyroxad residues. The fluxapyroxad residues in samples were extracted with acetonitrile, partitioned with dichloromethane, and then purified with silica solid phase extraction (SPE) cartridge. Correlation coefficient($R^2$) of fluxapyroxad standard solution was 0.9999. The method was validated using apple, pear, peanut, pepper, hulled rice, potato, and soybean spiked with fluxapyroxad at 0.05 and 0.5 mg/kg. Average recoveries were 80.6~114.0% with relative standard deviation less than 10%, and limit of detection (LOD) and limit of quantification (LOQ) were 0.01 and 0.05 mg/kg, respectively. All validation parameters were followed with Codex guideline (CAC/GL 40). LC-MS (Liquid Chromatograph-Mass Spectrometer) was also applied to confirm the analytical method. Base on these results, this method was found to be appropriate fluxapyroxad residue determination and can be used as the official method of analysis.

Formaldehyde Monitoring of Hygiene Products in Domestic Market (국내 유통 위생용품 중 포름알데히드 잔류량 모니터링)

  • Na, Young-Ran;Kwon, Hyeon-Jeong;Cho, Hyun-Nho;Kim, Hyeon-Jin;Park, Yon-Koung;Park, Sung-Ah;Lee, Seong-Ju;Kang, Jung-Mi
    • Journal of Food Hygiene and Safety
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    • v.35 no.3
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    • pp.225-233
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    • 2020
  • By the standards and specifications for hygiene products, three test methods for formaldehyde are specified for each item type of hygiene product. After derivatization using acetylacetone and 2,4-dinitrophenylhydrazine (2,4-DNPH), formaldehyde is analyzed by spectrophotometer and high-performance liquid chromatography (HPLC). Validation of the three test methods was performed on tissue, diaper lining and waterproof layer, and panty liner products. The results of linearity (R2), limit of detection (LOD), limit of quantification (LOQ), recovery rate (%) and reproducibility (%), showed that all three methods are suitable for analyzing formaldehyde in hygiene products. After derivatization with 2,4-DNPH and cetylacetone, formaldehyde was analyzed at 0, 3, 6, 9, 24 and 48 hours by HPLC. Formaldehyde derivatized with 2,4-DNPH showed no statistically significant change in formaldehyde peak area over time (P>0.05). But, acetylacetone-derivatizated formaldehyde showed a negative correlation coefficient (r) over time (P<0.01). We investigated the residual amounts of formaldehyde in 205 hygiene products distributed in Busan. Among 74 disposable diaper products tested, 73 had low concentrations of formaldehyde (0.13-29.87 mg/kg). Moreover, formaldehyde was not detected in any of 78 tissue, 27 disposable paper towel, 12 disposable dishcloth, 7 paper cup, one brand of paper straw and 6 disposable napkin products.

Recent Advancement in the Differentiation of Tissues and Organs and Regulation of Gene Expression (조직.기관의 분화와 유전자 발현의 조절, 최근의 진보)

