• Title/Summary/Keyword: Gel electrophoresis

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Biological Activity of Fermented Silkworm Powder (발효누에분말의 생리활성)

  • Cha, Jae-Young;Kim, Yong-Soon;Ahn, Hee-Young;Eom, Kyung-Eun;Park, Bo-Kyung;Jun, Bang-Sil;Cho, Young-Su
    • Journal of Life Science
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    • v.19 no.10
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    • pp.1468-1477
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    • 2009
  • The comparative effects of the fibrinolytic, and tyrosinase inhibition activities and electrophoretical protein patterns with freeze-drying silkworm powder (FDSW), heating-drying silkworm powder (HDSW) and fermented silkworm powder by Bacillus subtilis or Lactobacillus hilgardii were investigated. When total protein patterns of FDSW, HDSW, both fermented SW, were analyzed by native- and SDS-polyacrylamide gel electrophoresis (PAGE), there were slightly varietal differences in electrophoretical protein patterns. Major minerals of FDSW and HDSW were K, Ca, Mg, and Zn. Major compositional amino acids of FDSW and HDSW were glycine, alanine, glutamic acid, aspartic acid, and serine. Major fatty acids of FDSW and HDSW were linolenic acid, oleic acid, and palmitic acid. Fibriolytic activity was the highest in the fermented FDSW by 5% B. subtilis among the various samples. Tyrosinase inhibition activity was higher in the water and 70% methanolic extract of FDSW than in HDSW. DPPH radical scavenging activity was slightly stronger in HDSW than in FDSW. In addition, DPPH radical scavenging activity was higher in FDSW or HDSW fermented by L. hilgardii than that fermented by B. subtilis, however, all samples exhibited a relatively low activity compared to the butylated hydroxytoluene (BHT). These results may provide the basic data to understand the biological activities of fermented SW.

Isolation and Characterization of Bacillus licheniformis SC082 Degrading Fibrin and Chitin from Shrimp Jeot-Gal (새우젓으로부터 혈전과 chitin 분해능을 지닌 균주 Bacillus licheniformis SC082의 분리 및 특성)

  • Cho, Eun-Kyung;Jung, Yu-Jung;Gal, Sang-Wan;Choi, Young-Ju
    • Journal of Life Science
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    • v.19 no.10
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    • pp.1424-1431
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    • 2009
  • Shrimp Jeot-Gal is a popular traditional Korean fermented seafood and has been used for seasoning. We isolated a bacterium showing strong extra-cellular fibrinolysis and chitinase activity from shrimp Jeot-Gal and the strain was designated SC082. SC082 was identified as Bacillus licheniformis by 16S rRNA sequence homology search. B. licheniformis SC082 exhibited optimum temperature, pH, and salt concentration at $37^{\circ}C$, pH 7.0, and 6%, respectively. Substrate specificity of the culture supernatant from B. licheniformis SC082 was detected in fibrin, skim milk, and chitin plate. The fibrinolytic activity was highly maintained up to $50^{\circ}C$ at a pH of 7.0 for 3 hr and was stable up to pH 9.0 at $37^{\circ}C$ for 3 hr. The chitinase activity was remarkably induced by addition of 1.0% colloidal chitin and the pH and temperature optima of the enzyme were 5.0 and $45^{\circ}C$, respectively. In sodium dodecyl sulfate-polyacrylamide gel electrophoresis and zymogram analysis, this strain produced three fibrinolytic isozymes and two chitinase isozymes. The approximate molecular weights of the putative fibrinolytic enzymes were 23.0, 62.0, and 72.0 kDa and those of the chitinases were 62.0 and 55.0 kDa, respectively. The antioxidant activity of SC082 was also measured by using 2,2-diphenyl-l-picryl-hydrazyl (DPPH) free radical. The DPPH radical scavenging was slightly increased in a dose-dependent manner.

Serum Anti-Gal-3 Autoantibody is a Predictive Marker of the Efficacy of Platinum-Based Chemotherapy against Pulmonary Adenocarcinoma