  • Harn, Chang-Yawl
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.1
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    • pp.1-35
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    • 1997
  • Fertilized egg, by successive cell divisions, differentiates into different tissues and organs with various structures and functions. Different cells and tissues contain different proteins, products of selective gene expression. Not all the genes in any genomes are equally active, temporal and spatial gene expression being the general rule. Present paper attempts to review the tanscriptional mechanisms or the initiations of transcription from several angles. In some of the organisms the genes in the process of transcription or the genes in the inactive state can be seen under the light microscope. Some bands of Drosophila polytene chromosomes may exhibit a swollen or puff appearance under certain conditions. A puff, unfolded or decondensed form of chromomere, represents sets of intense transcriptional activity or RNA synthesis. The heterochromatic X chromosome whose genes remain inactive in the female mammals can be visualized as a dark staining structure called Barr body, Configuration of chromatin differs between transcribed and nontranscribed chromatin. Modification to the chromatin facilitates RNA synthesis. The movement of large polymerase molecule along the DNA would probably be facilitated if some modifications of the chromatin configuration is effected. Methylation of cytosines in CG sequences is associated with inactive genes. Methylation can play a role in determination of mammalian cells during embryogenesis. Demethylation is necessary for the gene to be expressed during development A histone modification that is also known to be correlated with transcriptional capacity of chromatin is acetylation of the lysine residues of the core histones. Chromatin containing a high level of histone acetylation is very sensitive to DNase 1. For the transcription to occur TBP must first bind to the TATA box. Another TF, TF IIB, then binds to the promoter-TBP complex, facilitating the access of RNA polymerase to the transcription initiation site. As recently as eight years ago researchers assumed that histones were irrelevant to the regulation of gene expression. Histones combine with the DNA to form nucleosome of the chromatin. Histones are vital participant in gene regulation. Histone and basal factors compete for access to TATA box. When DNA is exposed to basal factors before histones are introduced, the basal factors assemble on TATA boxes preventing the access of histones, allowing transcription to occur, for transcription to begin, activator protein at the upstream activation sequence or enhancer must interact with the tail of histone H4 at TATA box and cause the histone role particle to dissociate from the TATA box leading to partial breakup of the histone core particle and allowing the basal factors to bind to the TATA box. New concept of genomic flux in contrast to the old concept of static genome has been developed based on the powerful new molecular techniques. Genomic changes such as repetitive DNAs and transposable elements, it is assumed but not yet proved, may affect some of the developmental patterns that characterize particular cells, tissues, organs, and organisms. In the last decade or so remarkable achievement have been made in the researches of the structures and functions of TFs and the specific target sequences located in promoters or enhancers where these TFs bind. TFs have independent domains that bind DNA and that activate transcription. DNA binding domain of TFs serves to bring the protein into the right location. There are many types of DNA binding domains. Common types of motifs can be found that are responsible for binding to DNA. The motifs are usually quite short and comprise only a small part of the protein structure. Steroid receptors have domains for hormone binding, DNA binding, and activating transcription. The zinc finger motif comprises a DNA binding domain. Leucine zipper consist of a stretch of amino acids with a leucine residue in every seventh position Two proteins form a dimer because they interact by means of leucine zippers on similar α-helical domain. This positions their DNA binding basic domains for interaction with the two halves of a DNA sequence with dyad symmetry of TGACTCA, ACTGAGT.

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Investigation on Inhibitory Effect of ErmSF N-Terminal End Region Peptide on ErmSF Methyltansferase Activity In Vivo Through Development of Co-Expression System of Two Different Proteins in One Cell (서로 다른 두 단백질의 세포 내 동시 발현 체계의 개발을 통한 ErmSF에서 특이적으로 발견되는 N-Terminal End Region (NTER)을 포함하는 펩타이드의 생체내에서의 ErmSF 활성 억제 효과 검색)

  • Jin, Hyung-Jong
    • Korean Journal of Microbiology
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    • v.47 no.3
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    • pp.200-208
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    • 2011
  • Most problematic antibiotic resistance mechanism for MLS (macrolide-lincosamide-streptogramn B) antibiotics encountered in clinical practice is mono- or dimethylation of specific adenine residue at 2058 (E. coli coordinate) of 23S rRNA which is performed by Erm (erythromycin ribosome resistance) protein through which bacterial ribosomes reduce the affinity to the antibiotics and become resistant to them. ErmSF is one of the four gene products produced by Streptomyces fradiae to be resistant to its own antibiotic, tylosin. Unlike other Erm proteins, ErmSF harbors idiosyncratic long N-terminal end region (NTER) 25% of which is comprised of arginine well known to interact with RNA. Furthermore, NTER was found to be important because when it was truncated, most of the enzyme activity was lost. Based on these facts, capability of NTER peptide to inhibit the enzymatic activity of ErmSF was sought. For this, expression system for two different proteins to be expressed in one cell was developed. In this system, two plasmids, pET23b and pACYC184 have unique replication origins to be compatible with each other in a cell. And expression system harboring promoter, ribosome binding site and transcription termination signal is identical but disparate amount of protein could be expressed according to the copy number of each vector, 15 for pACYC and 40 for pET23b. Expression of NTER peptide in pET23b together with ErmSF in pACYC 184 in E. coli successfully gave more amounts of NTER than ErmSF but no inhibitory effects were observed suggesting that there should be dynamicity in interaction between ErmSF and rRNA rather than simple and fixed binding to each other in methylation of 23S rRNA by ErmSF.

Monitoring of Pesticide Residues in Leafy Vegetables Collected from Wholesale and Traditional Markets in Cheongju (청주지역 도매 및 재래시장 유통 엽채류 중 잔류농약 모니터링)

  • Noh, Hyun-Ho;Park, Young-Soon;Kang, Kyung-Won;Paik, Hyo-Kyung;Lee, Kwang-Hun;Lee, Jae-Yun;Yeop, Kyung-Won;Choi, Song-Rim;Kyung, Kee-Sung
    • The Korean Journal of Pesticide Science
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    • v.14 no.4
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    • pp.381-393
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    • 2010
  • In order to monitor the residual characteristics of the pesticides in leafy vegetables selling at wholesale markets and traditional markets in Cheongju, a total of 180 samples of 15 leafy vegetables, such as broccoli, celery, chard, chicory, Chinese vegetable, Chwinamul, crown daisy, Korean cabbage, leek, lettuce, perilla leaves, Shinsuncho, spinach, welsh onion and young radish, were purchased from the wholesale markets and traditional markets in June and August in 2010 and the pesticide residues in them were analyzed by multiresidue analysis method using GLC, HPLC and GC-MSD. Seven pesticides were detected from 12 samples out of total 180 samples collected, representing detection rate was 6.7%. In case of the samples collected from markets in June, four pesticides including tefluthrin were detected from six samples and in case of the samples collected from markets in August, three pesticides including pendimethalin were detected from three samples. The MRL-exceeding rate of pesticides detected from leafy vegetables was 0.6%. The pesticide exceeded its MRL was azoxystrobin detected from crown daisy and many pesticides were not registered to the crops, excepting that azoxystrobin detected from Chwinamul and tefluthrin from leek. Estimated daily intakes (EDIs) of the pesticides detected from leafy vegetables were less than 7% of their acceptable daily intakes (ADIs), representing that residue levels of the pesticides detected were evaluated as safe.

Biological Control of the Mushroom Fly, Lycoriella mali, Using Bacillus thuringiensis subsp. israelensis (Bacillus thuringiensis subsp. israelensis를 이용한 느타리 재배에서 버섯파리의 생물학적 방제)

  • Moon, Byung-Ju;Lee, Su-Hee;Lim, Eun-Kyung;Kim, Tae-Sung;Kim, Hyun-Ju;Song, Ju-Hee;Kim, Ik-Soo
    • The Korean Journal of Mycology
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    • v.30 no.1
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    • pp.50-55
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    • 2002
  • Biological control against mushroom fly, Lycoriella mali, was performed by using Bacillus thuringiensis subsp. israelensis Bti-D and Bti-U, isolated from dead mushroom fly in oyster mushroom houses. Control values of the bacterial strains Bti-D and Bti-U against L. mali in bottle culture of oyster mushroom were 74.4% and 64.2%, respectively, and the value in small tray culture were 75.8% and 56.8%, respectively. In the experiment to develop the mass, cheap media for Bti-D and Bti-U isolates, the Biji broth (bean curd residue, called Biji in Korean language) was selected as a culture medium for an inexpensive and mass cultivation by the measurement of optical density of the two bacteria grown in the different media tested. Insecticidal effect of the formulation contained different ingredients that were prepared by using the Bti-D strain cultured in the Biji broth was tested in tray and bottle culture of oyster mushroom. The WCS formulation that contained corn starch as bio-gel (86.4%) was more effective to control the mushroom fly than living cells (69.1%) in bottle culture of oyster mushroom. Moreover, insecticidal effect of the WCS formulation was improved when water of pH 8 was used for dilution of the formulation. Effect of the WCS formulation using water of pH 8 and chemicals, Zuron (dimillin) W.P. on the control of mushroom fly and the productivity of oyster mushroom was investigated in tray culture of oyster mushroom. The Zuron W.P. was more effective to control the mushroom fly than the WCS formulation. However, compared with no treatment, the productivity of the mushroom treated with the WCS formulation was improved than that of the mushroom with Zuron W.P.