  • Yanagita, Kengo;Nagashio, Ryo;Ryuge, Shinichiro;Katono, Ken;Jiang, Shi-Xu;Tsuchiya, Benio;Nakashima, Hiroyasu;Fukuda, Eriko;Goshima, Naoki;Saegusa, Makoto;Satoh, Yukitoshi;Masuda, Noriyuki;Sato, Yuichi
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.17
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    • pp.7959-7965
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    • 2015
  • Background: Identification of predictive markers for the efficacy of platinum-based chemotherapy is necessary to improve the quality of the life of cancer patients. Materials and Methods: We detected proteins recognized by autoantibodies in pretreated sera from patients with lung adenocarcinoma (AC) evaluated as showing progressive disease (PD) or a partial response (PR) after cisplatin-based chemotherapy by proteomic analysis. Then, the levels of the candidate autoantibodies in the pretreated serum were validated by dot-blot analysis for 22 AC patients who received platinum-based chemotherapy, and the expression of identified proteins was immunohistochemically analyzed in 40 AC biopsy specimens. Results: An autoantibody against galectin-3 (Gal-3) was detected in pretreated sera from an AC patient with PD. Serum IgG levels of anti-Gal-3 autoantibody were significantly higher in patients evaluated with PD than in those with PR and stable disease (SD) (p = 0.0084). Furthermore, pretreated biopsy specimens taken from patients evaluated as showing PD following platinumbased chemotherapy showed a tendency to have a higher positive rate of Gal-3 than those with PR and SD (p = 0.0601). Conclusions: These results suggest that serum IgG levels of anti-Gal-3 autoantibody may be useful to predict the efficacy of platinum-based chemotherapy for patients with lung AC.

Crocetin Induces Cytotoxicity in Colon Cancer Cells Via p53-independent Mechanisms

  • Li, Cai-Yan;Huang, Wen-Feng;Wang, Qun-Li;Wang, Fan;Cai, E.;Hu, Bing;Du, Jia-Cheng;Wang, Jing;Chen, Rong;Cai, Xiao-Jing;Feng, Jing;Li, Hui-Hui
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.8
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    • pp.3757-3761
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    • 2012
  • Objective: Crocin has been proposed as a promising candidate for cancer chemoprevention. The purpose of this investigation was to investigate the chemopreventive action and the possible mechanisms of crocin against human colon cancer cells in vitro. Methods: Cell proliferation was examined using MTT assay and the cell cycle distribution fractions were analyzed using fow cytometric analysis after propidium iodide staining. Apoptosis was detected using theTUNEL Apoptosis Detection Kit with laser scanning confocal microscope. DNA damage was assessed using the alkaline single-cell gel electrophoresis assay, while expression levels of p53, cdk2, cyclinA and P21 were examined by Western blot analysis. Results: Treatment of SW480 cells with crocetin (0.2, 0.4, 0.8 mmol/L) for 48 h signifcantly inhibited their proliferation in a concentration-dependent manner. Crocetin (0.8 mmol/L) signifcantly induced cell cycle arrest through p53-independent mechanisms accompanied by P21 induction. Crocetin (0.8 mmol/L) caused cytotoxicity in the SW480 cells by enhancing apoptosis and decreasing DNA repair capacity in a time-dependent manner. Conclusions: This report provides evidence that crocetin is a potential anticancer agent, which may be used as a chemotherapeutic drug.

PEROXIDASE ACTIVITY ASSOCIATED WITH Puccinia sorghi INFECTION IN MAIZE (옥수수에 있어서 녹병감염에 관한 Peroxidase의 활성)

  • Kim Soon Kwon;Brewbaker James L.;Hasegawa Yoichi
    • Korean journal of applied entomology
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    • v.17 no.4 s.37
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    • pp.193-199
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    • 1978
  • Five parents and their 10 $F_1$ crosses of maize (Zea mays L.) were tested by means of diallel analysis for the inheritance of peroxidase activity following Puccinia sorghi rust infection. Peroxidase activity was measured at day zero and at 2 days and 8 days after inoculation. Peroxidase activity was increased significantly by P. sorghi infection in all 15 genotypes after 8 days, but not after 2 days. Highly significant differences in peroxidase activity were detected among the 15 genotypes. The highly genera] resistant inbred, CM105, and its hybrids showed exceptionally high peroxidase activity in both healthy and infected plants. However, another highly resistant inbred, Oh545, showed exceptionally low peroxidase activity. Significant general combining ability (GCA) and specific combining ability (SCA) means quares were detected for peroxidase activity independent of disease. GCA mean spuares, however, were consistently a major contiribution to the inheritance of peroxidase activity in the infected plants whereas SCA men square contributions were minor. Rust resistant maize plants controlling mongenic dominant $Rp_1^d$gene showed stronger peroxidase reroxidase responses than their susceptible counterparts in the gel electrophoresis and densiometric tracings. The increased peroxidase activity occurred in both major leaf peroxidases, $Px_3\;and\; Px_7$.