Effect of Low Concentrated-phosphorous Fused Phosphate on Rice Plant (수도(水稻)에 대(對)한 인산저농도(燐酸低濃度) 용성인비(鎔成燐肥)의 효과(效果))

  • Lee, Yun Hwan;Han, Ki Hak;Park, Young Dae;Kim, Bok Jin;Heu, Ii Bong
    • Korean Journal of Soil Science and Fertilizer
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    • v.5 no.1
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    • pp.1-8
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    • 1972
  • In order to expect the effect of silica with large quantity application of current Fused calcium-magnesium phosphate on the paddy rice, there are difficulties of excess phosphorus application because of the high content of phosphate in this fertilizer. This experiment was discussed on the effect of posphate and silica absorbed by rice plant from the low concentrated fused calcium-magnesium phosphate which was fused with mixture of rock phosphate, chemical calcium oxide, magnesium oxide and silicate oxide in the furnace using coke, 1. The fusion material contained 8.9% of citric acid soluble $P_2O_5$ and 33% of soluble $SiO_2$. 2. The rice yields were increased with high significance accompanying the application levels of fused material amounts. 3. No. of grains per head, weight of 1,000 grains and percent of filled grain were caused to increase the productivity of rice plant on account of the high content of silica in straws absorbed from fusion material. The treatment of 300 kg/10a. was the highest yield among the levels of fusion material. 4. At the growing periods of rice plant, amount of absorbed phosphate was higher in the small amount treatment of fusion material until the formation period of young head, and was highest in the treatment of 300 kg/10a. leval among them but slightly desreased at 500 kg/10a. level at the harvest. Amount of absorbed silica was the same trend with phosphorus at the begining of growth period but increased rapidly from the formation period of young head to harvest in the large quantity application levels. 5. Much amount of nutrients were residued in the soil after experiment pacing with application levels. 6. The effect of silica and phosphate on rice plant can be expected with fusion material but it is necessary to decrease the phosphate content on account of the large residue of phosphate in the soil after experiment.

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Quality Characteristics of Bijijang in Different Fermentation Conditions (발효 조건을 달리한 비지장의 품질특성)

  • Im, Sung-Kyung;Yoo, Seon-Mi;Kim, Tae-Young;Chun, Hye-Kyung
    • Korean Journal of Food Science and Technology
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    • v.36 no.3
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    • pp.448-455
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    • 2004
  • Changes in quality characteristics of Bijijang (fermented soybean curd residus) prepared at $35^{\circ}C\;and\;40^{\circ}C$ for 0, 12, 24, 36, and 48 hr were investigated. Acidity of Bijijang increased, whereas pH and Hunter's color values decreased during fermentation. Immediately after Bijijang preparation, ${\alpha}-and\;{\beta}-amylase$ activities were very low, ${\beta}-Amylase$ activity during fermentation increased rapidly, with those fermented at $40^{\circ}C$ higher than at $35^{\circ}C$. Neutral pretense activity was significantly higher than acidic pretense activity, and increased gradually after 12 hr. Change in total nitrogen content in Bijijang was insignificant, whereas contents of amino-type and water-soluble nitrogens increased significantly during fermentation. Major free amino acids of Bijijang were Arg, Pro, Glu, Thr, Ser, and Lys at initial fermenting stage, and, as fermentation progressed, contents of Cys, Met Glu, Ile, Leu, and Phe increased. Reducing sugar contents of Bijijang fermented at $40^{\circ}C$ were higher than those fermented at $35^{\circ}C$. Sucrose content decreased and glucose content increased. Glucoside (genistin and daidzin) contents decreased and aglycone (genistein and daidzein) contents increased during preparation of Biji and fermentation of Bijijang. Contents of free sugars and isoflavones were higher in Bijijang fermented at $40^{\circ}C$ than at $35^{\circ}C$. Based on these results, fermentation at $40^{\circ}C$ for 48 hr was determined to be optimum fermentation condition for Bijijang.