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Genetic variation and population structure of Asarum misandrum (Aristolochiaceae) in Korea (각시족도리풀(Asarum misandrum)의 유전적 다양성 및 집단 구조)

  • So, Soonku;Kim, Muyeol
    • Korean Journal of Plant Taxonomy
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    • v.43 no.3
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    • pp.181-187
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    • 2013
  • Genetic variation in Asaum misandrum, a woodland herb in Korea, was investigated based on allozyme analysis with starch gel electrophoresis. All of previously reported populations in Korea were sampled and seven loci from six enzymes were analyzed. Overall genetic variation of A. misandrum population showed considerably high levels of genetic variation within the species (A = 2.05, P = 71.4, $H_E$ = 0.294). A positive $F_{IS}$ value of A. misandrum indicated overall deficiency of heterozygotes, and a low $F_{ST}$ value (0.112) meant very little differentiation among populations. Factors contributing to the high levels of genetic diversity found within populations of A. misandrum include population maintenance via wide distribution range from Korea to Japan and primarily outcrossing breeding system. Although it showed moderate genetic diversity level, most habitats of the species were scattered and discontinuous. Besides, low numbers of individuals were found in the most habitats and individuals are collected frequently from the wild due to the unique shape of the flowers as well as the rarity of the species. Thus, there is a need to set up a reasonable conservation strategies including the maintenance mechanism of genetic diversity of A. misandrum.

Stabilization of Membrane Proteins by Benzyladenine during Wheat Leaf Senescence (노쇠중인 밀잎에서 Benzyladenine에 의한 막단백질의 안정화)

  • 진창덕
    • Journal of Plant Biology
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    • v.35 no.2
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    • pp.117-123
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    • 1992
  • The effect of benzyladenine (BA) on lipid peroxidation and compositions of total insoluble proteins and chloroplast thylakoid protein from wheat primary leaves during senescence in the dark was studied. BA ($10^{-5}\;M$) treatment prevented conspicuously the loss of chlorophyll content and soluble and insoluble leaf protein contents in senescing wheat leaf segments during 4-day dark incubation. Under the BA treatment, especially, the level of insoluble protein was highly maintained than that of soluble protein. Also, the increase of malondialdehyde (MDA: the peroxidation product of membrane lipids) content was inhibited in the BA treated leaves. Three major polypeptide bands in quantity corresponding to 57, 26 and 12 KD molecular weight were clearly resolved with other minor bands by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) in the insoluble protein fraction. The insoluble protein profiles of the control leaves showed a remarkable decrease in the intensity of the 57 and 12 KD band except for 26 KD band in the 72 h dark incubation. This loss during dark incubation was reduced by BA treatment. More than 20 polypeptides were resolved in the chloroplast thylakoid membrane fraction with the most prominent bands which are 59 and 57 KD ($\alpha\;and\;\beta$ subunit of coupling factor: CF) and 26 KD (apoprotein of LHCP). The changes in thylakoid protein profile during 72 h dark incubation showed the rapid degradation in control, but this degradation was prevented in quantity by BA treatment. The above results suggested that BA would inhibit the peroxidation of membrane lipids, thereby preventing the loss of membrane proteins which led to the maintenance of the membrane integrity including chloroplast thylakoid.

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Impact of phosphorus application on the indigenous arbuscular mycorrhizal fungi, soybean growth and yield in a 5-year phosphorus-unfertilized crop rotation

  • Higo, Masao;Sato, Ryohei;Serizawa, Ayu;Gunji, Kento;Suzuki, Daisuke;Isobe, Katsunori
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.351-351
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    • 2017
  • Arbuscular mycorrhizal fungi (AMF) are particular soil fungi that benefit many crops and require a symbiosis with plant roots to survive. In our previous study, there was a positive correlation between AMF root colonization and soybean grain yield in a four-year consecutive winter cover crop-soybean rotational system without phosphorus fertilizer. It is suggested that higher AMF root colonization can be a better solution for improving soybean growth and grain yield in P-limited soil. Our purpose in this study was to test the hypothesis that a P application is the main factor improving soybean growth, P nutrition and grain yield, and the benefit from AMF to soybean P uptake and growth in a P-limited soil. Impact of a P application on AMF root colonization and communities in soybean roots and their potential contribution to soybean growth and P nutrition under a five-year P-unfertilized crop rotational system were investigated over two-years. In this study, four cover crop treatments included 1) wheat (Triticum aestivum); 2) red clover (Trifolium pratense); 3) rapeseed (Brassica napus); and 4) fallow in the crop rotation. The amount of triple superphosphate as a P fertilizer applied rate after cultivation of cover crops was 120 and $360k\;ha^{-1}$ in 2014 and 2015, respectively. Soybean roots were sampled at full-flowering and analyzed for AMF communities using polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) and quantitative real-time PCR (qPCR) techniques. The AMF root colonization in the soybean roots at full bloom stage was significantly influenced by cover crop and P application throughout the two-year rotation. The two-year rotation of different cover crops or fallow impacted the molecular diversity of AMF communities colonizing roots of soybean. Redundancy analysis (RDA) indicated that AMF communities colonizing roots of soybean were significantly different among cover crop rotations. The AMF communities colonizing roots of soybean were clearly influenced by a P application in the two-year trial. Moreover, a P application may have positively impacts on the AMF communities under P-deficit soil due to the continuous cover crop-soybean rotational system without a P fertilizer.