Studies on $\alpha$-amylase of Bocillus circulans F-2 (Part II) Enzymatic characteristics of the purified $\alpha$-amylase (Bacillus circulans F-2가 생산하는 $\alpha$-amylase에 관한 연구 (제 I I 보) 정제$\alpha$-amylase의 효소적특성)

  • ;Hajime Taniguchi;Yoshiharu Maruyama
    • Microbiology and Biotechnology Letters
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    • v.10 no.2
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    • pp.123-132
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    • 1982
  • These experiments were conducted to investigate the enzymatic characteristics of the purified $\alpha$-amylase (F-2A) of Bacillus circulans F-2 and the digestion rate of various starches. 1. The molecular weight was estimated to be 93000 by SDS-polyacrylamide disc gel electrophoresis. The isoelectric point was about pH 5.0. The optimum pH for the enzyme action was 6.0-6.5 and the stable pH ranged pH 5.5-12.0. The optimum temperature was 6$0^{\circ}C$, and the purified $\alpha$-amylase was stable below 4$0^{\circ}C$. 2. The purified $\alpha$-amylase was activated by Mn$^{++}$ and Co$^{++}$, whereas it was inhibited by Ag$^{+}$, HT$^{++}$, Cu$^{++}$ and Pb$^{++}$. 3. The purified $\alpha$-amylase is considered to have no sulfhydryl residue essential for its catalytic activity. 4. Michaelis constant (Km) was 1.704 mg/$m\ell$. Activation energy between 25-4$0^{\circ}C$ was 12.297 Kcal/mole, and between 40-6$0^{\circ}C$, it was 7.831 Kcal/mole. 5. The hydrolysis product from soluble starch, amylose and amylopectin in the early stage of hydrolysis was G$_{6}$, and as hydrolysis proceeds, G$_4$and G$_2$appeared. 6. Products from each oligosaccarides are as follows: G$_4$longrightarrow G$_2$+ G$_2$,G$_3$ +G$_1$,G$_{5}$longrightarrow G$_4$+G$_1$,G$_{6}$longrightarrowG$_4$+ G$_2$,G$_{7}$ G$_4$,G$_{8}$longrightarrow G$_4$+G$_4$, 7. On raw potato starch, raw sago starch and raw yam starch, the purified enzyme exhibited a remarkably high digestion rate than Porcine pancreatic amylase and Streptococcus bovis amylase.

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Antiviral Activity of Hot-Water Extract and Its Ethanol Precipitate of Phellinus pini Fruiting Body (낙엽진흙버섯 자실체 유래 열수 추출물과 메탄을 침전물의 항바이러스활성)

  • Lee, Sae-Mi;Kim, Sung-Min;Lee, Yoon-Hee;Kim, Woo-Jung;Na, Ye-Seul;Kim, Hyun-Guell;Nam, Jae-Hwan;Shin, Hyun-Dong;Kwon, Doo-Han;Park, Yong-Il
    • Microbiology and Biotechnology Letters
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    • v.37 no.1
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    • pp.33-41
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    • 2009
  • The crushed fruiting body of Alaskan Porodaedalea pini (Brot.) Murrill (syn. Phellinus pini) was extracted in boiling water for 4 h with the yield of 20.5% in dry mass. This hot-water extract showed significant antiviral activity by inhibiting the plaque formation in HeLa cells by coxackievirus B3 (CVB3) and also showed highest inhibitory effect against neuraminidase activity among water extracts of various mushrooms. From the water extract, the ethanol precipitate (EP) and supernatant fraction (ES) were obtained through 75% ethanol precipitation with the yield of 43.3% and 28.3% in dry mass, respectively. Whereas ES did not show any detectable level of antiviral activity, EP showed significant dose-dependent inhibition of plaque formation by CVB3 in HeLa cells with an $EC_{50}$ (50% effective concentration) of 0.45 mg/mL. The cytotoxicity on HeLa cells by EP was relatively low with the $CC_{50}$ (50% cytotoxic concentration) of 2.25 mg/mL. EP also effectively inhibited neuraminidase activity in a dose-dependent manner showing up to 75% inhibition at 1.7 mg/mL. These results suggest that the hot-water extract and its EP of P. pini fruiting body can be a candidate for the development of a potent broad-range antiviral agent against influenza virus(Flu) as well as CVB3. The major active component of EP was shown to be a heteropolysaccharide-protein complex containing glucose as the main sugar residue with mole percentage of 79.8% and other sugars like galactose (19.2%), xylose (17%), mannose (5.8%), and fucose (4.6%) and a small portion (12.7%, in mass) of protein.