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Comparative physiological and proteomic analysis of leaf in response to cadmium stress in sorghum

  • Roy, Swapan Kumar;Cho, Seong-Woo;Kwon, Soo Jeong;Kamal, Abu Hena Mostafa;Kim, Sang-Woo;Lee, Moon-Soon;Chung, Keun-Yook;Woo, Sun-Hee
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.124-124
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    • 2017
  • Cadmium (Cd) is of particular concern because of its widespread occurrence and high toxicity and may cause serious morpho-physiological and molecular abnormalities in in plants. The present study was performed to explore Cd-induced morpho-physiological alterations and their potentiality associated mechanisms in Sorghum bicolor leaves at the protein level. Ten-day-old sorghum seedlings were exposed to different concentrations (0, 100, and $150{\mu}M$) of $CdCl_2$, and different morpho-physiological responses were recorded. The effects of Cd exposure on protein expression patterns in S. bicolor were investigated using two-dimensional gel electrophoresis (2-DE) in samples derived from the leaves of both control and Cd-treated seedlings. The observed morphological changes revealed that the plants treated with Cd displayed dramatically altered shoot lengths, fresh weights, and relative water content. In addition, the concentration of Cd was markedly increased by treatment with Cd, and the amount of Cd taken up by the shoots was significantly and directly correlated with the applied level of Cd. Using the 2-DE method, a total of 33 differentially expressed protein spots were analyzed using MALDI-TOF/TOF MS. Of these, treatment with Cd resulted in significant increases in 15 proteins and decreases in 18 proteins. Significant changes were absorbed in the levels of proteins known to be involved in carbohydrate metabolism, transcriptional regulation, translation and stress responses. Proteomic results revealed that Cd stress had an inhibitory effect on carbon fixation, ATP production and the regulation of protein synthesis. In addition, the up-regulation of glutathione S-transferase and cytochrome P450 may play a significant role in Cd-related toxicity and stress responses. Our study provides insights into the integrated molecular mechanisms involved in response to Cd and the effects of Cd on the growth and physiological characteristics of sorghum seedlings. The upregulation of these stress-related genes may be candidates for further research and use in genetic manipulation of sorghum tolerance to Cd stress.

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Detection and Identification of Mycobacterium Tuberculosis in Patients with Tuberculous Cervical Lymphadenitis by PCR-RFLP (경부 결핵성 임파선염 환자에서 PCR-RELP를 이용한 결핵균의 검출 및 확인)

  • Lee Sang-Sook;Cho Young-Rok;Chun Ji-Min;Choi Yong-Seok;Sohn Eun-Ju;Park Nam-Cho;Park June-Sik
    • Korean Journal of Head & Neck Oncology
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    • v.12 no.2
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    • pp.169-176
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    • 1996
  • Tuberculous cervical lymphadenitis is still an important cause of neck mass in Korea. Tuberculosis is an important differential diagnosis in patients of cervical lymphadenopathy. Rapid and sensitive test for the diagnosis of tuberculosis is essential for the approapiate treatment. Up to now, conventional diagnostic methods for M. tuberculosis were acid-fast bacilli(AFB) stain and culture of M. tuberculosis. The direct microscopic examination of AFB by Ziehl-Neelsen stain is rapid, but often negative. The culture for M. tuberculosis is time-consuming, taking 4 to 8 weeks. Recently various methods to detect Mycobacterial DNA, including PCR and restriction fragment length polymorphism(RFLP) analysis have been reported. Here we represent a simple method for the confirmation of M. tuberculosis and exclusion of the other Mycobacterial species by RFLP analysis and silver staining of polyacrylamide gel electrophoresis after nested PCR for a repetitive DNA sequence(IS986) specific for M. tuberculosis from fresh or paraffin-embedded biopsy specimens. This result leads us to conclude that this method is simple, rapid and possibly applicable to confirm M. tuberculosis and rule out the other Mycobacteria species from the clinical specimens in the clinical laboratories.

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